Rehabilitation of contaminated soils and water using “tailor-made” Technosols

INTERNATIONAL DOCTORAL SCHOOL OF THE USC Maria João Cerveira Fernandes PhD Thesis Rehabilitation of contaminated soils and water using “tailormade” Technosols Santiago de Compostela, 2023 Doctoral Programme in Chemical and Environmental Engineering TESIS DE DOCTORADO REHABILITATION OF CONTAMINATED SOILS AND WATER USING “TAILORMADE” TECHNOSOLS Maria João Cerveira Fernandes ESCUELA DE DOCTORADO INTERNACIONAL DE LA UNIVERSIDAD DE SANTIAGO DE COMPOSTELA PROGRAMA DE DOCTORADO EN INGENIERÍA QUÍMICA Y AMBIENTAL SANTIAGO DE COMPOSTELA / LUGO 2023 ii DECLARACIÓN DO AUTOR/A DA TESE D./Dna. Maria João Cerveira Fernandes Título da tese: Rehabilitation of contaminated soils and water using “tailor-made” Technosols Presento a miña tese, seguindo o procedemento axeitado ao Regulamento, e declaro que: 1) A tese abarca os resultados da elaboración do meu traballo. 2) De ser o caso, na tese faise referencia ás colaboracións que tivo este traballo. 3) Confirmo que a tese non incorre en ningún tipo de plaxio doutros autores nin de traballos presentados por min para a obtención doutros títulos. 4) A tese é a versión definitiva presentada para a súa defensa e coincide a versión impresa coa presentada en formato electrónico E comprométome a presentar o Compromiso Documental de Supervisión no caso de que o orixinal non estea na Escola. En Santiago de Compostela, 03 de Abril de 2023. Sinatura electrónica AUTORIZACIÓN DEL TUTOR DE LA TESIS Rehabilitation of contaminated soils and water using “tailormade” Technosols D. Juan Manuel Lema Rodicio INFORMA: Que la presente tesis, se corresponde con el trabajo realizado por Dª. Maria João Cerveira Fernandes, bajo mi tutorización, y a utorizo su presentación , considerando que reúne os r equisitos exigidos en el R eglamento de Estudios de Doctorado de la USC, y que como director de esta no incurre en las causas de abstención establecidas en la Ley 40/2015. De acuerdo con lo indicado en el Reglamento de Estudios de Doctorado, declara también que la presente tesis doctoral es idónea para ser defendida en base a la modalidad Monográfica con reproducción de publicaciones, en los que la participación del doctorando/a fue decisiva para su elaboración y las publicaciones se ajustan al Plan de Investigación. En Santiago de Compostela, 30 de mayo de 2023 AUTORIZACIÓN DO DIRECTOR / TITOR DA TESE Rehabilitation of contaminated soils and water using “tailormade” Technosols D./Dna. Felipe Macias Vazquez D./Dna. Maria Cristina Fernandes Delerue Alvim de Matos INFORMA/N: Que a presente tese, correspóndese co traballo realizado por D/Dna. Maria João Cerveira Fernandes, baixo a miña dirección/titorización, e a utorizo a súa presentación , considerando que reúne os r equisitos esixidos no R egulamento de Estudos de Doutoramento da USC, e que como director desta non incorre nas causas de abstención establecidas na Lei 40/2015. De acordo co indicado no Regulamento de Estudos de Doutoramento, declara tamén que a presente tese de doutoramento é idónea para ser defendida en base á modalidade de Monográfica con reproducción de publicaciones , nos que a participación do/a doutorando/a foi decisiva para a súa elaboración e as publicacións se axustan ao Plan de Investigación. En Santiago de Compostela, 3 de Abril de 2023 Maria João Cerveira Fernandes iii ACKNOWLEDGMENTS I would like to express my sincere gratitude to Doctor Professor Cristina Delerue-Matos, my supervisor, for her invaluable guidance, support, and encouragement throughout my research journey. Her expertise and feedback were crucial in shaping this thesis, and I am deeply grateful for her mentorship. In addition, I would like to thank her for the care and concern she has shown for the personal side of this long, difficult but beautiful journey. I am also deeply grateful to Doctor Professor Felipe Macías Vásquez, also my supervisor, for his wide knowledge, insightful and constructive comments, suggestions, and feedback that have helped me refine my research and bring it to completion. Additionally, and in spite of all difficulties, I thank him for the kindness and friendly smile that he has always given me, in good and not so good moments. I am also grateful to Doctor Sonia Figueiredo, who have played a significant role in the development of my scientific papers, always with a good advice. I would also like to thank Doctor Manuela Carvalho for her kindness and friendship, along with her suggestions and feedback. They were precious to me in the development of this trip. I extend my appreciation to all my colleagues from GRAQ/REQUIMTE and IIT-LTA, for their valuable contributions to my research. Their help, experience, and friendship made my time in the lab enjoyable and rewarding. In particular, I would like to extend a heartfelt thanks to: Diana and Maria, for their invaluable help and support during every step of this journey, making it always lighter. Their friendship, advice, and encouragement have been a constant source of inspiration and motivation, as well as their professional and personal support. Paula, its invaluable help, fellowship, and support have made this challenging process much more manageable and enjoyable. Sara and Luz, for they friendship and support during this journey. I would like to thank my family and friends for their support, encouragement, and love throughout my academic journey. Their belief in me and my abilities has been a constant source of motivation and inspiration. Acknowledgments iv Finally, I would like to express my deep gratitude to my parents and brother for their unwavering support, love, and encouragement throughout all my life. They have always been a solid basis for me, and their support has been instrumental for me to achieve my goals and be where I am today. I am also grateful to my husband, Paulo, for his love, support, and everyday encouragement. His kindness, positivity, and capacity for transform from husband to father, and sometimes mother, have been a constant source of inspiration and motivation. I must also thank my son, Francisco, for coming to transform our lives into something much more meaningful and for teaching me new things every day. Maria João Cerveira Fernandes v RESUMO A contaminación ambiental, a alteración dos procesos ambientais naturais que afectan tanto aos compoñentes físicos como biolóxicos do sistema Terra/atmosfera, representa un problema grave e actual para a sociedade moderna. O notable crecemento da poboación provocou cambios profundos nos patróns de consumo, incluíndo as necesidades alimentarias e sanitarias, e o conseguinte uso excesivo de substancias como pesticidas (produción de alimentos) e farmacéuticos (saúde humana e animal). Tanto os produtos farmacéuticos como os pesticidas considéranse cada vez máis produtos químicos de preocupación emerxente, con efectos negativos coñecidos sobre as especies do solo e acuáticas e os seus respectivos ecosistemas, así como efectos potenciais sobre a saúde humana. A fluoxetina é un antidepresivo moi utilizado para tratar a depresión. Como ocorre con outros produtos farmacéuticos, só unha parte deste composto é metabolizada polo corpo humano, mentres que o resto, así como os seus metabolitos, son excretados, chegando finalmente aos sistemas de augas residuais e potencialmente ao medio ambiente. O glifosato e o glufosinato, dous herbicidas organofosforados non selectivos de amplo espectro, e o ácido aminometilfosfónico (AMPA), o principal metabolito do glifosato, tamén son tres compostos de interese. O nivel de exposición de humanos e animais e os efectos nocivos destes compostos están amplamente documentados. Ademais doutros contaminantes emerxentes, a contaminación ambiental por fluoxetina, glifosato, glufosinato e AMPA require de medidas correctoras, impoñendo o desenvolvemento de tecnoloxías de remediación para a rehabilitación das zonas afectadas. Estudáronse e desenvolvéronse diversas tecnoloxías de remediación para facer fronte ás contaminacións ambientais. Unha destas tecnoloxías de remediación é a adsorción, un método no que as moléculas ou partículas dunha substancia son retidas na superficie dun material sólido ou líquido a través de diversos mecanismos, como a atracción electrostática, forzas de Van der Waals ou enlaces químicos. Úsase habitualmente en procesos industriais para eliminar impurezas de líquidos ou gases, así como na rehabilitación de solos, tratamento de augas residuais e depuración de auga potable. Ademais, é un proceso que se produce de forma espontánea en diversos sistemas naturais, como no solo, beneficiando a absorción de nutrientes por parte das plantas ou a adsorción de gases da atmosfera polas partículas do solo. A adsorción inflúe significativamente na transferencia e dispoñibilidade de pesticidas no solo, sendo un dos principais procesos para controlar a lixiviación dos pesticidas e reducir a Resumo xii de óxidos de aluminio e ferro, arxila e materia orgánica. O desenvolvemento de Tecnosoles “a la carta” ofrece unha solución innovadora e integrada que contribúe a mellorar a calidade dos recursos naturais, promovendo a sustentabilidade, reducindo a produción de residuos e restaurando zonas contaminadas. En xeral, os coñecementos adquiridos permiten coñecer mellor o estado actual de contaminación ambiental por produtos farmacéuticos e praguicidas, xa que demostra que a contaminación non só chega aos cursos de auga, senón que tamén afecta aos sedimentos e ás augas subterráneas. Esta investigación proporciona datos valiosos sobre a adsorción de fluoxetina, glifosato, AMPA e glufosinato en diferentes materiais. Os estudos de adsorción mostraron, entre outros resultados, que os materiais biochar probados xurdiron como excelentes materiais adsorbentes de fluoxetina pero, pola contra, non teñen unha boa capacidade de retención de compostos como o glifosato, o AMPA e, especialmente, o glufosinato. Por outra banda, os solos e Tecnosoles mostraron unha alta capacidade de retención de glifosato e AMPA. O glufosinato non tendeu a adsorbirse a ningún dos materiais ensaiados. Os resultados apoian o desenvolvemento de tecnoloxías de remediación destinadas a mitigar o impacto ambiental destes compostos, previr a contaminación e salvagardar a saúde pública. Ademais, propoñen o uso do biochar como compoñente dos tecnosoles producidos para reter a fluoxetina. Tamén destaca algunhas das características fundamentais que se requiren dos constituíntes do tecnosol para a correcta adsorción do glifosato e do AMPA. Maria João Cerveira Fernandes xiii RESUMEN La contaminación ambiental, la alteración de los procesos ambientales naturales que afectan tanto a los componentes físicos como biológicos del sistema Tierra/atmósfera, representa un problema grave y actual para la sociedad moderna. El notable crecimiento demográfico ha provocado profundas alteraciones en los patrones de consumo, incluyendo las necesidades alimentarias y sanitarias, y el consiguiente uso excesivo de sustancias como los plaguicidas (producción de alimentos) y los productos farmacéuticos (salud humana y animal). Tanto los productos farmacéuticos como los plaguicidas se consideran cada vez más sustancias químicas de preocupación emergente, con efectos negativos conocidos sobre el suelo y las especies acuáticas y sus respectivos ecosistemas, así como efectos potenciales sobre la salud humana. La fluoxetina es un antidepresivo ampliamente utilizado para tratar la depresión. Como ocurre con otros productos farmacéuticos, sólo una parte de este compuesto es metabolizada por el cuerpo humano, mientras que el resto, así como sus metabolitos, son excretados, alcanzando eventualmente los sistemas de aguas residuales y potencialmente el medio ambiente. El glifosato y el glufosinato, dos herbicidas organofosforados no selectivos de amplio espectro, y el ácido aminometilfosfónico (AMPA), metabolito principal del glifosato, son también tres compuestos de interés. El nivel de exposición de humanos y animales y los efectos nocivos de estos compuestos están ampliamente documentados. Así como otros contaminantes emergentes, la contaminación ambiental por fluoxetina, glifosato, glufosinato y AMPA, requiere medidas correctoras, imponiéndose el desarrollo de tecnologías de remediación para la rehabilitación de las zonas afectadas. Se han estudiado y desarrollado varias tecnologías de remediación para tratar las contaminaciones medioambientales. Una de estas tecnologías de remediación es la adsorción, un método en el que las moléculas o partículas de una sustancia son retenidas en la superficie de un material sólido o líquido a través de distintos mecanismos, como la atracción electrostática, las fuerzas de Van der Waals o los enlaces químicos. Se utiliza habitualmente en procesos industriales para eliminar impurezas de líquidos o gases, así como en la rehabilitación de suelos, en el tratamiento de aguas residuales y en la purificación de agua potable. Además, es un proceso que ocurre espontáneamente en diversos sistemas naturales, como en el suelo, beneficiando la absorción de nutrientes por las plantas o la adsorción de gases de la atmósfera por las partículas del suelo. La adsorción influye significativamente en Resumen xiv la transferencia y disponibilidad de los plaguicidas en el suelo, siendo uno de los principales procesos de control de la lixiviación de plaguicidas y reduciendo su migración y el potencial de contaminación de las aguas superficiales o subterráneas. Dado que se utiliza ampliamente en tratamientos de aguas residuales a larga escala, debido a su sencilla estructura, su alta eficiencia y la posibilidad de utilizar adsorbentes de bajo coste y no tóxicos, puede optimizarse y aplicarse a la eliminación de productos farmacéuticos y plaguicidas. Para estudiar la adsorción de diferentes tipos de sustancias como la fluoxetina, el glifosato, el AMPA y el glufosinato, es esencial evaluar el uso de una amplia variedad de materiales adsorbentes y sus propiedades fisicoquímicas. Su selección es de gran importancia, y características como el bajo coste, la sostenibilidad, la no toxicidad y la alta capacidad de sorción son atributos significativos a tener en cuenta. El biochar es un material poroso sólido y estable con alto contenido en carbono, producido a través de la pirólisis/carbonización de biomasa en un entorno de oxígeno limitado o ausente y en condiciones de temperatura moderada. Es un material atractivo para ser implementado como adsorbente para retener y eliminar contaminantes de ambientes sólidos y acuosos. Su uso se presenta como una herramienta para la gestión medioambiental, actuando en la mejora de la calidad de los suelos, en el tratamiento de residuos, en la mitigación del cambio climático y en la producción de energía. Además de ser económicamente atractivos, los biochar se utilizan en gran medida como adsorbentes debido a su alta capacidad de retención de distintos contaminantes asociada a su elevada superficie específica y porosidad, capacidad de intercambio iónico y pH. La selección de la materia prima puede influenciar ampliamente sus propiedades, así como las condiciones del proceso de producción, es decir, la temperatura, la rapidez de calentamiento, la presión y el tiempo de residencia. Otra tecnología innovadora de remediación es el uso de Tecnosoles “a la carta”. Los Tecnosoles son un grupo de suelos de referencia definido en la World Reference Base para los recursos edáficos (de la Organización de las Naciones Unidas para la Agricultura y la Alimentación y la Unión Internacional de Ciencias del Suelo) como una combinación de suelos "cuyas propiedades y pedogénesis están dominadas por su origen técnico". Estos suelos, dominados o fuertemente afectados por materiales fabricados por el hombre y fuertemente modificados (denominados artefactos, como residuos de materiales como ladrillos, vidrio, residuos domésticos, residuos de minas, lodos, entre otros) o extraídos de grandes profundidades, se encuentran con frecuencia en zonas urbanas e industriales. Los Tecnosoles "a la carta" son suelos artificiales creados mediante la combinación de suelo natural con Maria João Cerveira Fernandes xv materiales antropogénicos, derivados de actividades humanas, con otros aditivos para obtener las características deseadas. Replican las funciones del suelo natural y pueden utilizarse para proporcionar varias funciones del suelo y servicios ecosistémicos, como condiciones adecuadas para el crecimiento/producción de plantas (alimentos y biomasa), la recuperación del suelo, el secuestro de carbono, la restauración de masas de agua y zonas degradadas o contaminadas, la reposición de aguas subterráneas y la biodiversidad de los ecosistemas. El biochar es uno de los materiales que pueden incorporarse en la producción de Tecnosoles "a la carta". Así, como una primera aproximación a los problemas abordados en este trabajo, se llevó a cabo el monitoreo de antibióticos y antidepresivos en las aguas y sedimentos de dos ríos del norte de Portugal. Se recogieron muestras de sedimentos de dos camadas, la camada superior, entre 0 y 2 cm de profundidad, y la camada inferior, entre 2 y 10 cm. El presente estudio incluyó la caracterización y el análisis de muestras para diecisiete antibióticos y diez antidepresivos. La comparación de las concentraciones de fármacos en las muestras del río Leça y del río Douro reveló niveles más elevados en el río Leça. Sólo se detectaron carbamazepina y fluoxetina en las muestras de agua del Douro, mientras que las muestras de agua de Leça contenían trece de los veintisiete productos farmacéuticos, incluidos seis antibióticos y siete fármacos psiquiátricos. En particular, la azitromicina se encontró en la concentración más alta (2.82 µg/L) en las muestras del río Leça, especialmente cerca de un punto de descarga de efluentes de una planta de tratamiento de aguas residuales. La fluoxetina presentó la mayor frecuencia de detección en las muestras de agua. La presencia de carbamazepina y venlafaxina en el agua del río Leça fue reveladora de riesgos potenciales para las algas. En las muestras de sedimentos, se encontró sulfametoxipiridazina en el sedimento del Douro, mientras que se detectó azitromicina en el sedimento de Leça, ambas principalmente en la camada superior del sedimento. También se detectaron fármacos psiquiátricos, cuatro en el sedimento de Douro y seis en el de Leça. Una comparación entre las camadas superior e inferior del sedimento en cada punto de muestreo mostró que la venlafaxina, el único fármaco cuantificado en el río Douro, presentaba una mayor concentración en la zona inferior, potencialmente relacionada con la migración del suelo. En los sedimentos de Leça, se observaron concentraciones más elevadas en la camada superior, probablemente asociadas a episodios recientes de contaminación en la zona. Una revisión de los estudios de monitorización reportados en la literatura reveló una inclusión reducida de análisis de sedimentos, centrándose la mayoría de los estudios únicamente en muestras de agua (especialmente para drogas psiquiátricas). Por lo tanto, no fue posible identificar un patrón de contaminación consistente ni establecer un vínculo claro entre la contaminación de Resumen xvi los sedimentos y la migración de contaminantes del agua, basándose en los resultados obtenidos y en las comparaciones bibliográficas. Sin embargo, se observaron sistemáticamente concentraciones más elevadas de contaminantes en los lugares de muestreo situados aguas abajo de los vertidos de las plantas de tratamiento de aguas residuales. Además, se realizó una revisión bibliográfica de la contaminación del agua y los sedimentos por glifosato, AMPA y glufosinato en todo el mundo. El objetivo fue investigar la contaminación y la movilidad de las sustancias referidas en los medios acuáticos, con especial atención a las muestras de agua y sedimentos. La investigación incluyó una exhaustiva revisión bibliográfica para analizar los estudios publicados en los últimos cinco años que monitorizaron las concentraciones de estos compuestos en agua (tanto superficial como subterránea) y sedimentos. El análisis de la legislación aplicable reveló variaciones significativas en los valores límite de estos contaminantes en los distintos países. Por ejemplo, la Unión Europea estableció el límite legal más bajo para los residuos de glifosato en el agua destinada al consumo humano, con un límite de 0.1 μg/L, mientras que Japón estableció el límite más alto en 2 000 μg/L. Los métodos de detección más utilizados en los estudios analizados fueron la cromatografía líquida de alta resolución (HPLC) en tándem con la espectrometría de masas (MS/MS). Sin embargo, se encontró que sólo el 40% de los estudios revisados reportaron límites de detección por debajo del valor máximo permisible de la Unión Europea para la contaminación del agua por plaguicidas individuales (0.1 μg/L). La investigación del monitoreo del glifosato en el agua y los sedimentos reveló tendencias interesantes. Francia resultó ser el país con la mayor relación muestras/publicaciones, seguido de Japón e Italia. Argentina, por su parte, presentó una mayor frecuencia de publicaciones de estudios, pero una menor relación muestras/publicaciones. En particular, los países europeos que realizaron un mayor número de análisis del nivel de glifosato tendieron a corresponderse con una mayor tasa de aplicación de glifosato. En cuanto al seguimiento de AMPA, Argentina presentó una mayor frecuencia de estudios, mientras que Francia mantuvo su posición como país con la mayor relación muestras/publicación, seguida de Estados Unidos, Suecia e Italia. China se destacó con un número significativamente mayor de muestras monitoreadas para glufosinato en comparación con otros países con publicaciones sobre este compuesto. Las concentraciones máximas de contaminantes variaron en función de la matriz ambiental. En aguas superficiales, las concentraciones más altas se encontraron para el AMPA (6 872 μg/L), seguido del glifosato (5 750 μg/L) y el glufosinato (13.15 μg/L). En las aguas subterráneas, los valores máximos fueron de 8 700 μg/L para el glifosato, 233 μg/L para el AMPA y 0.03 μg/L para el glufosinato. Los sedimentos mostraron las mayores concentraciones detectadas de glufosinato (14.9 Maria João Cerveira Fernandes xvii μg/kg), mientras que el glifosato y el AMPA alcanzaron concentraciones máximas de 1 882 μg/kg y 4 033 μg/kg, respectivamente. Contrariamente a lo esperado, la concentración máxima de glifosato fue mayor en las aguas subterráneas que en las superficiales. Sin embargo, la frecuencia de detección fue significativamente mayor en las aguas superficiales (70%) en comparación con las subterráneas (18%). En cuanto al AMPA y al glufosinato, la frecuencia de detección en las aguas superficiales fue del 87% y del 6%, respectivamente, mientras que en las aguas subterráneas fue del 16% y del 0.7%. Los países sudamericanos presentaron los valores más altos de concentración máxima detectada para el glifosato y el AMPA, mientras que los sedimentos fueron la matriz ambiental con mayor concentración de glufosinato. Francia presentó las mayores concentraciones máximas de glifosato en aguas superficiales, seguida de Italia y Alemania. En las aguas subterráneas, Suecia presentó las concentraciones máximas de glifosato, seguida de Italia. Alemania y Croacia presentaron los niveles máximos de concentración en sedimentos. Se identificaron varios factores como impulsores de la dispersión de estos compuestos en aguas superficiales, sedimentos y aguas subterráneas. Estos factores incluían el exceso de pulverización, la deriva de la pulverización, las escorrentías superficiales, los niveles freáticos poco profundos, la baja conductividad hidráulica de los acuíferos, el bajo gradiente hidráulico, la velocidad muy baja del flujo, el tipo de suelo y la constitución química. La aplicación persistente y creciente de estos compuestos en actividades agrícolas y domésticas, el uso inadecuado y las plantas de tratamiento de aguas residuales se identificaron como fuentes potenciales de contaminación. Esta revisión aclaró el problema de la contaminación y la movilidad del glifosato, el AMPA y el glufosinato en el agua y los sedimentos. Los resultados subrayaron la necesidad de llevar a cabo un seguimiento continuo y de desarrollar estrategias de remediación para mitigar el impacto ambiental de estos compuestos. Los resultados también pusieron en relieve la importancia de una legislación armonizada y unos valores límite unificados en todos los países para garantizar una protección eficaz de los recursos hídricos. A la vista de la contaminación documentada asociada a los compuestos investigados, es crucial identificar tecnologías de remediación que no sólo se ocupen de los lugares contaminados, sino que también reduzcan la movilidad de estos compuestos. Estas tecnologías pueden evitar la contaminación del agua y del suelo y minimizar los problemas de salud asociados a la ingestión de estos compuestos. En consecuencia, esta investigación se centró en el estudio de la adsorción de fluoxetina, glifosato, AMPA y glufosinato sobre diversos materiales, con diferentes características, con el fin de comprender y optimizar su adsorción. Con este fin, se desarrollaron estudios de adsorción, a saber, ensayos de equilibrio, cinéticos y de influencia del pH, para estudiar la adsorción de los cuatro compuestos Resumen xviii analizados. Se ensayaron diferentes adsorbentes con características distintas para encontrar los materiales y características más adecuados para integrar la composición de un Technosol "a la carta". Si se aplican a la producción de un Technosol "a la carta”, los resultados permitirán desarrollar una solución innovadora e integrada que contribuya a mejorar la calidad de los recursos naturales, la sostenibilidad, la reducción de la producción de residuos y la restauración de zonas contaminadas. En la primera fase del estudio se ensayaron doce biochars derivados de diferentes residuos forestales y agroalimentarios (podas de Quercus ilex, Eucalyptus grandis, Pinus pinaster, Quercus suber, Malus pumila, Prunus spinosa, Cydonia oblonga, Eriobotrya japonica, Juglans regia, Actinidia deliciosa, Citrus sinensis y Vitis vinifera) como adsorbentes potenciales, de bajo coste y renovables, para la eliminación de la fluoxetina del agua. Los experimentos preliminares de adsorción permitieron seleccionar los adsorbentes más prometedores, Quercus ilex (encina), Cydonia oblonga (membrillo), Eucalyptus, Juglans regia (nogal) y material de poda de Vitis vinifera (vid). Se caracterizaron mediante análisis proximal, elemental y mineral, análisis termogravimétrico, espectroscopia infrarroja por transformada de Fourier, determinación de la superficie específica y pH en el punto de carga cero (pHPZC). Se realizaron estudios por batch y de equilibrio, y se evaluó la influencia del pH. Se determinaron las capacidades de adsorción de los biochars seleccionados, con valores que oscilaban entre 2.21 y 6.41 mg/g según el modelo de Langmuir. Entre los biochars ensayados, el biochar de eucalipto mostró la mayor capacidad de adsorción, seguido de los biochars de encina, vid, nogal y membrillo. La cinética de adsorción fue rápida, produciéndose la adsorción completa en menos de 15 minutos. Los experimentos de adsorción en columna realizados con biochar de eucalipto demostraron su eficacia en modo continuo, presentando un avance del 50% de eliminación a los 90 minutos, lo que sugiere su potencial como alternativa renovable para la eliminación de fluoxetina. La siguiente fase de la investigación se centró en investigar la adsorción de glifosato, AMPA y glufosinato en cuarenta y un materiales diferentes, incluidos biochars, suelos y Tecnosoles "a la carta" (TMT). Los biochars ensayados se produjeron a partir de troncos de acacia (Acacia melanoxylon) divididos en albura (BAcSw-3) y duramen (BAcHw-2), ramas de eucalipto (BEu-2), serrín de pino (BSd-1), corteza de pino (BPb), compost de orujo de oliva (BOpc), cascarilla de arroz (BRh), mazorca de maíz (BCc1 y BCc-2), y astillas de madera trituradas alemanas (BSdG). Las ocho muestras de suelo analizadas eran Umbrisoles alumínicos (S4A, 11B, 13A), Ferrasoles derivados de anfibolitas (S9B), Umbrisoles Háplicos (1Ah y 2Ah) y Leptosoles Ándicos y Ferrálicos. En relación con el TMT, los materiales ensayados Maria João Cerveira Fernandes xix se clasificaron como Tecnosoles arénicos cálcicos (TMT 1), Tecnosoles aluándicos (TMT 2), Tecnosoles ándicos eútricos (TMT 3, G, I, J, K, L, M, Q, R y S), Tecnosoles eútrico reductores (TMT 4 y A), Tecnosoles dístricos (TMT B y D), Tecnosoles cálcicos (TMT C), Tecnosoles ándicos (TMT E y F), Tecnosoles eútricos (TMT H y O), Tecnosol férrico (TMT N) y Tecnosoles ándicos reductores (TMT P). Estos TMT se produjeron a partir de residuos industriales y agroindustriales, a saber, sedimentos de mina, biomasa de serrín, lodos del tratamiento de aguas residuales, compost orgánico, cenizas y arena. Los resultados indicaron que los tres compuestos fueron pobremente adsorbidos por la mayoría de las muestras de biochar, con tasas máximas de eliminación del 25% para el glifosato, 12% para el AMPA, y 4% para el glufosinato. Estos resultados se atribuyeron principalmente a la temperatura de producción de los biochar. Por el contrario, los suelos ensayados mostraron altas capacidades de adsorción para glifosato y AMPA, con tasas máximas de eliminación del 94% y 91% en los ensayos preliminares, respectivamente. Sin embargo, la adsorción de glufosinato fue limitada, con tasas de eliminación que no superaron el 6%. Los TMT también mostraron capacidades de adsorción significativas durante los ensayos preliminares, con tasas de eliminación máximas del 79% para el glifosato y del 83% para el AMPA. Una vez más, el glufosinato mostró valores de adsorción bajos, con una tasa de eliminación máxima de sólo el 3%. Además, se observó que los tamaños de grano más pequeños mejoraban el rendimiento de la adsorción en comparación con los suelos o TMT con tamaños de grano más grandes. El tiempo de equilibrio para la adsorción varió, produciéndose la adsorción total en 14 horas en el caso de los suelos y en 72 horas en el de los TMT, lo que concuerda con hallazgos anteriores en la bibliografía. Las capacidades máximas de adsorción, según el modelo de Langmuir, se determinaron en 10.84 mg/g para el glifosato (suelo S9B) y 15.08 mg/g para el AMPA (TMT 1). En cuanto a la influencia del pH en el proceso de adsorción, el glifosato y el ácido aminometilfosfónico (AMPA) presentan comportamientos de carga distintos. Analizando el impacto del pH en el proceso de adsorción con los adsorbentes ensayados, que tienen un pHPZC de aproximadamente 4.9, la adsorción de glifosato y AMPA exhibió un comportamiento dependiente del pH. La adsorción aumentó entre pH 1 y pH 3, correspondiendo a la pérdida de protones asociada al grupo carboxilo del glifosato (pKa2) y al grupo fosfónico del AMPA (pKa1) en presencia de una superficie adsorbente predominantemente cargada positivamente. Posteriormente, se produjo una disminución gradual de la adsorción entre pH 3 y pH 6, más pronunciada para el glifosato que para el AMPA. Entre pH 6 y 10, la adsorción de glifosato en los adsorbentes ensayados disminuyó significativamente, excepto en el caso de TMT 1, que sólo mostró una adsorción reducida a pH superiores a 10, acercándose a valores próximos a cero. En el caso del AMPA, la reducción Resumen xx de la adsorción por los TMT se produjo de forma más destacada a partir de valores de pH superiores a 9, observándose una adsorción relativamente constante entre pH 6 y 10 para el resto de adsorbentes. Para todos los adsorbentes, se produjo una notable disminución de la adsorción hasta valores cercanos a cero entre pH 10 y 12, excepto para la adsorción de glifosato en el suelo S4A, que mostró una adsorción ligeramente superior a pH 12 en comparación con los otros adsorbentes. En general, la mayor adsorción de glifosato y AMPA en suelos y TMTs ocurrió dentro del rango de pH de 2 a 5, con las superficies de los adsorbentes predominantemente cargadas positivamente, y aunque la principal forma iónica del AMPA dentro de este rango de pH es neutra. Estos hallazgos demuestran que los suelos naturales y los TMT pueden servir como adsorbentes eficaces, de bajo coste y sostenibles para eliminar el glifosato y el AMPA de soluciones acuosas. Además, para la producción de TMT deberían seleccionarse materiales con pH relativamente bajo y alto contenido en óxidos de aluminio y hierro, arcilla y materia orgánica. El desarrollo de Tecnosoles a medida ofrece una solución innovadora e integrada que contribuye a mejorar la calidad de los recursos naturales, promover la sostenibilidad, reducir la producción de residuos y restaurar las zonas contaminadas. En general, los conocimientos adquiridos permiten comprender mejor el estado actual de la contaminación ambiental por productos farmacéuticos y plaguicidas, pues demuestran que la contaminación no sólo llega a los cursos de agua, sino que también afecta a los sedimentos y las aguas subterráneas. Esta investigación aporta valiosos datos sobre la adsorción de fluoxetina, glifosato, AMPA y glufosinato en distintos materiales. Los estudios de adsorción mostraron, entre otros resultados, que los materiales de biochar ensayados se erigieron como excelentes materiales adsorbentes de fluoxetina pero, por el contrario, no presentan una buena capacidad de retención a compuestos como glifosato, AMPA y, especialmente, glufosinato. Por otro lado, los suelos y los Technosols han mostrado una alta capacidad de retención para el glifosato y el AMPA. El glufosinato no tendió a adsorberse en ninguno de los materiales ensayados. Los resultados apoyan el desarrollo de tecnologías de remediación destinadas a mitigar el impacto ambiental de estos compuestos, prevenir la contaminación y salvaguardar la salud pública. Además, proponen el uso de biochar como componente de Tecnosoles producidos para retener la fluoxetina. También destaca algunas de las características clave que se requieren de los constituyentes del Tecnosol para una adsorción adecuada del glifosato y el AMPA. Maria João Cerveira Fernandes xxi ABSTRACT Environmental pollution, the disturbance of natural environmental processes that affect both the physical and biological components of the earth/atmosphere system, represents a serious and current problem for modern society. The substantial population growth has led to remarkable alterations in consumption patterns, including food and health necessities and consequent overuse of substances such as pesticides (food production) and pharmaceuticals (human and animal health). Both pharmaceuticals and pesticides are increasingly acknowledged as chemicals of emerging concern, with known negative impacts on soil and aquatic species and respective ecosystems and potential effects on human health. Fluoxetine is an anti-depressant pharmaceutical widely used to treat depression. As with other pharmaceuticals, only a part of this compound is metabolized by the human body, while the rest, as well as its metabolites, are excreted, eventually reaching the wastewater systems and potentially the environment. Glyphosate and glufosinate, two broad-spectrum, non-selective, organophosphorus herbicides, and aminomethylphosphonic acid (AMPA), the primary metabolite of glyphosate, are also three compounds of concern. The exposure level of humans and animals and the harmful effects of these compounds is largely documented. As well as other emerging pollutants, environmental contamination by fluoxetine, glyphosate, glufosinate, and AMPA, require corrective measures, imposing the development of remediation techniques for the rehabilitation of the affected areas. Several remediation technologies have been studied and developed to treat environmental contaminations. One of these remediation technologies is adsorption, a method in which molecules or particles of a substance are retained on the surface of a solid or liquid material through various mechanisms, such as electrostatic attraction, van der Waals forces, or chemical bonding. It is commonly used in industrial processes to remove impurities from liquids or gases, as well as in soil remediation, wastewater treatment, and drinking water purification. Also, it is a process that spontaneously occurs in many natural systems, such as in soil, benefiting the uptake of nutrients by plants or the adsorption of gases in the atmosphere by soil particles. Adsorption significantly affects the transfer and availability of pesticides in soil, being one of the main processes of controlling pesticide leaching and reducing their migration and the potential to contaminate surface waters or groundwater. As it is extensively used in large-scale wastewater treatments, due to its simple layout, high Abstract xxviii to retain fluoxetine. It also highlights some of the key characteristics required from the Technosol constituents for adequate adsorption of glyphosate and AMPA. Maria João Cerveira Fernandes xxix INDICE LIST OF ABBREVIATIONS ........................................................................................... XXXIII CHAPTER 1 INTRODUCTION .......................................................................................... 1 1.1 General background .................................................................................................................... 1 1.2 Objectives and structure ............................................................................................................. 2 1.2.1 Main objective ............................................................................................................................... 2 1.2.2 Specific objectives ......................................................................................................................... 3 1.2.3 Thesis outline ................................................................................................................................ 4 CHAPTER 2 LITERATURE REVIEW ................................................................................... 7 2.1 Environmental Pollution ............................................................................................................. 7 2.2 Pharmaceutical products ............................................................................................................ 9 2.2.1 Fluoxetine antidepressant use .................................................................................................... 16 2.2.2 Antidepressant Fluoxetine - chemical and physical properties .................................................. 18 2.2.3 Antidepressant Fluoxetine - degradation and dissipation paths ................................................ 20 2.2.4 Analytical methodologies for Fluoxetine detection in environmental matrixes ......................... 21 2.3 Pesticides environmental contamination ...................................................................................22 2.3.1 The use of glyphosate and glufosinate‐based herbicides ........................................................... 28 2.3.2 Chemical and physical properties of glyphosate and glufosinate ............................................... 33 2.3.3 Glyphosate and Glufosinate - degradation and dissipation paths .............................................. 35 2.3.4 Analytical methodologies for Glyphosate, AMPA and Glufosinate detection in environmental matrixes 40 2.4 Innovative technologies for mitigation of new contaminants ....................................................43 2.4.1 Adsorption ................................................................................................................................... 48 2.4.2 Prospective use of biochar to remediate contaminated sites .................................................... 49 2.4.3 Prospective use of tailor-made Technosols to remediate contaminated sites ........................... 52 2.4.4 Selection of the remediation technologies for glyphosate, AMPA, glufosinate, and fluoxetine contamination in soil and water ....................................................................................................................... 55 Indice xxx CHAPTER 3 ANTIBIOTICS AND ANTIDEPRESSANTS OCCURRENCE IN SURFACE WATERS AND SEDIMENTS COLLECTED IN THE NORTH OF PORTUGAL ................................................. 57 3.1 Introduction ...............................................................................................................................59 3.2 Materials and methods ..............................................................................................................61 3.2.1 Physical characteristics and points of pollution of the Douro and Leça rivers............................ 61 3.2.2 River waters and sediments sampling ......................................................................................... 62 3.2.3 River waters and sediments characterization ............................................................................. 63 3.2.4 Reagents and chemicals .............................................................................................................. 64 3.2.5 SPE cartridges and QuEChERS tube ............................................................................................. 65 3.2.6 River waters and sediments extraction procedures ................................................................... 65 3.2.7 UHPLC-MS/MS analysis ............................................................................................................... 65 3.2.8 Validation .................................................................................................................................... 66 3.2.9 Statistical analysis ....................................................................................................................... 66 3.2.10 Risk assessment ...................................................................................................................... 66 3.3 Results and discussion ...............................................................................................................66 3.3.1 Parameters .................................................................................................................................. 66 3.3.2 Method performance for pharmaceutical monitoring ............................................................... 68 3.3.3 River waters and sediments monitoring ..................................................................................... 72 3.3.4 Statistical analysis ....................................................................................................................... 76 3.3.5 Concentration found in literature ............................................................................................... 78 3.4 Conclusions ................................................................................................................................80 CHAPTER 4 GLYPHOSATE, AMPA AND GLUFOSINATE OCCURRENCE IN WATERS AND SEDIMENTS – WORLD ASSESSMENT ................................................................................... 111 4.1 Limits and threshold for Glyphosate, AMPA, and Glufosinate in water .................................... 112 4.2 Pre-treatment, extraction, and analysis ................................................................................... 115 4.3 Environmental monitoring ....................................................................................................... 118 4.4 Glyphosate, AMPA and glufosinate in water and sediments .................................................... 122 4.5 Presence of glyphosate, AMPA and glufosinate in other matrixes ........................................... 128 Maria João Cerveira Fernandes xxxi 4.6 Study and analysis of higher mobility pathways ...................................................................... 129 4.7 Glyphosate, AMPA and glufosinate contamination sources ..................................................... 132 CHAPTER 5 EVALUATION OF THE ADSORPTION POTENTIAL OF BIOCHARS PREPARED FROM FOREST AND AGRI-FOOD WASTES FOR THE REMOVAL OF FLUOXETINE .................... 135 Abstract ................................................................................................................................................ 137 5.1 Introduction ............................................................................................................................. 137 5.2 Materials and methods ............................................................................................................ 139 5.2.1 Chemicals .................................................................................................................................. 139 5.2.2 Biochar preparation .................................................................................................................. 139 5.2.3 Biochar characterization ........................................................................................................... 140 5.2.4 Preliminary adsorption assays................................................................................................... 141 5.2.5 Kinetic assays ............................................................................................................................ 142 5.2.6 Effect of pH ................................................................................................................................ 142 5.2.7 Equilibrium assays ..................................................................................................................... 143 5.2.8 Column assays ........................................................................................................................... 144 5.2.9 Determination of fluoxetine concentration .............................................................................. 145 5.3 Results and discussion ............................................................................................................. 145 5.3.1 Preliminary adsorption assays................................................................................................... 145 5.3.2 Biochar characterization ........................................................................................................... 146 5.3.3 Kinetic assays ............................................................................................................................ 148 5.3.4 pH studies .................................................................................................................................. 150 5.3.5 Adsorption isotherms ................................................................................................................ 152 5.3.6 Column adsorption assay .......................................................................................................... 154 5.4 Conclusions .............................................................................................................................. 155 CHAPTER 6 INFLUENCE OF ADSORBENT MATERIAL AND ITS CHARACTERISTICS ON THE GLYPHOSATE, AMPA AND GLUFOSINATE REMOVAL ............................................................ 161 Abstract ................................................................................................................................................ 163 Indice xxxii 6.1 Introduction ............................................................................................................................. 163 6.2 Materials and methods ............................................................................................................ 168 6.2.1 Chemicals and reagents ............................................................................................................ 168 6.2.2 Adsorbent Materials Preparation/Source ................................................................................. 168 6.2.3 Adsorbent Materials Characterization ...................................................................................... 169 6.2.4 Batch adsorption assays ............................................................................................................ 170 6.2.5 pH influence .............................................................................................................................. 172 6.2.6 Determination of glyphosate, AMPA, and glufosinate .............................................................. 172 6.3 Results and discussion ............................................................................................................. 173 6.3.1 Adsorbent Materials Characterization ...................................................................................... 173 6.3.2 Batch adsorption assays ............................................................................................................ 175 6.3.3 pH influence .............................................................................................................................. 186 6.3.4 Discussion .................................................................................................................................. 190 6.4 Conclusions .............................................................................................................................. 191 CHAPTER 7 GENERAL CONCLUSIONS ......................................................................... 201 7.1 Main outcomes of the thesis .................................................................................................... 201 7.2 Future research recommendations .......................................................................................... 205 REFERENCES ................................................................................................................ 207 LIST OF PUBLICATIONS ................................................................................................ 250 ANNEX - AUTHORIZATION FOR USE AND ADAPTATION OF FIGURES ............................. 255 List of Abbreviations xxxiii LIST OF ABBREVIATIONS AA Aceticacidanhydride ACN Acetonitrile AOP Advanced oxidation process EC Electrical conductivity CEC Chemicals of emerging concern CE-MS Capillary electrophoresis-mass spectrometry DAD Diode array detector EBS effective base saturation ECD Electrochemical detection EFSA European Food Safety Authority ELISA Enzyme-linked immunosorbent assay ESI-MS/MS Electrospray ionisation tandem mass spectrometry FAO Food and Agriculture Organization of the United Nations FLD Fluorescence detection FLX Fluoxetine FMOC-Cl 9-fluorenylmethylchloroformate GBH Glyphosate based herbicides GC Gas chromatography HFB Heptafluorobutanol HPAEC High-performance anion-exchange chromatography IC Ion chromatography with suppressed conductivity detection IC-HESI-MS/MS Ion chromatography hyphenated to electrospray tandem mass spectrometry ICP-MS/MS Inductively coupled plasma tandem mass spectrometry LC Liquid Chromatography List of Abbreviations xxxiv LOD Limit of detection LOQ Limit of quantification MDL Method detection limit MKP, KH2PO4 Monopotassium phosphate MQL Method quantification limit MS Mass spectrometry NSAIDs Non-Steroidal Anti-Inflammatory Drugs NBD-Cl 4-chloro-7-nitrobenzofurazan NPD Nitrogen-phosphorous detection OM Organic matter OPA-MCE o-phthalaldehyde-2-mercaptoethanol PAD Pulsed amperometric detection PFAS Per-fluoroalkyl and polyfluoroalkyl substances PTFE Polytetrafluoroethylene Q-TOF-MS Quadrupole Time-of-Flight Mass Spectrometry RP-HPLC Reverse phase high-performance liquid chromatography SIC Sequential injection chromatography SIM Selected ion monitoring SM-FIA Spectrophotometric method - standard flow injection analysis SSRI Selective serotonin reuptake inhibitor SWV Square-wave voltammetry UHPLC-HRMS Ultra-performance liquid chromatography high-resolution mass spectrometry UPLC-MS/MS Ultra-performance liquid chromatography Mass spectrometer UV Ultraviolet detection UV-Vis Ultraviolet–visible spectrophotometry VDP Differential pulse voltammetry WHO World Health Organization 1 CHAPTER 1 INTRODUCTION 1.1 General background Environmental contamination is an issue of growing concern worldwide. Due to the known and unknown possible impact on the environment, Human and wildlife health, emerging contaminants have raised alarms among the scientific community and official environmental agencies. Emerging contaminants are newly developed or categorized as contaminants or known compounds freshly identified in the environment (frequently associated with analytical developments) and cause known or suspected adverse ecological and/or human health effects. They incorporate a wide variety of substances such as personal care products, pharmaceutical products, pesticides, veterinary products, heavy metals, microplastics, industrial by-products, engineered nanomaterials, food additives, and natural and synthetic hormones. Their metabolites and transformation products are also included (X. Li et al., 2022; Puri et al., 2023). After their entrance into the environment, they present several possible biogeochemical behaviors. The most common and impacting are transformation, transfer, and accumulation along the food chain, altering some properties and possibly turning their detection into a challenging process. Additionally, these changes and wide-spread can result in distinct and unknown impacts on environmental health (X. Li et al., 2022). Anthropogenic activities are the major sources of emerging contaminants’ introduction into the environment. Wastewater treatment plants (WWTPs) are one of the main entry channels of these compounds in the soil and water environmental compartment, as they receive wastewater from several sources (e.g. households, hospitals, industries) and are unable to entirely degrade emerging contaminant’s using conventional treatments (e.g. activated sludge, filtration, and disinfection) (Mukhopadhyay et al., 2022; Puri et al., 2023; Terpenning & Öberg, 2017). Additionally, according to World Health Organization (WHO), in 2020, nearly 22% of the global population (1.7 billion people) still did not have basic sanitation services, and 45% of the household wastewater globally generated was discharged without safe treatment, increasing the potential for effluents with high concentrations of these contaminants to be released into water bodies (World Health Organization & United Nations Children’s Fund, 2020). Pesticides are chemicals used to control plagues and diseases in agriculture, while pharmaceuticals are drugs used to treat or prevent diseases in humans and animals. Even though these substances have brought significant benefits to society, their widespread use Introduction 2 has also led to unintended consequences, such as soil, water, and air pollution (Sousa et al., 2018). Both pesticides and pharmaceuticals are considered emerging contaminants of concern because their impact on the environment is complex and multifaceted, and their effects on ecosystems can be subtle and challenging to predict, leading to long-term ecological and economic consequences. Due to the continuous introduction of human-made and natural organic substances into the environment, it is crucial to understand better the chemical status of the Earth's surface water and soils. To reduce and prevent water and soil pollution, it is essential to invest in infrastructure and technology to improve wastewater treatment and reduce the discharge of pollutants into water sources. Promoting sustainable practices in agriculture and industry is also crucial, as increasing public awareness of the importance of protecting our water resources. This work address and explore the presence, potential sources, pathways, and impacts of environmental contamination by emergent pollutants, focusing on four significant substances: glyphosate and its primary degradation product (AMPA) and glufosinate, pesticides from herbicides family and fluoxetine, a pharmaceutical belong to antidepressant group. Also, examine some of the strategies and technologies used to mitigate this problem proposing and assessing adsorption as one of the technologies applied to treat these contaminants in water or directly in soils, preventing them from flowing into water bodies. 1.2 Objectives and structure 1.2.1 Main objective This work aimed, firstly, to improve knowledge within the scope of new contamination trends as well as the primary sources of contamination and their pathways to reach the environmental compartments. Glyphosate, its major degradation product AMPA, glufosinate and fluoxetine were selected to represent two different families of compounds: herbicides and pharmaceuticals. Thus, also intend to fill in the gaps in the existing knowledge on glyphosate, AMPA, glufosinate and fluoxetine environmental contamination and to propose a low-cost and sustainable remediation technique to prevent their spread and consequent contamination of soils, and water resources, namely the application of tailor-made Technosol. Secondly, it sought to search for inexpensive and sustainable material with high glyphosate, AMPA, glufosinate and fluoxetine adsorption capability that could be incorporated as a tailormade Technosol component. Chapter 1 3 Figure 1.1 presents a flow chart of the several steps through the subject’s approach with a focus on the main objectives of the work. Figure 1.1 - Flow chart of the topics approached throughout the thesis to reach the objectives. 1.2.2 Specific objectives 1. To monitor sites contaminated with pharmaceuticals and pesticides, namely fluoxetine, glyphosate, AMPA and glufosinate in Portugal and in the World; 2. To discover sustainable material to use as adsorbents. Additionally, to understand the characteristics that make them more favorable to the adsorption of the target compounds; An depressants Fluoxe ne erbicides lyphosate AMPA lufosinate Literature Review 10 could be generated by their use by human and animal as well as by industry. Excretion of unmetabolized pharmaceutical drugs by the human/animal body after consumption, disposal or flushing down the toilets of unused products, and bathing, and washing off topically applied medications are three of the primary routes that promote the arrival of these contaminants to water bodies through WWTPs or direct discharge (J. L. Santos et al., 2022; Silori et al., 2022). Figure 2.3 – Therapeutical classes of the most found pharmaceuticals in environmental matrixes (developed by the author). Moreover, they continuously move into the environment through different pathways, namely their discharge from household, hospitals, or industry to wastewater treatment systems or directly to the environment. Additionally, the use of the sludges produced in WWTPs as organic amendments and nutrients source for agricultural purposes (highly frequent in European Union, with specific application rates throughout the Member States), and the irrigation with recycled water or wastewater treatment discharge, represent a probable pathway to these compounds to end up in soil or in the aquatic environment, increasing the risk of soil and water contamination (J. L. Santos et al., 2022). Thus, conventional WWTP represents a significant source of human pharmaceuticals in the environment. Since their treatment methods were not designed to remove these chemicals, and due to the pharmaceutical’s complex structure and physicochemical properties, they are ineffective in entirely removing them (J. L. Santos et al., 2022; J. Sharma Chapter 2 11 et al., 2022; Silori et al., 2022). Nevertheless, the contamination sources and the entry points into the environmental compartments can vary greatly depending on the circumstances of each country. For example, in East African countries, improper pharmaceutical handling and disposal is the main challenge (Karungamye et al., 2022). Regarding the removal of pharmaceuticals in WWTP, according to several studies, only in occasional cases was there a reduction in the concentration of these compounds in the influent compared to the effluent. This variance was highly dependent on the applied treatment levels and systems at the facility. The concentration reduction in water may not have been exclusively due to the removal of these compounds but rather to their concentration in the solid phase resulting from the various treatments (sludge). Additionally, the output concentration was higher for some pharmaceuticals than the input concentration. Concerning psychiatric drugs, no removal in the WWTP was observed (Lopez et al., 2022; Paula Paíga et al., 2016, 2019; Papageorgiou et al., 2016). In Europe, the Water Framework Directive is the primary regulation for water protection. It applies on the quality of the inland, transitional, and coastal surface waters through the “daughter” Environmental Quality Standards Directive defined in the Directive 2000/60/EC (European Commission, 2000), and on the quality and quantity of groundwater, through the Groundwater Directive 2006/118/EC (European Commission, 2006). One of the several amending of Directive 2000/60/EC, the Directives 2008/105/EC of 16 December (European Commission, 2008) and 2013/39/EU of 12 August (European Commission, 2013) stipulated that the European Commission should establish a watch list of substances for which Unionwide monitoring data would be gathered to support future prioritization exercises in agreement with the Article 16 of the emended directive, complementing its articles 5 and 8. The continuous monitoring period for any individual substance included in the list must be at least 12 months and not exceed four years. Additionally, this list must be updated every 24 months after that, and any substance for which a risk-based assessment, as referred to in Article 16 (2) of Directive 2000/60/EC, can be concluded without additional monitoring data should be removed. Following the previously established, a watch list of substances for Unionwide monitoring in the field of water policy was created, starting with 10 substances or groups of substances (as recommended in the Directive 2013/39/EU), firstly defined in the Commission Implementing Decision (EU) 2015/495 of 20 March 2015 (Figure 2.4) (European Commission, 2015b). In its annex are included some indicative analytical method for each substance or group of substances and the maximum acceptable method detection limit. Substances included or added to the watch list must be carefully chosen from amongst those Literature Review 12 that exhibit signs of potentially significant risks to or via the aquatic environment at the European Union level but for which there is no sufficient data to make a conclusive determination. The watch list was updated in 2018 (European Commission, 2018), 2020 (European Commission, 2020b), and 2022 (European Commission, 2022a). Currently, this list includes 26 substances or groups of substances, and for most of these compounds, the SPE extraction method and LC, HPLC, or UPLC in tandem with MS/MS analysis method are suggested. Figure 2.4 - Evolution of the watch list throughout the years (developed by the author). Thus, increasing awareness of the environment's negative impacts, toxicity, fate, occurrences, and transfers of pharmaceutical products is essential to enhance our knowledge and regulate them. Hawash et al. (2023) executed a major review relative to the global occurrence of pharmaceuticals and personal care products residues in aquatic systems (drinking/tap water, surface water, groundwater, and WWTPs) in the period from 2012 to 2022. The highest concentrations of pharmaceutical products in surface waters were observed in Africa and South America, with a maximum reported concentration of 272,2 μg/L (amoxicillin - antibiotic) in Nigeria, 25.4 μg/L (aspirin - NSAID), and 23.5 μg/L (nalidixic - antibiotics), both in South Africa and 127.0 μg/L (caffeine - stimulant) and 21.0 (aspirin) in Brazil. In China, a caffeine maximum concentration of 9.8 μg/L was detected, followed by 2.8 μg/L of the antibiotic’s erythromycin and oxytetracycline. In Europe, the maximum concentrations observed were 3.5 μg/L (caffeine), 3.0 μg/L (salicylic acid - NSAIDs/analgesics), and 2.05 μg/L (trimethoprim - antibiotic). From the class of antidepressant compounds, and already in a much lower concentration range, frequently detected venlafaxine was reported Sulfamethoxazole Trimethoprim enlafaxine and O‐ desmethylvenlafaxine Azole compounds Clotrimazole Fluconazole Imazalil Ipconazole Metconazole Miconazole Penconazole Prochloraz Tebuconazole Tetraconazole Dimoxystrobin Azoxystrobin Famoxadone Di ufenican Fipronil Clindamycin O oxacin Me ormin and uanylurea Sunscreen agents utyl methoxydibenzoyl ‐methane Octocrylene enzophenone‐3 17‐Alpha‐ethinylestradiol (EE2) 17 ‐ eta‐estradiol (E2) and Estrone (E1) Diclofenac 2,6‐Ditert‐butyl‐4‐methylphenol 2‐Ethylhexyl 4‐methoxycinnamate Macrolide an bio cs Methiocarb Neonico noids Oxadiazon Tri‐allate Meta umizone Amoxicillin Cipro oxacin Sulfamethoxazole Trimethoprim enlafaxine and O‐ desmethylvenlafaxine Azole compounds Clotrimazole Fluconazole Imazalil Ipconazole Metconazole Miconazole Penconazole Prochloraz Tebuconazole Tetraconazole Dimoxystrobin Famoxadone 17‐Alpha‐ethinylestradiol (EE2) 17 ‐ eta‐estradiol (E2) and Estrone (E1) Macrolide an bio cs Methiocarb Neonico noids Meta umizone Amoxicillin Cipro oxacin Chapter 2 13 in a maximum concentration of 0.128 μg/L in Spain. Also fluoxetine was frequently detected, with maximum concentrations of 0.101 μg/L and 0.025 μg/L found in China and USA, respectively, and 0.017 μg/L in Spain (Hawash et al., 2023; Y. Xiang et al., 2021). Concerning superficial waters in Portugal (Leiria, Lis river), the occurrence of 33 pharmaceuticals and metabolites between 2013 and 2014 was evaluated in river water, influents and effluents of two WWTPs located along the river (Paula Paíga et al., 2016). A detection frequency of 100% was obtained for ibuprofen, ketoprofen (both NSAIDs/analgesics), carbamazepine, and fluoxetine (both psychiatric drugs), and the metabolite salicylic acid (NSAIDs/analgesics), revealing the maximum level of 1.3 μg/L for ibuprofen and 0.53 μg/L for acetaminophen (NSAIDs/analgesics) (Paula Paíga et al., 2016). Accordantly, Acuña et al. detected 50 of the 75 monitored pharmaceuticals in the Iberian Rivers (Ebro basin) and spotted a maximum concentration of 5.1 μg/L for Iopromide (βblockers), 1.9 μg/L for gemfibrozil (lipid regulator), 1.75 μg/L for fluvastatin (statin), 1.15 μg/L for hydrochlorothiazide (diuretics) and 1.8 μg/L for valsartan (antihypertensive drugs) (Acuña et al., 2015). Fluoxetine, although not detected in high concentrations in surface water, usually has high detection frequencies (Paula Paíga et al., 2016; P. Zhang et al., 2017). After achieving superficial waters, or once in the soil, these contaminants are persistent and mobile, allowing them to infiltrate groundwater through percolation (Mejías et al., 2021; J. Sharma et al., 2022; Silori et al., 2022). While groundwater is generally considered more resilient to contamination by emerging pollutants compared to surface water, traces of several contaminants, including pharmaceuticals, have been found in groundwater systems of several countries, namely China (L. Chen et al., 2018; Ma et al., 2022), Nigeria (Ebele et al., 2020; Olaitan et al., 2017), Brazil (Montagner et al., 2019), Bangladesh, Kenya (K’oreje et al., 2016), India (B. M. Sharma et al., 2019), Spain (Gasco Cavero et al., 2023), among many others. Removing them from matrices such as rocks, soil, sediments, and the unsaturated zone proves to be a challenging task. In 2012, Lapworth et al. published a wide-ranging review highlighting the world’s widespread contamination of groundwater resources by a large variety of emerging contaminants detected at potentially environmentally significant concentrations until 2011 (excluding agricultural pesticides and their degradation products). Regarding pharmaceuticals, the most frequently reported group of compounds in the analyzed studies presented average (maximum) values of 15.1 𝜇g/L (120 𝜇g/L) for paracetamol, 9.77 𝜇g/L (110 𝜇g/L) for caffeine and 5.31 𝜇g/L (99,2 𝜇g/L) for carbamazepine. Literature Review 14 After the publication of the European Commission’s roundwater Directive 2006/118/EC (European Commission, 2006), aiming to ‘prevent and control groundwater pollution’ by several contaminants, in 2014, an amendment to its Annex II was published, highlighted that the lack of information made it impossible to create new groundwater quality standards for any pollutants (European Commission, 2014). It also emphasized the need to increase the availability of monitoring data on substances posing a risk or potential risk to groundwater bodies by determining background levels and establishing a “watch list for pollutants of groundwater”. In 2019, a voluntary Groundwater Watch List (GWWL) was implemented, ranking as compounds of current concern nine pharmaceutical and two per-fluoroalkyl and polyfluoroalkyl substances (PFAS). Four additional PFAS were shortlisted for inclusion. The goal of the GWWL was to center attention on these priority compounds until detailed information is available to set regulatory levels. This list is active and constantly evolving. As new results are collected, others will be added to or replace listed ones possibly regulated. Following the work of Lapworth et al., and aiming to understand the evolution of groundwater contamination in Europe due to improvements of the Directive 2006/118/EC (European Commission, 2006), in 2021, Bunting et al., reviewed the presence of emerging organic compounds, this time focusing in European groundwater, through the analysis of 39 published studies between 2012 and 2020. Once again, the most frequently reported category of compounds was ‘Pharmaceuticals’. In this review, the most frequently detected compounds were the anti-epilepsy drug carbamazepine, the antibiotic sulfamethoxazole, the antiinflammatories diclofenac, and ibuprofen (included in the 11 compounds of current concern of the GWWL), and the lipid regulator bezafibrate. Recently, Silori et al., (2022) also performed a systematic literature review concerning the occurrence of pharmaceuticals (among other CEC) in groundwater reported from of the second decade of 21st century in the world. Figure 2.5 highlights the contamination concentration range for six pharmaceutical classes in eight of the countries included in the reviewed research. Chapter 2 15 Figure 2.5 - Range of PPCPs' groundwater contamination (𝜇g/L) of all the therapeutic classes - Analgesics/Antipyretics/NSAIDs (AG), antiepileptics (AP), β-blockers (β-B), antibiotics (AB), lipid regulators (LR) and stimulants (ST) – sample site. * Only the maximum concentration of the given therapeutic class (Adapted with permission from (Silori et al., 2022). Copyright 2022, Journal of Environmental Management). The highest concentration was found in India, of 1 390 𝜇g/L for diclofenac (NSAID), followed by ofloxacin (antibiotic) that presented a maximum concentration of 122.7 𝜇g/L, by naproxen (NSAID) of 98.39 𝜇g/L, and by sulfamethoxazole (antibiotic) of 32.00 𝜇g/L, both in USA. Sulfamethoxazole was, also, the pharmaceutical compound with the maximum concentration reported in China (20.06 𝜇g/L). In addition, Singapore presented a maximum concentration for caffeine (stimulants) of 16.25 𝜇g/L (not included in the figure). Monitorization studies in sediments contamination are less frequent. Xiang et al. (2021) reviewed the occurrence of pharmaceuticals in the aquatic environment in China, including sediments. The detection level in literature varies but several pharmaceuticals were found in concentrations higher than 100 μg/kg (103.0 - 384.59 μg/kg), including the antibiotics ofloxacin, enrofloxacin, oxytetracycline, tetracycline, trimethoprim, azithromycin, roxithromycin, and erythromycin. The highest detected concentration was found for azithromycin (1 179 μg/kg) and norfloxacin (1 140 μg/kg). Anti-inflammatory diclofenac (278.1 μg/kg) and Ibuprofen (227.1 μg/kg), antipyretic acetaminophen (320.7 μg/kg) and stimulant caffeine (482 μg/kg) were also detected (Y. Xiang et al., 2021). Lower values have been reported in Kenya (maximum concentrations ranged 1.8 μg/kg – 47.4 μg/kg for trimethoprim, and ciprofloxacin, respectively) (Kairigo et al., 2020), as well as in Brazil (maximum Literature Review 16 concentrations ranged 0.9 μg/kg – 15.5 μg/kg for diclofenac and naproxen, respectively) (de Oliveira Santos et al., 2022). The consumption of psychiatric drugs (in which antidepressants are included) has seen a significant rise in recent years, probably due to the financial crisis in Europe, followed by the COVID-19 pandemic, which promoted adverse psychological effects and numerous psychiatric disorders and, consequently, increased their usage (Borova et al., 2014; Pazzagli et al., 2022). Considering the previously reported high frequency of antibiotic detection and the increasing use of antidepressants, it is essential to continue to monitor the various environmental compartments for contamination by these products so that regulators and governments can understand how serious the current situation is and what measures will be most appropriate to address it. Pharmaceutical manufacturers, regulatory agencies, and wastewater treatment plants managers are working to address the issue of pharmaceutical contamination in the environment. Strategies include improving the design of drugs to make them less persistent in the environment, increasing the effectiveness of wastewater treatment systems to remove pharmaceuticals, and promoting proper disposal of medications by patients (Kotwani et al., 2021). Individuals can also play and important role in reducing environment pharmaceutical contamination by properly disposing of unused medications through designated take-back programs or participating in at-home medication disposal methods recommended by the Environmental Protection Agency. Monitorizations is essential to measure the level of contamination and lead to studies of toxicity and new contamination treatments. 2.2.1 Fluoxetine antidepressant use The consumption of antidepressants has been on the rise in recent decades, predominantly due to an increase in mental health issues such as depression and anxiety (Figure 2.6). The COVID-19 pandemic further exacerbated this trend, as it led to a heightened prevalence of psychological disorders. Consequently, there has been a significant rise in the use of antidepressants, including selective serotonin reuptake inhibitors (SSRIs). Antidepressant drugs, in 2020, accounted for 4% of pharmaceutical sales in Portugal, 2.7% in Spain, 2.2% in Austria, and 1.7% in Italy, suggesting a significant cost in some European countries (OECD.Stat, 2022). This growing consumption of antidepressants may pose a potential contamination risk to the environment. When individuals excrete these compounds Chapter 2 17 and their metabolites, they can enter water bodies due to the insufficient removal rates in water treatment plants. Figure 2.6 – Defined daily dosage per 1 000 inhabitants per day of antidepressant in 2017 and 2020 by the Organization for Economic Co-operation and Development (OECD) countries (Data source: (OECD.Stat, 2022) - developed by the author). As a result, these substances are released into the environment at varying concentrations, which can have multiple and diverse effects on both terrestrial and aquatic species. Exposure to these substances can lead to consequences such as changes in genetic transcription, reproductive deficiencies, and impaired motility. Additionally, they can cause alterations in the behavior of organisms during migration and when defending against predators. Overall, the increased use of antidepressants is not only a reflection of the growing mental health challenges faced by society but also a potential environmental concern that needs to be addressed (Diaz-Camal et al., 2022; Mole & Brooks, 2019). Fluoxetine is a 2nd generation antidepressant categorized as a SSRI, acting on the central nervous system by blocking the serotonin transporter in the neuron membrane. Serotonin, is a prominent neuromodulator present in the nervous systems of both vertebrates and invertebrates, playing a significant role in various physiological processes encompassing behavior, growth, and reproduction (Correia et al., 2023; National Center for Biotechnology 0 20 40 60 80 100 120 140 160 Iceland Portugal Australia Canada Sweden Spain elgium Finland Denmark Chile reece Czech Republic Slovenia Austria ermany Norway Israel Luxembourg France T rkiye Slovak Republic Netherlands Italy Costa Rica Estonia Lithuania ungary Korea Latvia De ned daily dosage per 1 000 inhabitants per day 2020 2017 Literature Review 18 Information, 2023b). Is currently approved by the U.S. Food and Drugs Administration (FDA) indicated for both acute and maintenance treatment of major depressive disorder in patients aged eight years and older, including obsessive-compulsive disorder, panic disorder, and bulimia (Correia et al., 2023; Sohel et al., 2023). It is usually sold as Fluoxetine Hydrochloride under the trade name Prozac© (National Center for Biotechnology Information, 2023b). This antidepressant was discovered in 1974 by chemists at the pharmaceutical company Eli Lilly and authorized for commercialization by the FDA for the first time in 1987 (Correia et al., 2023). Fluoxetine is a well-established medication, the fourth most used SSRI antidepressants in the USA, with approximately 23 million total prescriptions in 2020 and ranking 25th on the list of the 200 most prescribed drugs (no accessible data found for Europe in recent years) (ClinCalc DrugStats Database, 2023). Fluoxetine has received substantial attention because of its frequent detection in surface waters and municipal effluents (Biel-Maeso et al., 2018; M. P. Gomes et al., 2022; Mejías et al., 2021; Paula Paíga et al., 2019). Given its ubiquity in the environment and these type of compounds mode of action in low doses, concerns have emerged regarding the potential impacts of fluoxetine, on aquatic organisms and their associated behavioral patterns, highlighting the importance of the applicable regulation (Bertram et al., 2018; Correia et al., 2023; de Farias et al., 2019). Further investigation is essential to: (i) clarify the extent to which fluoxetine exposure may alter the normal functioning of serotonin-mediated processes in aquatic species, between other alterations, consequently affecting their survival and overall ecosystem health and: (ii) to propose new treatment technologies. 2.2.2 Antidepressant Fluoxetine - chemical and physical properties In addition to the amount consumed and excreted, same pharmaceutical properties could define their tendency to contaminate and accumulate in water over time. These properties include their solubility, persistence, and bioaccumulation potential. Pharmaceuticals are often designed to be soluble in water to ensure that they can be easily absorbed by the body and reach their target sites. This high solubility allows them to be transported by water and be readily absorbed by aquatic organisms. Many pharmaceuticals are categorized as environmentally persistent, since stability is often considered to be a beneficial property to their use by humans. Additionally, their persistence is also associated to their continuous flow into the environment (Patel et al., 2019). Concerning the bioaccumulation potential, described as the ability of pharmaceuticals to accumulate in the tissues of organisms over time, it occurs when organisms absorb the pharmaceuticals at a faster rate than they can eliminate them. As Chapter 2 19 a result, the concentration of the pharmaceuticals in the organism's tissues and organs increases, potentially leading to harmful effects on the organism itself or predators that consume it (de Oliveira Santos et al., 2022; Patel et al., 2019). The main fluoxetine physicochemical properties are resumed in Table 2.1. Table 2.1 - Fluoxetine physicochemical properties (Data source: DrugBank and PubChem® 2022 database (National Center for Biotechnology Information, 2023d, 2023c)) Compound Glyphosate CAS 54910-89-3 IUPAC name N-methyl-3-phenyl-3-[4- (trifluoromethyl)phenoxy]propan-1-amine Chemical structure Formula C17H18F3NO Molecular weight (g/mol) 309.33 Water solubility at 25 ◦C (mg/L-1) 1.7 (almost insoluble)a LogKow 4.05 pKa 9.8 a 14x103 mg/L as Fluoxetine Hydrochloride The stability of pharmaceuticals can be influenced by the type of molecular fractions connected to the aromatic rings, such as nitro-, chloro-, and fluoro-functional groups, which typically increase the persistence of these compounds in the environment (Patel et al., 2019). However, other factors like pH and temperature, as well as biotic factors such as microflora and fauna, present in the water body can negatively impact the stability of pharmaceutical compounds, leading to their degradation. Despite this degradation process, the continuous release of large quantities of pharmaceutical residues and their slower rate of autodegradation can cause contamination levels to increase. This build-up of contamination can have harmful effects on human health, and aquatic life (Patel et al., 2019; J. Sharma et al., 2022). Fluoxetine half-life in natural waters range from 2 to 7 days according with several studies (Kosma et al., 2020; Pan et al., 2022), and can reach 183.2 days in ultrapure water (Yin et al., 2017). Literature Review 26 Figure 2.11 – Range of pesticides concentration (surface water (SW) and groundwater (GW): 𝜇g/L; sediments (SED) and soils: μg/kg) found in several studies (Data source: USA: Glinski et al., 2018; Spain: Postigo et al., 2021; Costa Rica: (Carazo-Rojas et al., 2018) Argentina: Mac Loughlin et al., 2022; Uruguay: Rodríguez-Bolaña et al., 2023; Tanzania: Materu et al., 2021;India: (Kurakalva & Aradhi, 2022);China: Xu et al., 2020; Germany: Tauchnitz et al., 2020; Malaysia: Elfikrie et al., 2020) DF: Detection Frequency; ND: under the detection limit (developed by the author). Y. Chen et al. (2018) assessed 31 target pesticides in surface water from two important rivers in China used as drinking water source and found, with almost 100% detection frequency, 8 organochlorine pesticides, 16 organophosphate pesticides and 7 synthetic pyrethroids with maximum concentrations ranging from 0.02–0.09 μg/L, 0.01–0.3 μg/L, and from 0.02–0.1 μg/L, respectively. The maximum summed pesticide concentration was found to be 1.7 μg/L (Y. Chen et al., 2018). Additionally, in a nearby river, Xu et al., (2020) detected 29 pesticides, with a maximum concentration of 0.60 μg/L for the fungicide carbendazim and the total summed concentration of 1.04 μg/L in winter and 0.79 μg/L in summer (L. Xu et al., 2020). In Argentina, surface water and sediments from a small horticultural stream concerning the presence of 40 compounds were studied by Mac Loughlin et al., (2022), that found herbicide glyphosate and its degradation product AMPA as the most frequently detected pesticides, with maximum concentrations in water of 20.0 μg/L and 4.9 μg/L, and in sediment of 1 146.5 μg/kg and 4 032.7 μg/kg. Pesticides were found in all matrices analyzed at every site sampled in 5 out of 6 sampling campaigns (Mac Loughlin et al., 2022b). In Georgia, USA, the concentrations found in surface water were within the same order of magnitude, with a maximum value of 10.5 μg/L for the herbicide metolachlora (also the most frequently Chapter 2 27 detected). The highest summed pesticide concentration was 39.67 μg/L, in summer (Glinski et al., 2018). In Uruguay, a monitoring survey was performed to investigate the presence of 88 pesticides in surface water (drinking water sources). 25 of the 88 analyzed pesticides were detected in surface water, with higher occurrences in spring and summer than in autumn and winter. Herbicide glyphosate, AMPA, and the insecticide chlorantraniliprole were the most frequently detected and presented values ranging 0.23-1.05 μg/L, 0.58-0.89 μg/L and 0.008- 0.063 μg/L, respectively (Rodríguez-Bolaña et al., 2023). Carazo-Rojas et al. (2018) studied the Tempisque river basin, in Costa Rica, and reported the presence, with the highest concentrations detected in the water samples among other pesticides, of the insecticide dimethoate (0.4-61.2 μg/L), and the herbicides propanil (30.6 μg/L), diuron (0.1-22.8 μg/L) and terbutryn (0.05-4.8 μg/L). The fungicide carbendazim was the most frequently detected pesticide. In sediments, the most frequently detected pesticide was the insecticide endosulfan, and the highest levels were found to be of the insecticides triazophos (0.04-491 μg/kg), cypermethrin (0.8-71.5 μg/kg) and permethrin (47.8 μg/kg), followed by herbicide terbutryn (3.0-38.7 μg/kg) (Carazo-Rojas et al., 2018). The authors also refer, as Tauchnitz et al. (2020) in Germany, that the agricultural practices near the studied water body, which involve high application rates and frequent use of some pesticides, do not appear to have a direct correlation with higher detection frequencies or concentrations found. This implies that other aspects, such as mobilization/transport, maybe more significant in determining the occurrence of pesticides in the region. Also, in Tanzania, Materu et al., (2021) found maximum levels of 3.9 μg/L (herbicide monuron) in waters, 1 830 μg/kg (insecticide chlorpyrifos) in sediments and 550 μg/kg (herbicide glyphosate) in soils and in Malaysia, Elfikrie et al., (2020) has detected fungicide propiconazole as the pesticide with the higher concentration in the studied river water, with a value of 4.5 μg/L. In general, in the analyzed studies, and as expected, the levels of pesticides found in surface waters exhibit seasonal patterns, with greater concentrations detected during periods of intensive usage (such as spring and summer, during the preparations for cultivation), and lower levels during the winter and after the harvesting of crops. Some of the detected compounds are banned compounds according to the country legislation (e.g. banned in Uruguayan: p,p′-DDT, p,p′-DDD, p,p′-DDE, atrazine, alpha-BHC and ethion; not approved for use in the European Union: acetochlor, bifenthrin, chlorpyrifos, diazinon, metalaxyl, permethrin, and simazine). This embodies a key challenge as these Literature Review 28 substances have been banned for their adverse effects on the ecosystem and human health (Rodríguez-Bolaña et al., 2023). 2.3.1 The use of glyphosate and glufosinate‐based herbicides Glyphosate or N-(phosphonomethyl) glycine, is a broad-spectrum herbicide widely used throughout the world. Comparable in structure to other organophosphate pesticides, this compound differs in terms of its toxicology, as it does not inhibit cholinesterase activity (IARC, 2017). It is used to control a wide variety of annual and perennial weeds in many crops, including corn, soybeans, cotton, and wheat. Agriculture, forestry, landscaping, and managing roadside vegetation also represent some of the glyphosate applications. When absorbed by the green parts of the plant and its root system (minority), it suppresses the function of the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), which is an intermediary in the synthesis of aromatic amino acids via the shikimic acid pathway. These amino acids are vital to synthesizing proteins and producing secondary plant products including phenolic compounds, lignin, and various growth promoters and inhibitors. Besides photosynthesizing organisms, essentially plants and algae, and some microorganisms in the soil (bacteria and fungi), EPSPS is not expressed in mammals and other vertebrates, apparently keeping them safe from unintentional damage (Fogliatto et al., 2020; Padilla & Selim, 2020; Solomon, 2020). Accidently discovered by Dr. Henri Martin in 1950 as a potential pharmaceutical, glyphosate’s potential for use in agriculture was only recognized around 20 years later, when Dr. John Franz, the founder of Monsanto Company, revealed its capabilities as an herbicide (timeline presented in Figure 2.12). The patent for glyphosate was filled in 1972, and it was first introduced to the market in 1974 by Monsanto Company under the trade name Roundup®. Though, despite its efficiency as a nonselective, broad-spectrum herbicide, the global use of glyphosate has, firstly, increased slowly over time. Due to its initial limited applications, since it can only be applied to kill all vegetation in non-crop areas such as public or private parks, cemeteries, gardens, and along roads and railway tracks, and its use in agriculture was restricted to post-harvest or late-season applications, normally applied when other control measures have failed (Benbrook, 2016; Mesnage & Antoniou, 2017). In 1996, the Roundup Ready crop’s introduction, a variety of crops genetically engineered to survive glyphosate-based herbicides application, led to a significant increase in the sales and usage of glyphosate, simplifying weed management processes (Benbrook, 2016; IARC, 2017; Mesnage & Antoniou, 2017). Chapter 2 29 Figure 2.12 – Timeline displaying the highlights of glyphosate history; GE: genetically engineered; GBH: glyphosate-based herbicide; GRG: Glyphosate Renewal Group; WHO: World Health Organization (Data source: (EFSA, 2023; European Commission, 2023; Kudsk & Mathiassen, 2020) - developed by the author). Subsequent to the introduction and spread of Roundup Ready crops use, the expiration of Monsanto’s patent and the consequent development of new glyphosate formulations by other companies, led to an exponential growth in glyphosate agricultural application, and consequent increases of its residues presence in food (J. Xu et al., 2019), feed (Zhao et al., 2018), and environmental matrix as air (Ramirez-Haberkon, Aimar, et al., 2021), soil (V. Silva et al., 2018, 2019), and water (Suciu et al., 2023). Following the World Health Organization's reclassification of glyphosate as probably carcinogenic (Group 2A) to humans in 2015 by the IARC (EFSA, 2015; IARC, 2017), the amount of published articles related to several glyphosate approaches has been growing, and numerous review articles have been published on various glyphosate-related topics and its contentiousness (Gillezeau et al., 2019; Muñoz et al., 2021; Rivas-Garcia et al., 2022; Solomon, 2020; J. Xu et al., 2019). In 2017, the Appeal Committee on Plant Protection Products legislation, involving representatives from EU member states, voted in favor of the EU Commission's proposal to re-approve the use of the herbicide glyphosate for a period of five years. However, due to intense discussions during the re-approval process, the responsibility for any further restrictions on glyphosate's use was delegated by the EU to individual Member States, contrary to the common harmonization strategy (European Commission, 2017; Huhn, 2018). lyphosate was approved rst me at the EU level R applies for license renewal EFSA announced a signi cant delay in the release of its conclusions leading to an extended for another year European Commission proposes a 15‐year renewal of glyphosate lyphosate s EU market license is renewed for 5 years Discovery of glyphosate N‐(phosphonomethyl) glycine by the Swiss chemist, Dr. enri Mar n. lyphosate was patented as an herbicide by Monsanto Introduced into the pes cides market in 1974 as a pes cide ac ve principle ‐ trade name Introduc on and legaliza on of several E herbicide‐tolerant crops ‐ Monsanto s rst patent expired Price decrease and intensi ca on of the generic prepara ons sales Reclassi ed as probably carcinogenic ( roup 2A) to humans by W O/IARC 1950 1974 2015 20172023 1972 1991 1996 2022 2016 2019 2002 Literature Review 30 While several countries acknowledge the potential hazards of glyphosate, they currently lack safer alternatives and other weed management technologies that could provide an economically viable substitute for weed control (Alcántara-de la Cruz et al., 2021; Fogliatto et al., 2020). Known alternatives to glyphosate are currently limited, less effective, and more expensive. As a result, successful and profitable weed management in major agronomic crops without glyphosate represents a significant challenge and requires the acquisition of new knowledge and skills for a successful transition (Alcántara-de la Cruz et al., 2021; Beckie et al., 2020). These discrepancies have sparked widespread controversy and led to different regulations worldwide, ranging from complete bans to permissive policies (Rivas-Garcia et al., 2022). The re-approval of glyphosate produced an intense debate regarding its probably carcinogenic effects. However, this IARC classification was opposed by other national agencies and organizations associated with the WHO, as well as the EU. This discrepancy caused a great deal of controversy surrounding the re-approval process, with the safety of glyphosate being a major point of disagreement (Huhn, 2018; Rivas-Garcia et al., 2022). Aminomethylphosphonic acid (AMPA), a weak organic acid with a phosphonic acid group, is the primary degradation product of glyphosate. In addition, AMPA is also used in the production of amino-methylene-polyphosphonates, which are a type of phosphonates widely utilized in both household and industrial applications as detergents, flame retardants, anticorrosive, anti-scaling agents, and complexing agents (mainly in the textile industry), between other industrial applications. Nevertheless, the domestic contribution to urban loads of AMPA is negligible (Grandcoin et al., 2017). Glufosinate (2-amino-4-(hydroxymethylphosphinyl)butanoic acid or D,L- phosphinothricin), also used as glufosinate-ammonium, is a non-selective herbicide with a broad weed control spectrum, also, an organophosphorus herbicide, used mostly for controlling weeds and unwanted plant growth. It is absorbed through the leaves and can cause leaf chlorosis and necrosis by inhibiting the enzyme glutamine synthetase (Takano & Dayan, 2020; C. Zhou et al., 2020). Introduced in the early 1990s, and found as a natural product, glufosinate was referred to as “a natural amino acid with unexpected herbicidal properties” (Hoerlein, 1994) and its use has a significant increase in the United States (US) over the past decade associated to the growing number of glyphosate-resistant weeds, with glufosinate serving as an alternative herbicide to glyphosate. Although glufosinate-resistant crops were available while glyphosate-resistant crops, they were less applied due to glufosinate’s lower efficacy and consistency in comparison to glyphosate. Nevertheless, the use of glufosinate is Chapter 2 31 widespread in some regions such as North America, particularly in the Midwest and Southern US where glufosinate-resistant soybean and cotton are commonly grown. In South America, it is also widely used, especially in no-till cropping systems where glufosinate-resistant cotton is popular in northeast Brazil. Other crops such as rice, orchards, vineyards, and minor crops, as well as non-agricultural areas, also account for a significant portion of glufosinate use in the western United States and other parts of the world (Takano & Dayan, 2020). Concerning European countries, glufosinate is no longer applied because its market license expired in 2018 and its use was not re-approved by the European Commission due to concerns about its toxicology, being legally banned in Europe in 2020 (European Commission, 2020a, 2022a; C. Zhou et al., 2020). Additionally, its use and selling had already been prohibited in France since the end of 2017, due to its potential toxicity to human reproduction (C. Zhou et al., 2020). Although glyphosate and glufosinate have been widely discussed in terms of their impact, the lack of publicly accessible data on their global use has resulted in increased uncertainty in the global estimates reported. Only a limited number of countries, including Portugal and Spain, make annual reports publicly available containing information about national herbicide and glyphosate/glufosinate use. Moreover, there is variability in the level of detail provided, with some reports only referring to total herbicides consumed or sold, without specifying the active ingredients. It has been observed that most of the increase in global glyphosate use can be associated with the adoption of genetically engineered herbicide-tolerant crops providing to be a solid basis to project total glyphosate use with greater confidence, in the countries where they are used. Benbrook, (2016) use this methodology to preview trends in glyphosate use in the U.S. from 1997 and 2014, being one of the most cited sources of information concerning the quantities used and, consequently, the potential for contamination. Concerning Europe, in 2019 European Network for the Durable Exploitation of crop protection strategies (ENDURE) network started a survey on the uses of glyphosate and the existing alternatives in 32 countries: the EU-28 countries plus Norway, Serbia, Switzerland, and Turkey (EU 28+4) (C. Antier et al., 2020). The main conclusions were that glyphosate sales were highest in France, Poland, Germany, Italy, and Spain (with Portugal in 10th place), that, on the other hand, considering the use of glyphosate per hectare of agricultural area, Denmark, Poland, Netherlands, Portugal, and France presented the highest values in 2017, and that glyphosate is essentially consumed by the agricultural sector (90% of total national glyphosate sales in the 13 countries where this information was available) (C. Antier et al., 2020). Figure 2.13A shows the annual total values, concerning the years 2017 and 2019, for the agricultural use of glyphosate in some European countries (some of the countries with the Literature Review 32 highest expression in the use of glyphosate according to the ENDURE report (C. Antier et al., 2020)) as well as the amount of all the other herbicides used in each country and in each year. Additionally, data on the use of glyphosate and other herbicides in the same years in Brazil and the USA, as well as in a total of 28 European countries plus Norway, Switzerland, and Turkey are shown in Figure 4B (information sources shown in the figure legend). Figure 2.13 – Evolution of herbicides sales for agricultural use with a highlight on glyphosate from 2017 to 2019: A) in some EU countries (2017 data source: (C. Antier et al., 2020); 2019 data source: Spain: (Ministerio de Agricultura Alimentación y Medio Ambiente, 2021); Portugal: (DGAV, 2021); France: (Christine Veyrac, 2021); Germany: (Federal Office of Consumer Protection and Food Safety, 2021)) and B) sum of 28 EU plus Norway, Switzerland, and Turkey (2017 – data source: (C. Antier et al., 2020); 2019:(FAO, 2022)) compared with Brazil (IBAMA, 2022) and USA (IBAMA, 2022) (both 2017 and 2019) (EU 28+3 data about 2019 glyphosate use was not found) - developed by the author. As shown in Figure 2.13A, in European countries there was a decline in the use of herbicides from 2017 to 2019, except for Portugal where a slight increase occurred (17%). Spain presented the most significant decrease, showing a reduction of about 62% in herbicide sales, assisted by a 10% reduction in glyphosate sales, followed by France, with a 26% reduction (35% in glyphosate sales), and finally Germany, with 17% overall reduction and 35% in glyphosate sales. In Figure 2.13B it can be seen that the global trend in the EU was a decrease of 11% in herbicide sales (data for glyphosate sales in 2019 for the EU28+3 were not available), in contrast with Brazil, which showed an increase in global sales of 17% and 21% in the case of glyphosate sales. The U.S. showed the same amounts of herbicides used in both years, with a 3% decrease in glyphosate sales. For the use of glufosinate, access to information is even more difficult. With regard to the amount used, this active substance represented less than 0.05% of the total herbicides applied in Spain, France, and Germany. In Brazil, it represented 0.5% in 2017 and 0.3% in 2019, and for the U.S. 2.8% in 2017, the estimate for 2019 is not yet available for the latter. Both glyphosate and glufosinate have been the subject of controversy and regulatory scrutiny in recent years due to concerns about their potential impact on human health and 0 50 100 150 200 250 300 350 400 2017 2019 2017 2019 2017 2019 Eu 28 3 razil USA 0 5 10 15 20 25 30 35 2017 2019 2017 2019 2017 2019 2017 2019 Spain Portugal France ermany Thousand tonnes of glyphosate lyphosate Other herbicides Chapter 2 33 the environment. However, both herbicides are still widely used around the world, and their safety and efficacy continue to be evaluated by regulatory agencies and scientific researchers (Z. Li & Fantke, 2022; Novotny, 2022). 2.3.2 Chemical and physical properties of glyphosate and glufosinate To ascertain the environmental fate and behavior of agrochemicals, it is imperative to consider their physicochemical properties. Key parameters in this evaluation include the organic carbon-soil partition coefficient (KOC), octanol-water partition coefficient (KOW), vapor pressure, solubility, and persistence, as represented by the time required for 50% of the parent compound to undergo transformation and dissipate from soil, water, or air (DT50) (von Mérey et al., 2016). Glyphosate and AMPA share similar physicochemical properties as its low vapor pressure, making its volatilization from soil insignificant, polarity and its moderate to high solubility in water (see Table 2.2) (EFSA, 2005, 2015). Glyphosate is a colorless, odorless, crystalline solid chemical with a three-polar functional group structure namely, amine, carboxylate and phosphonate, amphoteric and characterized by a linear arranged configuration (Ojelade et al., 2022; Spengler et al., 2021). Being a zwitterion, with four characteristic pKa values, have an ionic form, a highly polarity and solubility due to the existence of this functional groups in its structure. Despite their water solubility, glyphosate and AMPA show a high affinity for soil particles, particularly in clay-rich soils. Numerous laboratory experiments have demonstrated a significant absorption capacity of the glyphosate molecule in soil, with absorption constants ranging from 8 to 377 dm3/kg (López-Vázquez et al., 2023; Ojelade et al., 2022). Glufosinate is a phosphinic acid belonging to the organophosphorus chemical family, unlike any other chemical class of herbicides. Glufosinate is a hydrophilic herbicide (logKow: −4.0), also highly water soluble, with a low vapor pressure and with three different ionization constant (pKa) values associated with the presence of the amine and hydroxyl groups in its structure (see Table 2.2). Literature Review 34 Table 2.2 – Physical-chemical properties of glyphosate, AMPA and glufosinate (Data source FAO and PubChem® 2022 database (National Center for Biotechnology Information, 2023d, 2023c)) Compound Glyphosate AMPA Glufosinate CAS 1071-83-6 77521-29-0 51276-47-2 IUPAC name N-(phosphonomethyl)glycine (Aminomethyl)phosphonic acid 2-amino-4- [hydroxy(methyl)phosphoryl] butanoic acid Chemical structure Formula C3H8NO5P C7H10N2O4 C5H12NO4P Molecular weight (g/mol) 169.07 111.04 181.13 Water solubility at 25 ◦C (mg/L) 1.0 × 106 1.0 × 106 1370 Organic solvent solubility at 25 ◦C (mg/L) Practically insoluble Practically insoluble ≈1×10-6 LogKow -3.4 -2.17 -4.0 pKa pKa1=0.8, pKa2=2.3, pKa3=5.6, pKa4=11.0 pKa1=1.8, pKa2=5.4, pKa3=10.0 pKa1=2, pKa2=2.9, pKa3=9.8 Koc 2600-4900 1.937 9.6-1229 Vapor pressure at 25 ◦C (atm) 1.29 × 10-10 (No volatility) 1.66 × 10−7 1.16 × 10−12 (No volatility) Usually is present as an ammonium salt (Figure 2.14) in the existing preparations, which, in aqueous solution, tends to dissociate. Figure 2.14 - Glufosinate (ammonium salt) molecular structure. Chapter 2 35 2.3.3 Glyphosate and Glufosinate - degradation and dissipation paths Given the current understanding of glyphosate, glufosinate and AMPA persistence, mobility, leaching, and travel time it's possible to account for their presence in groundwater through various pathways, including point source contamination, favorable soil physicochemical properties, shallow groundwater, higher watersheds dimension, macropore flow, and the inflow of surface water or bank filtrate. Also, the existence of subsurface tile or storm drains, of sewers, and the occurrence of overland flow from developed and arid landscapes have been associated with sites with higher concentrations of these compounds (Medalie et al., 2020; Suciu et al., 2023). The prolonged utilization of these herbicides over the past 45 years has raised apprehension regarding its potential impact on health and the environment. The process of manufacturing, transporting, and applying are believed to be the major weaknesses, being mostly associated to such concern (Meftaul et al., 2020). As previously mentioned, the pervasiveness of glyphosate and AMPA across various substrates such as soil, surface/groundwater, and air has been extensively recorded. Moreover, it was even detected in human urine in several countries (Campbell et al., 2022; Nova et al., 2020; Suciu et al., 2023), indicating the extent of exposure that both humans and animals are subject to. The exceedingly low vapor pressure of glyphosate and AMPA renders it resistance to significant volatilization, even in the event of processes as mineralization/degradation, immobilization, or leaching occurred following its application. In soils these two compounds are mostly degraded by microorganisms, which use these as nutrient sources. Glyphosate could be also subject, in minor extent and in specific environments, to photooxidation and abiotic degradation(Samson-Brais et al., 2022). Upon mineralization /degradation, the breakdown products include AMPA, the primary and major metabolite of glyphosate degradation, sarcosine, and glycine (Figure 2.15). The most frequent pathway occurs through the C-N bond oxidation, in which microorganisms degrade glyphosate using it as carbon source, producing AMPA and glyoxylate (an easily assimilable energy source). Subsequently, AMPA also could undergoes mineralization, producing methylamine or formaldehyde and phosphate (Castrejón-Godínez et al., 2021; Espinoza-Montero et al., 2020; Ojelade et al., 2022). Literature Review 42 frequently used technique, applied to several sample matrix types, including aqueous samples, biological and clean-up of solid samples, although SPE cartridge are expensive and the method uses organic solvent during preparation and elution (Cheng et al., 2011; Pires et al., 2020). To water sample pre-concentration, lyophilization can also be applied (Pires et al., 2020, 2023). After the use of these procedures, if it was necessary, two types of methodologies are commonly used: direct methods and those involving a derivatization step. Direct methods are advantageous due to their reduced sample preparation steps. However, the highly polarity of glyphosate, glufosinate, and AMPA make it impossible to directly separate these compounds through reverse-phase chromatography. Some analytical columns can provide better retention of analytes with this strongly anionic phosphonate structure. Recently, several chromatographic columns have been developed to measure these pesticides, seeking greater sensitivity, reproducibility and reducing the work associated with sample preparation. Among the indirect methods used, three types of derivatization steps are most commonly applied. The first is pre-column derivatization using FMOC-Cl in ACN reagent to diminish their polar character, facilitating chromatographic retention, and allowing analysis by liquid chromatography with mass spectrometry (Figure 2.18). Figure 2.18 – Example of a pre-column derivatization of glyphosate, AMPA and glufosinate with FMOC (adapted with permission from (Campanale et al., 2022). Copyright 2022, Journal of Chromatography A). The second is post-column derivatization, promoting the conversion of glyphosate into glycine and subsequent derivatization with o-phthalaldehyde, followed by a fluorescent detector. Derivatization with trifluoroacetic acid anhydride and heptafluorobutanol, is also OO PN O O O O PN 2 O O p 9 Cl O OCl O O O O P N O O O OO P O O N 2 O O O O P O O N O O O PN O O Chapter 2 43 applied to obtain volatile and stable molecules required for gas chromatography analysis (IARC, 2017; Marta Małysiak and Tomasz Kiljanek, 2022). 2.4 Innovative technologies for mitigation of new contaminants Given the high frequency and persistence of the compounds under study in the environment and their potential impact on human and animal health, it has become essential to develop treatment processes to remediate environmental polluted matrixes. Soil and water remediation technologies are the methods used to clean up contaminated areas contained in these two environment compartments. These technologies can differ for distinct contaminants and types of contaminated matrix (soil or water). Various techniques including physical, chemical, thermal, and biological methods could be utilized for the remediation of pharmaceutical and pesticide contaminations in water and/or soil (Narimani & Silva, 2020). Physical methods mainly focus on transferring the compounds from the aqueous phase to the solid phase. On the other hand, biological and chemical treatments involve chemical reactions with the compounds, leading to the formation of new metabolites or degradation products, with complete mineralization being a potential outcome (Narimani & Silva, 2020; Patel et al., 2019; Samal et al., 2022). Some of those technologies applied to remediate pharmaceuticals and pesticides contaminated soil, water, or both are presented in Figure 2.19. Literature Review 44 Figure 2.19 – Resume of some remediation technologies applied to pharmaceuticals and pesticides contaminated soils, water, or both (developed by the author). Adsorption, electrodialysis, evaporation, dialysis, filtration, flocculation, reverse osmosis, sedimentation, and stream stripping are some of the available physical treatments, being adsorption and membrane separation the most used technologies for pharmaceutical and pesticides removal (Loganathan et al., 2023; Patel et al., 2019). Chemical methods include ionexchange, neutralization, reduction, precipitation, calcination, and advanced oxidation processes (AOPs), this last one broadly used in tertiary treatments in WWTP (Loganathan et al., 2023; Patel et al., 2019). Thermal methods essentially include incineration and pyrolysis. Concerning biological methods, activated sludge, aerated lagoons, anaerobic digestion, trickling filters, and waste stabilization ponds are used (Patel et al., 2019). The activated sludge process, based on microbial degradation, is the primary applied biological process, largely used as a secondary treatment process in WWTPs (Loganathan et al., 2023). Many of the traditional methods have been employed to remove organic substances and suspended solids from wastewater. However, some proved to be inadequate for eliminating pharmaceuticals, particularly if they are hydrophilic, and pesticides. The need for an additional tertiary treatment process arises to effectively reduce the concentration of pharmaceuticals and Chapter 2 45 pesticides in order to mitigate their potential entry and accumulation in surface water and groundwater resources (Pradhan et al., 2023). Membranes are widely used as permeable or semi-permeable barrier that selectively allows certain substances to pass through while restricting others based on their properties such as size and charge and in a driving force. The driving forces required to promote the movement of substances across the membrane can include pressure difference, concentration gradient, and a potential field that initiates ions movement (Othman et al., 2022; Patel et al., 2019). Reverse osmosis, microfiltration, nanofiltration, and ultrafiltration are effective methods using membranes for removing antidepressant pharmaceuticals and synthetic organic contaminants such as organophosphate herbicides. Concerning reverse osmosis, the driving force is the pressure difference between the filtrate and feed sites of the separation membrane. On the other hand, the primary operational driving force of nano- and ultrafiltration is ion repulsion, since the membranes used are charged. Dalbosco et al. (2023) tested reverse osmosis and nanofiltration in fluoxetine removal from water. Nanofiltration presented a removal rate between 50 and 60%, and reverse osmosis demonstrated a rate of 98.8% (Dalbosco et al., 2023). The same two technologies were also tested in Non-steroidal anti-inflammatory drugs (NSAIDs) from groundwater demonstrating successful removal rates of over 85% (Patel et al., 2019). Although its efficiency, reverse osmosis can be expensive and requires regular maintenance. This physical treatment presents the advantage of does not lead to the development of contaminant byproducts (metabolites), as technologies such as AOPs or biological-based treatments, where these transformation products could also be of environmental concern (Rodriguez-Mozaz et al., 2015). The employment of phytoremediation (i.e., the use of plants to absorb and detoxify chemical contaminants), phycoremediation (i.e., the use of algae to remove contaminants such as heavy metals, organic compounds, and nutrients, from contaminated water or soil), and microbial remediation technologies (i.e., the use of microorganisms such as bacteria, fungi, and cyanobacteria) are achieving prominence as a more cost-effective and environmentally sustainable alternative to traditional physical and chemical methods for removing heavy metals and organic contaminants, although it is a slower methodology (Bhat et al., 2020). Phycoremediation was tested using living and non-living microalgae to the removal of fluoxetine from aqueous solutions obtaining 100% of removal in six days (Andreia Silva et al., 2022). Adsorption techniques have been a key methodology in water and wastewater treatment for a prolonged period, and their growing popularity corresponds, besides to their simple Literature Review 46 design, low cost, flexibility, easy to use, and insensitivity to toxic contaminants, with the ongoing emergence of novel micropollutants in aquatic environments as well as the discovery of sustainable and cost-effective adsorbents (Afzal Husain Khan et al., 2022; Morone et al., 2019; Samal et al., 2022). Several AOPs have been implemented to eliminate pesticides, including glyphosate, namely Fenton and Photo-Fenton processes, ozonation (O3), photocatalysis, photolysis with hydrogen peroxide and UV light (H2O2-UV), electrochemical oxidation, and photoelectrochemical processes (Espinoza-Montero et al., 2020; Feng et al., 2020; Kudzin et al., 2019). Ozonation is a promising AOP that applies ozone for the treatment of wastewater and soil. Ozone is a highly potent oxidant, possessing one of the highest oxidation potentials of any known substance. This versatile agent can be applied in either a gas or aqueous phase to facilitate a wide range of chemical reactions and is capable of oxidizing pesticides, and other contaminants. Furthermore, the in-situ generation of ozone is a notable advantage, as it can rapidly decay and be converted into oxygen via chemical decomposition, thereby minimizing the buildup of toxic byproducts and promoting aerobic biodegradation. (Martínez et al., 2022). Treatment of concentrates contaminated with glyphosate and AMPA (obtained from a membrane system in a drinking water production plant) was tested applying three methodologies: ozonation, adsorption, and the combination of both. Complete degradation of both glyphosate and AMPA was observed for the highest applied ozone concentrations, mostly by direct ozonation. Adsorption onto activated carbon, on the other hand, was significantly low. These results were associated with glyphosate and AMPA high solubilities, the pH of the concentrate (8), and high natural organic matter concentrations. Adsorption followed by ozonation allowed the elimination of glyphosate and AMPA that cannot be removed by carbon adsorption. Additionally, other important advantages of this combination were lower operational costs due to reduced contact time (10 min versus 25 min) and equipment needs (Bozkaya-Schrotter et al., 2008). (Rezaei Kalantary et al., 2022) tested the removal capacity of a conventional drinking water treatment plant (WTP) with primary sedimentation, coagulation, flocculation, clarifier, rapid filtration (with sand and granular activated carbon), followed by chlorine disinfection. This study concluded that most of the pesticide’s removal occurred in coagulation-flocculation and rapid sand filtration units (87%), outcomes associated with the hydrophobic nature, and in a filtration unit using granulated activated carbon. A good performance of the total pesticides removal from the water was achieved (50%), but the removed pesticides during the coagulation-flocculation process were accumulated into the sludge that, possibly, will be Chapter 2 47 discharged into the environment without any treatment, ending up being released into the soil (Rezaei Kalantary et al., 2022). In the environment, the main dissipation process of glyphosate from the soil is through biological breakdown, primarily driven by microbial activity, including actinomycetes, bacteria, fungi, and microcytes, in which bacteria play a dominant role. The microbial degrading capacity has been extensively studied and reviewed, and it is well established that bacteria can use it as a source of inorganic phosphorus, as well as nitrogen and carbon sources under certain conditions (Singh et al., 2020; Zabaloy et al., 2022; Zhan et al., 2018). Therefore, it is imperative to consider soil habitats as a crucial factor that regulates not only the biodegradation of glyphosate but also its adsorption and dissipation, thereby contributing to its overall presence in the environment (Zabaloy et al., 2022; Zhan et al., 2018). Soil vapor extraction is a soil remediation technique that removes volatile contaminants, using a vacuum pump, but it can be costly and time-consuming (Baldissarelli et al., 2019). On the other hand, enhanced bioremediation is a treatment that implies adding nutrients to the soil to increase specific microorganisms’ growth capable to degrade contaminants. This is a cost-effective and environmentally friendly technology, but time-consuming until the desired results are achieved (Pathak et al., 2022). Concerning electrokinetic remediation, in which an electric field is applied to the soil to promote the contaminants removal, is more effective for removing heavy metals and organic contaminants (as some pesticides), but it can be expensive and energy-intensive (Baldissarelli et al., 2019; F. L. Souza et al., 2016). For chemical oxidation treatments, the addition of an oxidizing agent is performed (to contaminated soil or water) resulting in the contaminant’s oxidation. Effective for treating a wide range of contaminants, but it can be expensive and requires careful monitoring to avoid secondary contamination. Adsorption can also be applied to soil and water and is a process by which a solid material captures and holds molecules of a dissolved or suspended substance from a liquid or gas (Samal et al., 2022). This technology is widely used to remediate contaminated sites, as it can effectively remove a variety of pollutants. The quest for novel methods to eliminate different pollutants in wastewater is an ongoing endeavor. However, it is equally important to establish that the treated matrices are environmentally safe, as any compound can pose a threat as a pollutant, depending on its concentration and the site where it occurs. Therefore, nonstop research and development of novel methods hold significant importance for the future elimination of a range of emerging contaminants (Diaz-Camal et al., 2022). Literature Review 48 2.4.1 Adsorption Adsorption is a process by which a solid material, called an adsorbent, captures, and holds onto molecules of a dissolved or suspended substance, called an adsorbate, from a liquid or gas. This is an interface process that involves physical interactions, such as electrostatic forces, and/or chemical bonds to attach species (adsorbates) from the gas or liquid phase to a solid material (adsorbent) (Sidoli et al., 2016). This process is widely used as a means to remediate contaminated sites or treat contaminated effluents, as it can effectively remove a variety of pollutants from water or soil (Espinoza-Montero et al., 2020; Morone et al., 2019). Adsorption has several advantages for the remediation of the contaminated matrix. It can effectively remove a variety of pollutants, including heavy metals, organic compounds, as well as nutrients. It can, also, be used to treat groundwater, surface water, effluents, and soils. It is relatively simple to operate, with high removal rates of most pollutants and a cost-effective method compared to other remediation techniques (Espinoza-Montero et al., 2020; Sidoli et al., 2016). Contaminants that can be removed by adsorption include pesticides, herbicides, and industrial chemicals (J. Chen et al., 2017; McGinley et al., 2022; Mojiri et al., 2020). Adsorption is also effective in removing lead, mercury, and arsenic (Guo et al., 2020). The effectiveness of adsorption depends on several factors, such as the type of adsorbent used, the concentration and type of contaminant, and the contact time between the adsorbent and the contaminated liquid or gas (Goldan et al., 2022). In the case of water treatment, adsorption can be used as an individual technique or as part of a multi-step treatment process, such as in combination with biological or chemical treatment methods (Mukhopadhyay et al., 2022). The most used adsorbents for the remediation of contaminated sites are activated carbon, biochar, zeolites, and clays. These materials have high surface areas and pore volumes, which allows them to adsorb many pollutants from the surrounding environment (Amalina et al., 2023; McGinley et al., 2022; Rissouli et al., 2017; asiljević et al., 2019). In addition, they can be easily modified to improve their adsorption capacity for specific pollutants (Tee et al., 2022). The effectiveness of adsorption for remediation of contaminated sites depends on several factors, including the type and concentration of pollutants, the properties of the adsorbent, and the site conditions. In addition, the adsorption capacity of the adsorbent can be affected Chapter 2 49 by factors such as temperature, pH, and the presence of competing ions (G. Oliveira et al., 2018). Adsorption can be used in several ways for remediation of contaminated sites, including in-situ and ex-situ treatment. In-situ treatment involves treating the contaminated soil or water on-site, while ex-situ treatment involves removing the contaminated soil or water from the site for treatment. In-situ treatment is generally preferred, as it is more cost-effective and less disruptive to the surrounding environment. It could also be applied as a WWTP treatment, being included in the developed treatment process (Espinoza-Montero et al., 2020; Mukhopadhyay et al., 2022). The extent of adsorption significantly influences the transfer and availability of contaminants as pesticides in soil and is one of the main mechanisms that control pesticide leaching. This process reduces the ability of pesticides to migrate and limits their potential to contaminate surface waters or groundwater, making it a crucial factor in preventing environmental pollution (Sidoli et al., 2016). In summary, adsorption is a promising technique for remediation of contaminated sites. It is a simple and cost-effective method that can remove a wide range of pollutants from water and soil. However, careful consideration of site conditions and the selection of appropriate adsorbents are necessary to achieve successful results. 2.4.2 Prospective use of biochar to remediate contaminated sites Biochar is a type of carbonaceous material produced thru the pyrolysis of organic materials, such as agricultural waste or forestry residues in an oxygen-limited environment. The thermal breakdown of lignin, cellulose, hemicellulose, fat, and starch contained in the feedstock occurs during pyrolysis and produces three distinct results: biochar (solid fraction), partly condensed volatile matter known as bio-oil (liquid fraction), and non-condensable gases (gaseous fraction) as CO2, CO, CH4, and H2. These gases and bio-oil can be captured to generate energy and create worthwhile byproducts, such as food flavoring, biochemical compounds, adhesive, and wood preservatives, based on the feeds used (Goldan et al., 2022; Pokharel et al., 2020). It has gained increasing attention as a potential tool for remediating contaminated soil or water due to its ability to adsorb and immobilize contaminants (Lehmann & Stephen, 2009; Z. Li & Fantke, 2022; McGinley et al., 2022). Literature Review 50 The use of biochar for remediation technology, as adsorption, has several advantages. Firstly, biochar can be produced from a wide range of waste organic materials that are readily available, and acting as a recycling process, which makes it a cost-effective and sustainable solution (Rubio, 2018). Second, biochar has a high surface area, allowing it to adsorb several substances as heavy metals, organic compounds, and nutrients. Third, biochar is stable and can remain in the soil for hundreds of years, which makes it a long-term solution for contaminated sites. The last is that it is an organic carbon-rich material capable to improve soil conditions as: increases soil organic carbon content, porosity, water, and nutrient retention capacity, and develops microbial population and activity, inclusively stimulating the growth of beneficial microorganisms that combat plant diseases and acquire resistance to pests and diseases (K. Weber & Quicker, 2018). Therefore, biochar can be applied to contaminated soils through direct mixing with soil, incorporation into soil amendments, or application as a top layer. When biochar is mixed with soil, it can improve soil fertility, and water-holding capacity, promote carbon sequestration, and microbial activity, potentially enhancing plant growth (Ahmad et al., 2014; McGinley et al., 2022). Several studies have shown that biochar can be effective in remediating contaminated soils and water and applicated in distinct approaches as directly in the contaminated site or used as adsorbents in water treatment facilities (Aghababaei et al., 2023; Bilias et al., 2021; Guo et al., 2020; McGinley et al., 2022; L. Xiang et al., 2022). According to existing literature, the addition of biochar to contaminated soils can effectively immobilize heavy metals and several persistent organic pollutants, reducing their bioavailability. Precipitation, electrostatic interaction, surface adsorption, and structural sequestration, are some of the immobilization mechanisms referred to as most predominant, leading to a facilitated decomposition. However, the extent of decontamination efficacy is dependent on several factors such as the biochar source, amendment rate, soil type, and pollutant species (Guo et al., 2020). Xiang et al. (2022) reviewed the use of biochar, bacteria, and plants in the sustainable remediation of soils contaminated with organic pollutants. As shown in Figure 2.20 illustrate the potential mechanisms involved in this integrated approach, including plant uptake, bacterial degradation, and biochar adsorption, the integration of these three components can significantly enhance the removal efficiency of organic pollutants from soil. The use of such a comprehensive strategy provides a promising solution to the complex challenge of organic pollutant contamination in soil (L. Xiang et al., 2022). Chapter 2 51 Figure 2.20 - Proposed mechanisms by which biochar mediates the remediation of organic pollutants in soil. (adapted with permission from: (L. Xiang et al., 2022). Copyright 2022, Environmental Science and Technology). As highlighted in Figure 2.20, biochar facilitates organic pollutant oxidation/degradation via the production of reactive oxygen species, such as •O , SO4•-, and O2•-, through free radical reactions with O2, S2O82-, or H2O2 (Figure 2.20a). Organic pollutants can also be immobilized in soil via different biochar interactions, a process in which carbonized fraction of biochar intermediates adsorption (electrostatic attraction and nonpolar biochar–pollutant interactions), and uncarbonized faction intermediates partition (Figure 2.20b). Furthermore, biochar's porous surface can host soil microbial communities, restrain and release enzymes, and relocate electrons to microorganisms and pollutants, influencing organic pollutants metabolism of plant and microbial communities in soil (Figure 2.20c) (L. Xiang et al., 2022). However, the use of biochar for the remediation of contaminated sites has some limitations. First, the effectiveness of biochar depends on the type and concentration of contaminants in the contaminated matrix. Second, the optimal application rate and method of biochar application depend on the specific site conditions (when the application is in-site). Finally, the long-term effects of biochar application on soil properties and plant growth are not well understood and require further research (Bilias et al., 2021). Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 58 Chapter 3 59 Abstract To our knowledge, this is the first study in Portugal for the analysis of both antibiotics and psychiatric drugs in river waters from Douro and Leça rivers and its sediments. Samples were extracted using solid phase extraction and QuEChERS procedures and the analysis was performed using liquid chromatography with tandem mass spectrometry. Higher number of pharmaceuticals and higher concentrations were found in the Leça river for the two matrices under study. The highest detection frequency for the twenty-seven pharmaceuticals was observed for fluoxetine with 83.3% in both matrices. None of the studied antibiotics were detected in water collected in the Douro river however six antibiotics (azithromycin, ciprofloxacin, clarithromycin, moxifloxacin, ofloxacin, and trimethoprim) were found in the Leça river. Further, the analysis of sediments exhibits the occurrence of sulfamethoxypyridazine in the Douro river and azithromycin in the Leça river. The highest concentration was observed in Leça river for azithromycin in the river water (2,819 ng/L) and sediments (43.2 ng/g). Carbamazepine, citalopram, fluoxetine, sertraline, trazodone, and venlafaxine were the psychiatric drugs detected in river waters and sediments with more diazepam as water river contamination. Concentrations ranged from <method detection limit (MDL)) to 2.0 ng/L (river waters) and <MDL to 0.251 ng/g (sediments) in Douro river and between <MDL to 354 ng/L (river waters) and <MDL to 6.35 ng/g (sediments) in Leça river. To identify the possible sources of pollution more monitoring studies should be performed along the studied rivers. Keywords: Pharmaceuticals, QuEChERS, Monitoring, Surface waters, Sediments, Solid Phase Extraction 3.1 Introduction The presence of pharmaceuticals and its metabolites and degradation products in the environment requires continuous research and monitoring studies to assess their potential risks to the human and ecosystem’s health (Paula Paíga et al., 2019). These studies need to be performed for the assessment of pharmaceuticals in the different environmental compartments to evaluate their distribution, to identify contamination sources, and to contribute for establishing limit values to protect the environment and human health. The trend in sample preparation in trace analysis emphasized the optimization of extraction procedures for being simpler, faster, less expensive, and more environmentally friendly than Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 60 traditional techniques. The analytical methodologies are better established and nowadays, the scientific community focuses more on monitoring studies. The pharmaceuticals prevalence in the environment is currently a well-established and well-quantified problem and has become a matter of both scientific and public concern (Lolić et al., 2015; Paula Paíga et al., 2019) due to its environmental risk. Moreover, several studies have examined the bioaccumulation of pharmaceuticals and their toxicity on non-target organisms as fish, algae, crustaceans, mussels, snails, between other animals, presenting bioaccumulation factors from 1.4 to 32000 (Ebele et al., 2017). There are several possible sources and routes for pharmaceuticals to reach the environment, but wastewater treatment plants (WWTP) have been identified as the main trail (Jelić et al., 2012) Sources of wastewater include domestic, but also hospital wastewaters, medicine manufactures, landfill leachates and direct disposal of unused drugs in the environment (Nikolaou et al., 2007; Ortiz de García et al., 2013). After the treatment performed in WWTP, the treated effluents are discharged into rivers, and the produced sludges are eliminated or reused in agricultural as a fertilizer product. As a consequence of the incomplete elimination of the pharmaceuticals and metabolites in the WWTP, these compounds reach all environmental compartments (Nikolaou et al., 2007). The incidence of pharmaceuticals in the aquatic environment is con rmed at wastewater, surface water, groundwater, and drinking water (Ortiz de García et al., 2013). However, since these compounds have the ability to partition to environmental solid phases (Minten et al., 2011), increasing attention has been paid, in recent years, for the contamination of pharmaceuticals in soils all over the world. Nonetheless, sediments are natural repositories of many chemical substances present in the water (Beretta et al., 2014) and sediments provides a wide variety of binding sites and act as major sinks for the deposition of pollutants (Ortega-Calvo et al., 2013; Sakari et al., 2014). In recent years, antibiotic and psychiatrics pharmaceuticals consumption has attracted attention worldwide. As a result of the strategic plans for the reduction of antibiotic consumption in the European Union including Portugal (i.e. Program for Prevention and Control of Infections and Resistance to Antimicrobials), overall consumption of antibacterials has been declining in recent years, remaining at a still high level (21.6 daily doses defined per thousand inhabitants per day in 2016) for Portugal with similar value observed in Europe (21.9 daily doses defined per thousand inhabitants per day) (Rodrigues et al., 2018). Concerning to the psychiatric drugs and despite the implementation of the National Program for Mental Health that focus on the rationalization of its prescription and on the prevention of mental Chapter 3 61 health problems, there has been an intensification of its consumption, with an increase of 15.2 M to 30.0 M consumed packages from 2013 to 2016 (Programa Nacional para a Saúde Mental, 2017). Human survival depends on sustainable linkages with the environment. However, the "present" shows a different reality. The degradation of the environment is constant by the strong activities of the human being. Thus, stopping pollution is the most problematic subject of the planet. In the last decades, a new topic has attracted attention, the issue of pharmaceuticals and their potential environmental risk. Over the last years, the assessment of pharmaceuticals in surface waters has increased but fewer studies are being performed in soils and sediments. Thus, in the present study, the assessment of pharmaceuticals belonging to antibiotics and psychiatric drugs in surface water and sediments of two rivers (Douro and Leça) located in the North of Portugal was performed. Published studies performed by the scientific community in the Douro river showed that its pollution by pharmaceuticals is consistent and its occurrence is frequent, covering a wide area and displaying hot-spots (Lolić et al., 2015; Madureira et al., 2010). In the Leça river, data regarding the occurrence of pharmaceuticals in its basin is scarce, however, the existential pollution in the Leça river must not be neglect. Thus, solid phase extraction (SPE) for river waters collected along the Douro and Leça rivers and quick, easy, cheap, effective, rugged, and safe methodology (QuEChERS) used for in the sediments from the same rivers bank were the procedures used for the extraction of twenty-seven pharmaceuticals belonging to antibiotics and psychiatric drugs. The analysis was performed using liquid chromatography with tandem mass spectrometry (U PLC−MS/MS) and the obtained results in river waters and sediments (top and bottom) were analyzed. The environment risk coefficient was performed as also the correlation between the parameters and the obtained concentration of the studied pharmaceuticals. To our knowledge, this is the first work in Portugal that simultaneously analyses river waters and sediments for the set of pharmaceuticals under study. 3.2 Materials and methods 3.2.1 Physical characteristics and points of pollution of the Douro and Leça rivers Draining to the Atlantic Ocean, Douro and Leça rivers are two of the rivers that cross the city of Porto in Portugal. Douro river, with its source in Urbion (Spain), covers an area of about 98 000 km2 shared among Portugal and Spain, with a total length of 927 km and an average Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 62 flow of 700 m3/s. In the other hand, Leça river that rises in Monte Cordova, Santo Tirso (Portugal), covers an area of only 190 km2, with a total length of 45 km and an average flow of 3.4 m3/s. Its main information’s are present in Table SM2 (Supplementary Material). The Douro River receives effluents from several WWTPs. Two of them, located in the lower and medium stretch of the estuary, belonging to the metropolitan area of the Porto city are Sobreiras and Freixo WWTP, which together serve an equivalent population of about 370 000 inhabitants, achieving an average daily flow of about 90 000 m3 (Cruzeiro et al., 2017; Madureira et al., 2010; Mendes et al., 2017). The two most important WWTP that discharge effluents to Leça river are located in Maia (Parada and Ponte de Moreira WWTP). Those WWTP treat daily nearly 23,000 m3 per day of urban wastewaters, from domestic and industrial effluents (A. I. Gomes et al., 2014). The Leça river receives untreated and treated effluents from numerous industries, of quite different typologies, namely: textile dyeing and printing, metallurgical, mechanical, and agro-food plants (A. I. Gomes et al., 2014). 3.2.2 River waters and sediments sampling Samples (water river and sediments) were collected in November of 2017. The sampling points were numbered from Crestuma-Lever dam to the river mouth in Douro and from rural areas to the port of Leixões in Leça (Figure 3.1). Sampling sites 4 and 5 from the Douro River were located near the river mouth (approximately 1.5 km in the right riverbank), shortly after the discharge point of Sobreiras WWTP. In the same riverside, points 2 and 3, about 2 km upstream, were set in Massarelos. The first sampling point, about 4.5 km upstream, in the other riverbank (Vila Nova de Gaia), was located in the Areínho river beach. Concerning to Leça River sampling points, one was located in a recreational park, downstream the discharge of Ponte de Moreira WWTP (point 6), and point 7, in Ponte do Carro recreation area, also inserted in a revitalized zone, after a strongly industrialized area. The course of the Leça river is carried out between private agricultural fields and located in areas with reduced accessibility, limiting the access to the river. Thus, fewer sampling points were made in the Leça river. Chapter 3 63 Figure 3.1 - Sampling locations of surface water and sediment samples in the Douro and Leça rivers. River waters were collected five centimeters deep, near the margin of each river (approximately 50 cm), using a mat high-density polyethylene bottle pre-washed with ultrapure water. Sediments were collected in the same sampling points, at two depths (top: 0-2 cm and bottom: 2 to 10 cm), except at the collection point in the Ponte do Carro recreation area (point 7, Figure 3.1), since these sediments were not accessible due to the riverbed structure. A total of twelve samples (top and bottom) were collected. Subsequently, samples were kept at 4°C until arrival to the laboratory. Water samples were vacuum filtered through 0.45 μm nylon membrane filters (Fioroni Filters, Ingré, France). All the samples were stored at -20°C until extraction (no more than one week). 3.2.3 River waters and sediments characterization Water temperature (T), pH, conductivity (CE), redox potential (Eh), and dissolved oxygen (DO) were measured in situ, along the sampling campaign. Water quality parameters such as element analysis (F-, Cl-, Ca, Mg, Na, K, Fe, Al, Si), heavy metals (As, B, Cd, Co, Cr, Cu, Mn, Ni, Pb, Zn), and sulphate (SO42−), ammonium (NH4+), nitrate (NO3-), and nitrite (NO2-) ions were determined according to the procedure described in Ferreiro et al. (2017). Parameters including pH (H2O, KCl, and CaCl2) (1:2.5 m:V), conductivity (CE), water content (WC), organic matter (OM), phosphate (PO43−), and sulphate (SO42−) (Mussa et al., 2009) were analyzed in sediment, and the same element analyzed in river waters and heavy Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 64 metals analysis was performed (Ferreiro, 2017). Total Phosphorus (P) and extractable P (Olsen method) were measured, in addition to total carbon, nitrogen, and sulphur determination (analyzed by combustion with a LECO analyzer). A particle size distribution characterization was performed using a laser diffractometer (Mastersizer 2000, Malvern, UK) coupled with a dispersion unit - Hydro G, (Malvern, UK), capable to analyze particle size between 0.02 to 2000 µm. The refractive index for the dispersed phase was 1.65 and the absorbance was 0.1. The used continuous phase was water, with a refractive index of 1.330. The sample amount placed within the measurement system, with a velocity of the stirrer of 600 r.p.m. and pump speed set at 1250 r.p.m., was such that the value of obscure fell within the range of 10-20%. 3.2.4 Reagents and chemicals For the extraction, acetonitrile (assay 99.99%) and methanol (assay 100%) were purchased from VWR Chemicals (Fontenay-sous-Bois, France), hydrochloric acid (HCl) 37% were supplied by Carlo Erba (Rodano, Italy), ammonium hydroxide solution (NH3aq, 28.0– 30.0%) was obtained from Sigma-Aldrich (Steinheim, Germany), and ethylenediaminetetraacetic acid disodium salt 2-hydrate (Na2EDTA) (assay >99.0%) was obtained from Panreac (Barcelona, Spain). Acetic acid glacial (assay 100%) was purchased from VWR Chemicals (Fontenay-sous-Bois, France). For the analysis in UHPLC-MS/MS, methanol and acetonitrile, both LC-MS grade, were supplied by Scharlau (Barcelona, Spain), propanol LC-MS grade was obtained from Sigma- Aldrich (Steinheim, Germany), and formic acid (PA-ACS) was supplied by Carlo Erba (Rodano, Italy). A Simplicity 185 system (Millipore, Molsheim, France) was used for producing ultra-pure water (resistivity of 18.2 MΩ.cm). All chromatographic solvents were filtered through a 0.22 µm nylon membrane filter (Supelco, Bellefonte, PA, USA) using a vacuum pump (Dinko D-95, Barcelona, Spain) and degassed for 15 min in an ultrasonic bath (Sonorex Digital 10P, Bandelin DK 255P, Germany). All samples were filtered through a PTFE syringe filter (0.20 μm) prior to the injection in the UHPLCMS/MS system. All antibiotics and psychiatric drugs and isotopically-labelled internal standards were of high purity grade (≥98%). Lomefloxacin, moxifloxacin, fluoxetine, norfluoxetine, paroxetine, sertraline, trazodone, venlafaxine, ciprofloxacin-d8, and fluoxetine-d5 were obtained in the form of the hydrochloride salt. The chemical abstract service (CAS), molecular weight, Chapter 3 65 formula, structure, supplier company, and the solution used for prepared each stock solution are listed in Table SM1 (Supplementary Material). Individual stock standard at a concentration of 1 g/L was prepared on a weight basis in solvent (See Table SM1, Supplementary material) and stored at -20ºC and an intermediate solution (mixture of all compounds), with a fixed concentration of 20 mg/L, was prepared by adding an appropriate amount of each stock solution. Working standard solutions were prepared from the intermediate solution in acetonitrile:ultra-pure water (30:70, v/v) before each analytical run and for the fortification tests. 3.2.5 SPE cartridges and QuEChERS tube SPE cartridges Strata-X (200 mg, 3 mL) from Phenomenex (California, USA) and original QuEChERS extract tubes (1 g NaCl and 4 g MgSO4) obtained from Agilent Technologies (Lake Forest, CA, USA) were used for the extraction of waters and sediments, respectively. After QuEChERS extraction, a dispersive SPE (150 mg C18 and 900 mg MgSO4) was used for cleanup step (Agilent Technologies (Lake Forest, CA, USA)). 3.2.6 River waters and sediments extraction procedures SPE and QuEChERS extraction methodologies used were adapted from the work of P. Paíga et al. (2017) for river waters and Santos et al. (2016) for sediments, respectively. The steps used in the two extraction methods were illustrated in Figure SM1 (Supplementary material). At the end of each extraction, the extracts were evaporated under a gentle stream of nitrogen to dryness and the residues were reconstituted with 500 μL of acetonitrile:ultrapure water (3:7, v/v). Finally, 5 μL of a mixture of isotopically-labelled internal standards solutions was added. 3.2.7 UHPLC-MS/MS analysis The chromatographic and mass spectrometry (MS) conditions used for the antibiotics and psychiatric drugs under study are described in the article published by the authors (P. Paíga et al., 2017). A representative chromatogram of a 500 µg/L standard mixture of the selected pharmaceuticals is present in Figure SM2 (Supplementary material). Retention time and ion ratio for all compounds are listed in Table SM4 (Supplementary Material). The concentration of isotopically-labelled internal standards in the standards and in the extracts of river waters and in sediments are shown in Table SM3 (Supplementary Material). Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 66 3.2.8 Validation The quality and consistency of the analytical results was check in method validation for the twenty-seven pharmaceuticals under study. SPE and QuEChERS were the methodologies used for the extraction of the compounds in river waters and in sediments and UHPLC-MS/MS was used for the analysis. Linearity, precision, matrix effect (ME), method detection limit (MDL), method quantification limit (MQL), and recovery were the parameters analyzed for both matrices. 3.2.9 Statistical analysis Descriptive statistics was used to summarize the results of pharmaceutical and physicochemical contamination in water and sediments samples. Pairwise correlation coefficients between the variables in the river water and sediments were done (Pearson correlation coefficient). Principal Component Analysis (PCA) was performed using XLSTAT Version 2019.3.1 to explore possible relationships between the levels of pharmaceuticals in water and sediment (both layers) with the water and sediments properties. Statistical significance was defined at p < 0.05. Of the twenty-seven pharmaceuticals, only the detected were considered in this analysis. When applicable, values were assumed to be equal to half of the limit of detection/quantification. 3.2.10 Risk assessment The protection of the aquatic ecosystem from pharmaceuticals pollution is utmost important and its impact on the environment should be evaluated. Risk assessment was determined following the work published by Paíga et al. (2019). 3.3 Results and discussion 3.3.1 Parameters The physical and chemical parameters were analyzed in river waters and in sediments from Douro and Leça rivers (Table SM5 and SM6, Supplementary Material). According to the particle size distribution results, all the sediments presented a sandy texture. The analysis for E.C., DO, chloride, nitrate and sulphate show the same profile in the samples WsD3, WsL1, and WsL2, standing out as the points with the lowest values for these parameters. Chapter 3 67 Figure 3.2 presents the levels of elements content in the sediments and river waters. The concentration levels and the pattern of distribution of elements in both matrices and different sampling points were different. The element quantity levels found in sediments were typically higher than in the corresponding river waters. The bottom sediment sample SsD3 present a higher concentration of almost all the analyzed elements. The main highlights to the river water were the sodium (64.85-67.65 mg/L in Leça and 69.62-4,863 mg/L in Douro), chloride (76-79 mg/L in Leça and 63-11 552 mg/L in Douro) and sulphate (38-71 mg/L in Leça and 51- 1,548 mg/L in Douro) which, together with the electrical conductivity, show an increase on the Douro River as it approaches the sea. This salinity increment in the river course near to ocean is coincident with other studies (Liu et al., 2019). This characteristic shows the slightest influence that the sea presents on the Leça river, since its mouth is located at Port of Leixões basin. Figure 3.2 - Element concentration in river water and sediment samples (Si in g/Kg). Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 74 highest detection frequency was observed for fluoxetine with 83.3% followed by carbamazepine with 66.7%. In Douro, only carbamazepine and fluoxetine were detected, carbamazepine with values lower than MDL in two samples and fluoxetine with a similar concentration (between 1.87 to 1.94 ng/L) in three of the four collected samples. In Leça, thirteen pharmaceuticals (azithromycin, ciprofloxacin, clarithromycin, moxifloxacin, ofloxacin, trimethoprim, carbamazepine, citalopram, diazepam, fluoxetine, sertraline, trazodone, and venlafaxine) were detected. Concentration ranged from <MDL (trazodone) to 2,819 ng/L (azithromycin) in WsL1 and between <MDL (citalopram and trazodone) to 235 ng/L (venlafaxine) in WsL2. The highest concentration was achieved for azithromycin in µg/L levels in sample WsL1 for antibiotics and carbamazepine and venlafaxine for psychiatric drugs in ng/L levels. For all pharmaceuticals higher levels were achieved in sample WsL1 when compared with sample WsL2. This can be explained by the location of the sampling point. Sample WsL1 was collected near the WWTP effluent discharge. The greatest difference between samples WsL1 and WsL2 was observed for azithromycin since the concentration found in sample WsL1 is approximately 12 times higher than the concentration found in sample WsL2. Referring to the Douro water sampling points and to the detected compounds, a slight reduction of concentrations along the river can be observed, except for point WsD3, in which no compound was detected. The presence of a water discharge point (of unknown origin) between the point WsD2 and WsD3 may have promoted the dilution of the pollutants at this point. The potential environmental risk posed by the detected pharmaceuticals in the Douro and Leça rivers was assessed in three different trophic levels (fish, Daphnia magna, and algae). Azithromycin, ciprofloxacin, clarithromycin, ofloxacin, trimethoprim, carbamazepine, citalopram, fluoxetine, sertraline, trazodone, and venlafaxine were the pharmaceuticals with measured concentrations in Leça river and carbamazepine and fluoxetine in Douro River. The risk quotient was calculated using the highest concentration detected in both rivers (Table 1). The results are presented in Figure SM3 (Supplementary Material). Carbamazepine exceeds the threshold value of one, showing a potential risk to algae and venlafaxine is too close to reach the environment potential risk also to algae, in Leça river. Chapter 3 75 3.3.3.2 Sediments Many pollutants, originally introduced into water, have affinity for sediments particles due to their hydrophobic/lipophilic properties (Perelo, 2010; Reid et al., 2000). Therefore, our study was extended to the extraction and analysis of pharmaceuticals in sediments and six sampling points were selected along Douro and Leça for collect sediments samples. Then, two samples were collected at each sampling point. One sample was collected at the top (0-2 cm depth) and the second sample was collected at the bottom (2-10 cm) zone (see section 2.6). Concentrations were compared not only between the different sampling points but also between different depths of the same sampling point (top, bottom) (Table 3.2). Table 3.2 - Concentration of detected antibiotics and psychiatric drugs expressed ng/g for sediment samples and relative standard deviation (%RSD). Pharmaceuticals organized by alphabetic order and by its therapeutic group. Detention frequency was calculated taking into account for all samples. Conc-Concentration; n.d.- Not detected; MDL-Method detection limit; RSD- Relative Standard Deviation Two antibiotics (azithromycin and sulfamethoxypyridazine) and six psychiatric drugs (carbamazepine, citalopram, fluoxetine, sertraline, trazodone, and venlafaxine) were detected in the sediments. Concentrations were below the MDL with exception of venlafaxine (0.251 ng/g) for sediments collected in Douro and from <MDL (carbamazepine and trazodone) to 43.2 ng/g (azithromycin) in Leça sediments. Fluoxetine had the highest detection frequency. Only one antibiotic was detected either in the Douro sediments (sulfamethoxypyridazine, <MDL, SsD4 top) either in the Leça sediments (azithromycin, 43.2 ng/g, SsL1 top). When top and bottom zones were compared, it must be taken into account that soils with sandy texture do not have sufficient organic matter that slows the migration of pharmaceuticals, increasing the risk of its infiltration through the soil (Fehsenfeld, 2015). In Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 76 the present study, sulfamethoxypyridazine, citalopram, fluoxetine, and trazodone detected in Douro sediments had insignificant differences between the top and the bottom zones due to the lower levels (<MDL) found. In the other hand, venlafaxine in sample SsD1 was not detected at the top and detected at the bottom zone, showing its migration through the SsD1 sediment sample. In Leça, results revealed lower concentrations in the bottom when compared with the top zone for azithromycin, citalopram, fluoxetine, sertraline, and venlafaxine. The detection of these pharmaceuticals in bottom zone also demonstrates the migration of the pharmaceuticals through sediment. For the authors, the higher concentration at the top zone, could be an evidence of recent contamination. For carbamazepine and trazodone, similar concentration in the two zones with values below the MDL was observed. 3.3.4 Statistical analysis Of the twenty-seven pharmaceuticals, only the detected were considered in PCA. In the general analysis, performed with all the water and sediment samples of both studied rivers, PCA expresses 42.46% of total variance, highlighting the opposition among the water samples WsL1 and WsL2, both from Leça River, with WsD4, from Douro River. SsD3 and SsD2 bottom and SsL1 and SsD1 top, are characterizes by the higher element concentration (except for B and Na). WsL1 and WsL2 were the most contaminated water samples and most associated with NO2, NH4+, Cl- and Eh (Figure 3.5.A). According to the Pearson matrix (data not shown), no significant correlations among levels of pharmaceuticals in water and sediments samples and its characterization were found except for the NH4+ and the pharmaceuticals moxifloxacin and diazepam. Chapter 3 77 Figure 3.5 - PCA analysis of pharmaceutical levels and characteristics of the water and sediments samples from Douro and Leça River (A); From Leça River (B) and from Douro River (C and D) (AZM – Azithromycin; SMP – Sulfamethoxypyridazine; CIP - Ciprofloxacin; CTM – Clarithromycin; MXF – Moxifloxacin; OFL – Ofloxacin; TMP - Trimethoprim; CBZ – Carbamazepine; CIT- Citalopram; DZ – Diazepam; FLU – Fluoxetine; SER – Sertraline; TRZ – Trazodone; VEN – Venlafaxine; Temp – Temperature). Between the sediments and water samples characteristics, some of the significant correlations found were the OM with the Silt and Clay percentage, Pb and Ca with the PO43- concentration and Ptotal with C and N percentage, Cd and Polsen concentration. In the Leça River samples, the first two principal components explained 45.49% and 32.56% of the total variance, respectively. Significant differences can be observed in the distribution of pharmaceutical substances in both water and sediments. High levels of pharmaceutical were found in the sample WsL1, as well as higher values of EC, pH H2O, SO42- and temperature. It is, also, accentuated the opposition among the sediment samples SsL1 bottom and top. PO43-, Sand and S were more associated with the bottom layer and the trace elements Hg, Mg, As, Cd, Co, Cr, Ni, and Pb with the top layer, as well as sertraline (SER) higher Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 78 concentration. pH (KCl and CaCl2), Polsen and Ptotal, OM and the percentage of Silt and Gravel were, also, more associated with the top layer sample (Figure 3.5.B). In addition, the significant correlations among pharmaceuticals concentration in water and sediments, and its characterization were found among the trazodone (TRZ) concentration, the E.C., and the concentration of Si, Ca, K, Fe and Mn. Also, SO42- presents a significant correlation with the concentration of clarithromycin (CTM), carbamazepine (CBZ) and venlafaxine (VEN). A strong correlation also can be observed between concentration of carbamazepine (CBZ) and E.C. In Douro River, the first two principal components explained 27,21% and 15.37% of the total variance, respectively. The samples WsD1, WsD2 and WsD4 were strongly associated with higher carbamazepine (CBZ) and fluoxetine (FLU) concentrations, and the samples SsD2 top and bottom between each other and with the SsD4 bottom and SsD1 top. The samples SsD3 bottom showed opposition relatively to the samples SsD1 bottom, SsD3 and SsD4 top and SsD5 top and bottom, presenting higher concentration of, PO43-, Ptotal, Polsen, NO3-, NO2-, Hg, Ca, Mg, Na, Co, Pb and percentage of Water content, OM, C, N, S and Gravel (Figure 5.C). Including the third components explaining, in accumulated, 55,93% of the total variance, some of the associations were emphasized, as the strong relation between SsD2 bottom and top and between WsD2 and WsD4 (Figure 5.D). Also, the opposition between the samples WsD3, SsD5 top and bottom and SsD1 bottom and SsD2 bottom, and top is highlighted. In addition, no significant correlations among pharmaceuticals concentration in the Douro samples and its characteristics were found except between fluoxetine (FLU) and Si concentration and with EC. 3.3.5 Concentration found in literature Research was carried out in the literature considering the matrix, the pharmaceuticals included in our study, and the works published in the last decade. The measured concentration obtained in these studies is listed in Table SM10 (Supplementary Material) for river waters (ng/L) and sediments (ng/g). Several monitoring studies were found for river waters however few studies were published with results from the monitoring of pharmaceuticals in sediments and even less in sediments and river waters from the same site. Moreover, using the keywords “pharmaceuticals”, “sediments”, and “Portugal”, thirty works were achieved, although these studies were focus on issues such as biodegradation, remediation, and bioremediation. At our knowledge and for our set of pharmaceuticals, no works were founded in monitoring studies Chapter 3 79 in Portugal for sediments. The pharmaceuticals detected and its concentrations worldwide are showed in Table SM10 (Supplementary Material) and discussed in the following paragraphs. The highest concentrations found for antibiotics in the literature stand out for sulfonamide antibiotics trimethoprim (13 600 ng/L) in Laizhou Bayin in China (R. Zhang et al., 2012) and for sulfamethazine (1 090 ng/L) and sulfamethoxazole (5 320 ng/L) in Msunduzi River in South Africa (Matongo et al., 2015) in river water and for fluoroquinolone ciprofloxacin (1 290 ng/g) in Hai River (L. J. Zhou et al., 2011) in sediments. The µg/L and µg/g levels found in these studies could be either an indication that point source pollution occurred in the studied area at the time of sampling as also a result of constant and aggressive pollution. For the remain studies, the concentration of antibiotics ranged from <MDL to ng/L or ng/g levels. Prulifloxacin and moxifloxacin were not detected in any work. In Pearl, Ba, and Huangpu rivers in China and Msunduzi river in South Africa, pharmaceuticals were assessed in both matrices (S. Li et al., 2018; Liang et al., 2013; Matongo et al., 2015) but more antibiotics were detected in Pearl river (S. Li et al., 2018; Liang et al., 2013). These studies were compared between themselves for each pharmaceutical and generally, when a concentration is higher in the river water is also higher in its sediments or vice versa. However, this pattern was not always verified. The unexpected profile can result in a point-source pollution that occurs in the area as also must be consider the migration of the pollutants through soils. Our results, in river water were in accordance with the results found in Pearl River but with lower concentration for trimethoprim (WsL1 sample). The two antibiotics found in our study for sediments collected in Douro and Leça rivers were not a target of studies in the literature. In Portugal, pharmaceuticals were analyzed in thirteen rivers (Antuã, Arade, Ave, Cértima, Guadiana, Leça, Lis, Mondego, Tagus, Tâmega, Trancão, Tua, and Xarrama rivers) but no studies were performed in sediments. Concentrations of antibiotics found in Leça River were higher than the concentrations found in the other studies conducted in Portugal. For psychiatric drugs, all studies were carried out in river waters except for the study performed in South Africa in which both matrices were assessed. Matongo et al. in 2015 found for carbamazepine concentration between 130 to 3 240 ng/L in Msunduzi River and between <0.535 to 6.07 ng/g in sediments (Matongo et al., 2015). The highest concentration of psychiatric drugs (carbamazepine) was found in the study of Matongo et al. The levels found in our study were in accordance with the remain results in Portugal, however for sample WsL1 (Leça river) venlafaxine have higher concentration. Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 80 3.4 Conclusions In multiresidue method, good results for all compounds are not always found in one single extraction, namely: sulfonamides and fluoroquinolones. The differences of the physicochemical properties of these compounds and the extraction conditions used often do not allow higher recoveries for all target analytes. Optimizing single extraction for each compound is not acceptable since the costs, extraction time, solvents, and the produced wastes would be increased, and the principles of Green Chemistry are set aside. In our study, the adopted extraction procedures, SPE and QuEChERS methodologies, yielded good recoveries for most of the study pharmaceuticals. River waters and sediments (top and bottom) were collected along Douro and Leça rivers. All samples were characterized, and pharmaceuticals were analyzed in both matrices. Higher concentration was found in samples (river water and sediments) collected in Leça when compared with samples collected in Douro. In river water, only two psychiatric drugs were found (carbamazepine and fluoxetine). In the other hand, thirteen (six antibiotics and sevens psychiatric drugs) of the twenty-seven pharmaceuticals were detected in Leça river waters. It must be highlighted the highest concentration found for azithromycin in µg/L level. Sampling point WsL1 is located near to the WWTP effluent discharge, this can justify the values found. Carbamazepine and venlafaxine showed a potential risk to algae in Leça river. In the case of sediments, sulfamethoxypyridazine was found in Douro sediment and azithromycin in Leça sediment. Psychiatric drugs were found in the collected samples: four in Douro sediment) and six in Leça sediments. The concentration found in the top and at the bottom in each sample were compared: in Douro sediment venlafaxine had a higher concentration in the bottom zone which could be correlated with the migration of the pharmaceutical through the soil. In Leça sediments higher concentration was found in top zone, which could be attributed to recent pollution in the studied area. Comparing the works reported in literature there is no pattern of the concentration found in both matrices due to the point-source pollutions in the river water and migration of the pollutants in the sediments. From the results obtained in our study, more monitoring studies need to be performed in all courses of the rivers. Source of pollution need to be identified; thus samples need to be collected in the rural area, in the industrialized area, and near to the harbor. As few studies were found in literature in sediments as also embracing together river waters and sediments, the contribution of this article is important for further monitoring Chapter 3 81 studies. Pharmaceutical pollution not only reaches the watercourses, because also sediments, soil, and groundwater are affected. CRediT authorship contribution statement Maria Joao Fernandes: ~ Writing - original draft, Investigation, Data curation. Paula Paíga: Writing - original draft, Investigation, Validation, Data curation. Ana Silva: Writing - original draft, Resources. Carmen Perez Llaguno: Investigation, Validation. Manuela Carvalho: Formal analysis, Resources, Writing - review & editing. Felipe Macías Vazquez: Conceptualization, Formal analysis, Resources, Writing - review & editing, Supervision. Cristina Delerue-Matos: Conceptualization, Formal analysis, Resources, Writing - review & editing, Project administration, Supervision. Acknowledgment This work was also supported by UID/QUI/50006/2019 with funding from FCT/MCTES through national funds. The authors would like to thank also the EU and FCT / UEFISCDI /FORMAS for funding, in the frame of the collaborative international consortium REWATER financed under the ERA-NET Cofund WaterWorks2015 Call. This ERA-NET is an integral part of the 2016 Joint Activities developed by the Water Challenges for a Changing World Joint Programme Initiative (Water JPI) Appendix A. Supplementary data Supplementary data to this article can be found online at https://doi.org/10.1016/j.chemosphere.2019.124729. Chapter 3 82 Appendix A. Supplementary data Table SM1. Pharmaceuticals, metabolites, isotopically-labelled internal standards, Chemical Abstracts Service (CAS), molecular weight, formula, structure, solvent used, and supplier company. Pharmaceuticals and Isotopically-Labelled Internal Standards CAS*, ¥ Molecular Weight ¥ (g/mol) Formula and Structure¥ pka Solvent used for the preparation of the stock solution Supplier Company Antibiotics Azithromycin 83905-01-5 748.996 C38H72N2O12 Strongest acidic pKa: 12.43 Strongest basic pKa: 9.57 Methanol Sigma-Aldrich (Madrid, Spain) Ciprofloxacin 85721-33-1 331.347 C17H18FN3O3 5.76; 8.68 Milli-Q Water-10% acetic acid in Milli-Q water (1:1,v/v) Sigma-Aldrich (Madrid, Spain) Clarithromycin 81103-11-9 747.964 C38H69NO13 Strongest acidic pKa: 12.46 Strongest basic pKa: 8.38 Methanol Sigma-Aldrich (Madrid, Spain) Enrofloxacin 93106-60-6 359.401 C19H22FN3O3 5.69: 6.68 Milli-Q Water-10% acetic acid in Milli-Q water (1:1, v/v) Sigma-Aldrich (Madrid, Spain) Erythromycin 643-22-1 733.937 C37H67NO13 Strongest acidic pKa: 12.45 Strongest basic pKa: 8.38 Methanol Sigma-Aldrich (Madrid, Spain) Chapter 3 83 Lomefloxacin 98079-51-7 351.354 C17H19F2N3O3 Strongest acidic pKa: 5.45 Strongest basic pKa: 8.7 Milli-Q Water-10% acetic acid in Milli-Q water (1:1,v/v) Moxifloxacin 151096-09-2 401.438 C21H24FN3O4 Strongest acidic pKa: 5.49 Strongest basic pKa: 9.42 Milli-Q Water-10% acetic acid in Milli-Q water (1:1,v/v) Norfloxacin 70458-96-7 319.33 C16H18FN3O3 Strongest acidic pKa: 5.58 Strongest basic pKa: 8.68 Milli-Q Water-10% acetic acid in Milli-Q water (1:1,v/v) Sigma-Aldrich (Madrid, Spain) Ofloxacin 82419-36-1 361.373 C18H20FN3O4 5.45; 6.20 Milli-Q Water-10% acetic acid in Milli-Q water (1:1,v/v) Sigma-Aldrich (Madrid, Spain) Prulifloxacin 123447-62-1 461.464 C21H20FN3O6S Strongest acidic pKa: 5.93 Strongest basic pKa: 5.27 Milli-Q Water-10% acetic acid in Milli-Q water (1:1,v/v) Toronto Research Chemicals Inc. (Ontario, Canada) Sulfadiazine 68-35-9 250.276 C10H10N4O2S 2.01; 6.99 Methanol Sigma-Aldrich (Madrid, Spain) Sulfadimethoxine 122-11-2 310.328 C12H14N4O4S 1.95; 6.91 Methanol Sigma-Aldrich (Madrid, Spain) Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 90 Table SM4. Retention time and ion ratio for antibiotic, psychiatric drugs, and isotopically-labelled internal standards. Pharmaceuticals tr (min) Ion Ratio (%) tr average (min) RSD (%) n -20% Ion Ratio Average (%) +20% RSD (%) n Antibiotics Azithromycin 2.317 0.234 30 1.36 1.71 2.05 4.37 18 Ciprofloxacin 2.265 0.321 30 0.942 1.18 1.41 3.30 13 Clarithromycin 2.751 0.140 30 0.968 1.21 1.45 1.87 20 Enrofloxacin 2.302 0.313 30 4.50 5.62 6.75 3.74 17 Erythromycin 2.606 0.100 30 3.29 4.11 4.93 2.37 16 Lomefloxacin 2.279 0.236 30 0.814 1.02 1.22 4.90 17 Moxifloxacin 2.403 0.356 30 0.897 1.12 1.35 3.44 27 Norfloxacin 2.250 0.219 30 0.964 1.21 1.45 3.80 26 Ofloxacin 2.249 0.326 30 1.22 1.52 1.83 2.92 25 Prulifloxacin 2.701 0.165 30 0.993 1.24 1.49 4.05 27 Sulfadiazine 2.435 0.307 30 1.13 1.42 1.70 2.55 28 Sulfadimethoxine 2.989 0.0946 30 1.60 2.00 2.40 1.92 27 Sulfamethazine 2.629 0.214 30 1.00 1.25 1.50 2.90 22 Sulfamethoxazole 2.856 0.114 30 0.955 1.19 1.43 2.47 28 Sulfamethoxypyridazine 2.634 0.110 30 1.53 1.92 2.30 3.10 23 Sulfapyridine 2.482 0.164 30 0.777 0.97 1.16 2.54 23 Trimethoprim 2.217 0.199 30 2.18 2.72 3.27 3.85 18 Psychiatric drugs Carbamazepine 3.095 0.0843 30 3.81 4.76 5.72 2.86 28 Citalopram 2.631 0.118 30 27.9 34.8 41.8 4.94 21 Diazepam 3.529 0.110 30 1.01 1.26 1.51 2.51 25 10,11-Epoxycarbamazepine 2.869 0.108 30 1.12 1.40 1.68 2.99 26 Fluoxetine 2.789 0.0991 30 2.22 2.78 3.33 2.85 25 Norfluoxetine 2.763 0.389 30 2.60 3.25 3.90 2.03 24 Paroxetine 2.694 0.0770 30 0.833 1.04 1.25 3.60 28 Sertraline 2.811 0.160 30 1.16 1.45 1.74 2.35 24 Trazodone 2.501 0.0702 30 0.923 1.15 1.39 4.07 25 Venlafaxine 2.462 0.116 30 0.888 1.11 1.33 2.36 27 Isotopically-labelled internal standards Azithromycin-d3 2.315 0.152 30 Carbamazepine-d10 3.080 0.101 30 Ciprofloxacin-d8 2.262 0.132 30 Diazepam-d5 3.515 0.0500 30 Fluoxetine-d5 2.785 0.0974 30 Sulfamethoxazole-d4 2.851 0.0664 30 Venlafaxine-d6 2.462 0.0878 30 Pharmaceuticals organized by alphabetic order and by its therapeutic group. Chapter 3 91 Table SM5. Analyzed parameters in sediment samples from Douro and Leça rivers. Parameters SsD1 top SsD1 bottom SsD2 top SsD2 bottom SsD3 top SsD3 bottom SsD4 top SsD4 bottom SsD5 top SsD5 bottom SsL1 top SsL1 bottom pH H2O 8.10 7.55 8.06 7.15 8.31 7.21 7.85 7.58 7.62 7.56 6.74 6.36 pH KCl 7.90 7.15 8.07 7.00 8.55 6.87 7.89 7.55 7.91 7.18 6.42 5.91 pH CaCl2 7.32 6.97 7.45 6.96 n.a. 6.79 7.31 n.a. 7.08 6.97 6.07 6.06 C.E. (mS cm-1) 3.01 1.49 5.72 7.29 n.a. 4.21 5.70 n.a. 2.57 1.50 0.44 0.36 Water Content (%) 9.4 14.9 33.7 39.0 n.a. 18.8 21.2 n.a. 7.0 4.4 74.4 22.25 OM (%) 2.03 1.02 3.12 5.65 n.a. 2.21 1.30 n.a. 1.45 0.29 1.88 1.00 SO42- (g L-1) n.d. 0.22 n.d. 1.89 n.a. n.d. n.d. n.a. 0.29 0.23 n.d. 0.28 PO43- (mg L-1) n.d. 0.06 0.18 n.d. n.a. 1.22 n.d. n.a. n.d. 0.01 n.d. 0.01 Ptotal (mg/Kg) 395.50 218.19 n.a. 432.72 251.37 614.24 371.38 345.31 185.54 156.30 1,115.25 n.d. Polsen (mg/Kg) 30.63 4.55 n.a. 13.32 6.96 40.18 16.79 15.92 5.31 3.40 54.71 n.d. C (%) 0.99 0.26 n.a. 0.57 0.28 2.10 0.44 0.59 0.28 0.21 2.30 n.d. N (%) 0.17 0.03 n.a. 0.04 0.03 0.14 0.04 0.08 0.02 0.03 0.27 n.d. S (%) 0.051 0.004 0.320 0.053 0.020 0.168 0.055 0.060 0.014 0.01 0.137 0.195 Gravel (%) 65.37 48.71 9.61 7.21 n.a. 62.74 22.96 n.a. 12.05 34.62 30.20 4.40 Sand (%) 32.61 51.03 81.92 80.51 n.a. 36.93 76.02 n.a. 87.95 65.38 69.32 95.60 Silt (%) 1.97 0.26 8.06 11.30 n.a. 0.33 1.02 n.a. 0.00 b 0.00 b 0.48 0.00b Clay (%) 0.05 0.00a 0.41 0.98 n.a. 0.00 a 0.00 a n.a. 0.00 a 0.00 a 0.00 a 0.00 a Texture Sandy Sandy Sandy Sandy n.a. Sandy Sandy - Sandy Sandy Sandy Sandy n.a. – not analysed a – the result of the sum of all the granulometric fractions under 0.002 mm was 0.00. b – the result of the sum of all the granulometric fractions between 0.002-0.02 mm was 0.00. Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 92 Table SM6. Analysed parameters in water samples from Douro and Leça rivers. Parameters WsD 1 WsD 2 WsD 3 WsD 4 WsL 1 WsL 2 Temperature (ºC) 19.8 18.5 19.4 19.0 18.9 14.6 pH 7.24 6.09 7.38 7.20 7.15 7.05 E.C. (μS/cm) 11,490 15,770 578 20,600 932 818 Eh (mV) 456 403 383 360 438 483 DO (mg/L) 11.0 11.6 n.a. 11.8 n.a. n.a. F- (mg/L) 0.39 0.54 1.01 0.67 0.09 0.10 Cl- (mg/L) 5,658 8,713 63 11,552 76 79 SO42- (mg/L) 705 1,040 51 1,548 71 38 NH4+ (mg/L) < 0.03 < 0.03 1.16 0.22 6.70 22.21 NO3- (mg/L) 10 12 1 38 n.a. 3 NO2- (mg/L) 0.10 0.64 0.13 1.65 2.62 3.07 n.a. – not analysed Chapter 3 93 Section 1. Method validation Calibration curve Linearity was studied using standards prepared in the solvent, in river waters, and in sediments extracts. Thus, for all pharmaceuticals, ten concentration levels for calibration curve in solvent between 5.0 and 1,000 µg/L and six concentration levels for a matrix-matched calibration curve between 5.0 and 500 µg/L were used. Calibration curves were made in both MRM transitions. Three replications (with an injection volume of 5 μL) were made for each standard solution to check the repeatability at each concentration level. Method Limits of detection (MDL) and quantification (MQL) MDL and MQL were determined as the minimum amount detectable of the analyte with a signal-to-noise ratio of 3 and 10, respectively. The limits were determined in river waters and in sediments but only if the pharmaceutical was detected in these samples, otherwise limits were determined in the matrix-matched calibration curve. Repeatability Repeatability analysis expressed as the relative standard deviation (RSD (%)). Three concentration of standards (100, 250, 500 µg/L) were repeated six-time sequential and repeated along the batch (n=6). Matrix effect The influence of the ME was evaluated comparing one standard prepared in solvent and one standard prepared in the matrix (Jelić et al., 2012). For each compound recovery was calculated by comparing its area in the samples spiked prior to SPE extraction (pre-spiked sample) with the area in the samples spiked after SPE extraction (post-spiked sample) (Paíga and Delerue-Matos, 2016). Thus, for river waters and sediments, a blank and fortified sample were carried out for the pre- and post-spiked samples. A positive value of ME is an indicative of ion enhancement signal and a negative value represents an ion suppression signal (Jelić et al., 2012). References Jelić, A., Gros, M., Petrovic, M., Ginebreda, A., Barceló, D., 2012. Occurrence and Elimination of Pharmaceuticals During Conventional Wastewater Treatment. Emerg. Prior. Pollut. Rivers. Handb. Environ. Chem. 19, 1–24. https://doi.org/10.1007/978-3-642-25722-3 Paíga, P., Delerue-Matos, C., 2016. A throughput method using the quick easy cheap effective rugged safe method for the quantification of ibuprofen and its main metabolites in soils. J. Sep. Sci. 39, 3436–3444. https://doi.org/10.1002/jssc.201600412 Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 94 Table SM7. Method detection and method quantification limits (MDL, MQL) in river water (ng/L) and sediment samples (ng/g). Pharmaceuticals River water Decision 2018/840 of 5 June 2018* MDL (ng/L) Sediment** MDL (ng/L) MQL (ng/L) MDL (ng/g) MQL (ng/g) Antibiotics n.a. * Azithromycin 3.98 13.3 19 0.00200 0.00667 * Ciprofloxacin 41.1 137 89 1.04 3.46 * Clarithromycin 0.0750 0.225 19 0.00200 0.00667 Enrofloxacin 1.55 5.18 n.a. 1.31 4.38 * Erythromycin 9.20 30.7 19 0.00400 0.0150 Lomefloxacin 37.8 126 n.a. 5.43 18.1 Moxifloxacin 39.6 132 n.a. 1.16 3.85 Norfloxacin 23.5 78.3 n.a. 3.39 11.3 Ofloxacin 0.450 1.53 n.a. 0.0170 0.0560 Prulifloxacin 12.1 40.4 n.a. 0.294 0.979 Sulfadiazine 7.50 25.0 n.a. 0.0430 0.144 Sulfadimethoxine 65.2 217 n.a. 0.0300 0.0990 Sulfamethazine 2.35 7.80 n.a. 0.114 0.379 Sulfamethoxazole 0.150 0.475 n.a. 0.225 0.752 Sulfamethoxypyridazine 55.5 185 n.a. 0.0490 0.163 Sulfapyridine 9.33 31.1 n.a. 0.0660 0.219 Trimethoprim 9.45 31.5 n.a. 0.0810 0.272 Psychiatricdrugs Carbamazepine 1.05 3.48 n.a. 0.00643 0.0257 Citalopram 0.500 1.63 n.a. 0.00429 0.0143 Diazepam 3.70 12.35 n.a. 0.00400 0.0133 10,11-Epoxycarbamazepine 2.28 7.60 n.a. 0.00900 0.0300 Fluoxetine 0.225 0.750 n.a. 0.00214 0.00710 Norfluoxetine 8.25 27.5 n.a. 0.189 0.630 Paroxetine 0.0500 0.150 n.a. 0.369 1.23 Sertraline 0.125 0.425 n.a. 0.00214 0.00710 Trazodone 0.850 2.85 n.a. 0.00900 0.0300 Venlafaxine 0.575 1.93 n.a. 0.0364 0.120 Pharmaceuticals organized by alphabetic order and by its therapeutic group. *Pharmaceuticals with maximum acceptable method detection limit (ng/l) in the Commission Implementing Decision (EU) 2018/840 of 5 June 2018 (Official Journal of the European Union, L 141/9. COMMISSION IMPLEMENTING DECISION (EU) 2018/840 of 5 June 2018 establishing a watch list of substances for Union-wide monitoring in the field of water policy pursuant to Directive 2008/105/EC of the European Parliament and of the Council and repealing Commission Implementing Decision (EU) 2015/495. Available at: https://eur-lex.europa.eu/legalcontent/EN/TXT/PDF/?uri=CELEX:32018D0840&from=EN, Accessed data: November 2018) n.a. - No Limit was applicable in the water samples ** - No Limit was applicable in the sediments samples Chapter 3 95 Table SM8. Recoveries (%) at three levels of fortification for river sample matrix. Pharmaceuticals Level I Level II Level III Average recovery* Recovery (%) RSD (%) Recovery (%) RSD (%) Recovery (%) RSD (%) Antibiotics Azithromycin 66.5 3.7 73.7 6.1 Note Note 70.1 Ciprofloxacin 102 5.1 95.6 6.7 96.5 2.7 98.0 Clarithromycin 74.0 2.0 67.0 4.1 70.5 1.1 70.5 Enrofloxacin 83.6 2.3 78.9 2.3 83.8 3.7 82.1 Erythromycin 82.3 4.6 78.7 4.1 80.8 6.7 80.6 Lomefloxacin 107.9 4.1 102 5.4 94.6 3.4 101.5 Moxifloxacin 87.4 1.2 88.8 4.7 79.6 1.4 85.3 Norfloxacin 92.4 4.3 96.2 4.6 98.8 5.3 95.8 Ofloxacin 71.1 2.7 70.3 3.7 64.6 2.5 68.7 Prulifloxacin 86.8 2.1 94.3 0.57 102 1.8 94.3 Sulfadiazine 52.1 3.4 49.1 5.7 56.5 3.1 52.6 Sulfadimethoxine 56.6 5.8 61.5 3.8 55.2 4.1 57.8 Sulfamethazine 51.2 3.9 52.4 6.6 54.7 2.3 52.8 Sulfamethoxazole 47.8 1.8 50.5 3.2 53.4 5.3 50.6 Sulfamethoxypiridazine 58.1 4.1 53.7 2.0 51.4 3.4 54.4 Sulfapyridine 55.5 2.1 52.6 4.1 52.9 2.7 53.7 Trimethoprim 91.3 2.1 95.6 3.8 97.5 3.5 94.8 Psychiatric drugs Carbamazepine 103 3.1 95.1 2.9 98.6 1.8 98.9 Citalopram 89.5 9.3 89.5 2.7 85.5 2.8 88.2 Diazepam 97.0 3.9 94.4 1.1 94.8 6.6 95.4 10,11-Epoxycarbamazepine 63.7 2.6 64.9 4.4 69.1 0.59 65.9 Fluoxetine 90.5 2.5 88.3 1.6 85.1 1.6 88.0 Norfluoxetine 80.6 4.3 77.8 3.8 81.4 2.5 79.9 Paroxetine 83.5 4.8 84.2 1.2 85.8 5.4 84.5 Sertraline 85.7 1.6 91.3 1.5 86.1 1.9 87.7 Trazodone 89.1 1.7 93.2 3.7 81.7 2.5 88.0 Venlafaxine 91.1 5.4 98.2 3.6 93.4 5.4 94.2 Pharmaceuticals organized by alphabetic order and by its therapeutic group. * Average recovery embracing the three fortified levels. Note: in spiking level III azithromycin was not detect. Some problem must have occurred during the extraction and this level was discharged. Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 96 Table SM9. Recoveries (%) at three levels of fortification for sediment sample matrix. Pharmaceuticals Level I Level II Level III Average recovery* Recovery (%) RSD (%) Recovery (%) RSD (%) Recovery (%) RSD (%) Antibiotics Azithromycin 66.9 2.8 74.4 4.5 87.8 2.4 76.4 Ciprofloxacin 2.15 49 0.89 41 0.340 33 1.13 Clarithromycin 76.9 2.0 80.9 5.6 83.7 7.6 80.5 Enrofloxacin 10.2 6.3 13.1 34 7.39 49 10.2 Erythromycin 86.4 0.66 63.5 7.4 64.3 12 71.4 Lomefloxacin 3.16 35 5.8 58 2.52 60 3.82 Moxifloxacin 3.11 65 3.3 40 1.92 96 2.76 Norfloxacin 2.86 53 0.75 48 0.512 16 1.37 Ofloxacin 2.29 37 1.9 61 1.20 65 1.80 Prulifloxacin 16.0 20 18.8 3.5 18.2 12 17.7 Sulfadiazine 13.3 18 17.2 29 15.9 1.0 15.4 Sulfadimethoxine 29.1 17 31.5 25 30.1 19 30.2 Sulfamethazine 18.7 8.8 26.2 4.1 29.5 4.3 24.8 Sulfamethoxazole 28.1 21 29.5 22 31.2 19 29.6 Sulfamethoxypiridazine 13.9 22 21.4 28 21.2 4.7 18.8 Sulfapyridine 18.0 11 27.8 31 24.9 12 23.6 Trimethoprim 74.8 7.0 92.8 7.7 78.2 6.2 81.9 Psychiatric drugs Carbamazepine 91.4 6.1 103 3.6 91.9 5.1 95.6 Citalopram 71.0 7.4 74.3 4.6 68.6 3.0 71.3 Diazepam 100 6.1 108 5.0 106 5.0 104.3 10,11-Epoxycarbamazepine 73.9 2.9 81.9 6.8 73.2 2.6 76.3 Fluoxetine 68.7 2.6 75.7 1.2 67.2 6.7 70.5 Norfluoxetine 63.6 10 67.9 8.2 57.8 4.8 63.1 Paroxetine 56.8 3.3 64.2 1.9 54.8 2.9 58.6 Sertraline 46.6 11 50.7 3.8 44.2 6.0 47.2 Trazodone 59.9 10 66.6 6.4 62.7 6.0 63.1 Venlafaxine 70.4 6.0 75.9 3.6 72.4 5.0 72.9 Pharmaceuticals organized by alphabetic order and by its therapeutic group. * Average recovery embracing the three fortified levels. Chapter 3 97 Table SM10. Measured concentrations (ng/L) for the target analytes river water and sediment from the research conducted by the scientific community in the last years. Pharmaceuticals Continent River (Country) Water Concentration (ng/L) Sediment Concentration (ng/g) Reference Antibiotics Ciprofloxacin Asia China (Xiangjiang River) n.d. (Lin et al., 2018) Asia Pearl River Estuary (Zhuhai City, China) 24.6-365 0.09-104 (Li et al., 2018) Asia Ba River (Lantian County, Shannxi Province) 2.66-28.6 2.40-62.5 (Jia et al., 2018) Asia Yellow River (China) n.d.-32.8 (Zhou et al., 2011) Asia Hai River (China) 2.05 - 1,290 (Zhou et al., 2011) Asia Liao River (China) n.d.-28.7 (Zhou et al., 2011) Asia Huangpu River (Shangai, China) n.d. - <LOQ (9.33) (Jiang et al., 2011) Asia Huangpu River (Shanghai, China) n.d.-34.2 (Chen and Zhou, 2014) Asia Laizhou Bay-river discharge points (China) n.d. - 346 (Zhang et al., 2012) Asia Chao Phraya River (Thailand) 48 (max.) (Tewari et al., 2013) Europe Seine River (Paris, France) n.d. (Tamtam et al., 2008) Europe Tâmega River (Chaves, Portugal) n.d. (Pereira et al., 2017) Europe Tua River (Mirandela, Portugal) n.d. (Pereira et al., 2017) Europe Mondego River (Portugal) n.d. (Pereira et al., 2017) Europe Trancão River (Lisbon, Portugal) n.d. (Pereira et al., 2017) Europe Tagus River (Lisbon, Portugal) n.d. (Pereira et al., 2017) Europe Xarrama River (Évora, Portugal) n.d. (Pereira et al., 2017) Europe Guadiana River (Vila Real de Santo António, Portugal) n.d. (Pereira et al., 2017) Europe Lis River (Leiria, Portugal) n.d.-88.7 (Paíga et al., 2016) Enrofloxacin Asia China (Xiangjiang River) n.d. (Lin et al., 2018) Asia Huangpu River (Shangai, China) n.d. - <LOQ (11.34) (Jiang et al., 2011) Asia Laizhou Bay-river discharge points (China) n.d.-24.6 (Zhang et al., 2012) Asia Pearl River (China) n.d. 1.43* (Liang et al., 2013) Asia Huangpu River (Shanghai, China) n.d.-14.6 n.d.-8.9 (Chen and Zhou, 2014) Europe Seine River (Paris, France) n.d. (Tamtam et al., 2008) Europe Ave River (North of Portugal) n.d. (Barbosa et al., 2018) Europe Leça River (North of Portugal) n.d.-44.99 (Barbosa et al., 2018) Europe Antuã River (North of Portugal) n.d.-343.28 (Barbosa et al., 2018) Europe Cértima River (North-Central Portugal) n.d.-45.53 (Barbosa et al., 2018) Ofloxacin Asia China (Xiangjiang River) n.d.-23 (Lin et al., 2018) Asia Pearl River Estuary (Zhuhai City, China) 0.80–195 n.d.-157 (Li et al., 2018) Asia Qiantang River basin (China) 40 62 (Chen et al., 2012) Asia Yellow River (China) n.d.-124 (Zhou et al., 2011) Asia Hai River (China) 1.96-653 (Zhou et al., 2011) Asia Liao River (China) n.d.-50.5 (Zhou et al., 2011) Asia Laizhou Bay-river discharge points (China) n.d.-45.4 (Zhang et al., 2012) Asia Pearl River (China) 15.8* 13.7* (Liang et al., 2013) Asia Huangpu River (Shangai, China) <LOQ (8.34) (Jiang et al., 2011) Asia Huangpu River (Shanghai, China) n.d.-28.5 n.d.-12.4 (Chen and Zhou, 2014) Europe Seine River (Paris, France) n.d.-30 (Tamtam et al., 2008) Europe Ave River (North of Portugal) n.d. (Barbosa et al., 2018) Europe Leça River (North of Portugal) n.d. (Barbosa et al., 2018) Europe Antuã River (North of Portugal) n.d. (Barbosa et al., 2018) Europe Cértima River (North-Central Portugal) n.d. (Barbosa et al., 2018) Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 98 Norfloxacin Asia China (Xiangjiang River) n.d. (Lin et al., 2018) Asia Pearl River Estuary (Zhuhai City, China) 32.8-381 3.38-444 (Li et al., 2018) Asia Ba River (Lantian County, Shannxi Province) 5.33-122 6.04-680 (Jia et al., 2018) Asia Qiantang River basin (China) n.d.-500 (Chen et al., 2012) Asia Yellow River (China) n.d.-142 (Zhou et al., 2011) Asia Hai River (China) n.d.-5770 (Zhou et al., 2011) Asia Liao River (China) n.d.-177 (Zhou et al., 2011) Asia Laizhou Bay-river discharge points (China) n.d. - 572 (Zhang et al., 2012) Asia Pearl River (China) 136* 20.5* (Liang et al., 2013) Asia Huangpu River (Shangai, China) n.d. (Jiang et al., 2011) Asia Huangpu River (Shanghai, China) n.d.-0.2 (Chen and Zhou, 2014) Europe Seine River (Paris, France) n.d.-46 (Tamtam et al., 2008) Moxifloxacin Lomefloxacin Asia Yellow River (China) n.d. (Zhou et al., 2011) Asia Hai River (China) 1.67 (Zhou et al., 2011) Asia Liao River (China) n.d. (Zhou et al., 2011) Europe Seine River (Paris, France) n.d. (Tamtam et al., 2008) Prulifloxacin Trimethoprim Asia China (Xiangjiang River) n.d.-93 (Lin et al., 2018) Asia Pearl River Estuary (Zhuhai City, China) n.d.-492 n.d.-210 (Li et al., 2018) Asia Ba River (Lantian County, Shannxi Province, China) 11.3-106 0.79-31.4 (Jia et al., 2018) Asia Qiantang River basin (China) n.d. (Chen et al., 2012) Asia Yellow River (China) n.d.-2.86 (Zhou et al., 2011) Asia Hai River (China) n.d.-5.63 (Zhou et al., 2011) Asia Liao River (China) n.d.-9.84 (Zhou et al., 2011) Asia Brahmaputra River (Bangladesh) < 0.08-17.20 (Hossain et al., 2018) Asia Chao Phraya River (Thailand) 3* (Tewari et al., 2013) Asia Laizhou Bay-river discharge points (China) n.d.-13,600 (Zhang et al., 2012) Africa Msunduzi River, KwaZulu-Natal, South Africa n.d.-290 <MDL (0.246)-87.55 (Matongo et al., 2015) America Hudson River (New York) n.d.-350 (Cantwell et al., 2018) Europe Danube River (Serbia, Belgrade) n.d.-25 (Grujić et al., 2009) Europe Sava River (Serbia, Belgrade) n.d. (Grujić et al., 2009) Europe Tamis River (Serbia) 24 (Grujić et al., 2009) Europe Seine River (Paris, France) 12-27 (Tamtam et al., 2008) Europe River Foss (United Kingdom) n.d.-76 (Burns et al., 2018) Europe River Ouse (United Kingdom) n.d.-22.9 (Burns et al., 2018) Europe Lis River (Leiria, Portugal) n.d. (Paíga et al., 2016) Europe Ave River (North of Portugal) n.d. (Barbosa et al., 2018) Europe Leça River (North of Portugal) n.d.-15.63 (Barbosa et al., 2018) Europe Antuã River (North of Portugal) n.d.-64.14 (Barbosa et al., 2018) Europe Cértima River (North-Central Portugal) n.d.-12.68 (Barbosa et al., 2018) Sulfamethoxypyridazine Asia Luan River (North China) 0.16-3.1 (Li et al., 2014) Europe Ebro River (Northeast Spain) 0.6-18.1 (García-Galán et al., 2011) Sulfapyridine Asia Pearl River Estuary (Zhuhai City, China) n.d.-58.3 n.d.-126 (Li et al., 2018) Asia Yellow River (China) n.d. (Zhou et al., 2011) Asia Hai River (China) n.d. (Zhou et al., 2011) Asia Liao River (China) n.d. (Zhou et al., 2011) Asia Huangpu River (Shanghai, China) 2.54-16.6 (Jiang et al., 2011) Asia Huangpu River (Shanghai, China) n.d.-103.1 n.d.-6.6 (Chen and Zhou, 2014) Chapter 3 99 Asia Luan River (North China) <0.1-3.4 (Li et al., 2014) Europe Ebro River (Northeast Spain) 0.1-42.5 (García-Galán et al., 2011) Europe Lis River (Leiria, Portugal) n.d. (Paíga et al., 2016) Sulfamethazine Asia Xiangjiang River (China) n.d.-60 (Lin et al., 2018) Asia Pearl River Estuary (Zhuhai City, China) n.d.-226 n.d.-72.3 (Li et al., 2018) Asia Ba River (Lantian County, Shannxi Province) 8.64-464 7.80-690 (Jia et al., 2018) Asia Yellow River (China) n.d. (Zhou et al., 2011) Asia Hai River (China) n.d.-5.69 (Zhou et al., 2011) Asia Liao River (China) n.d. (Zhou et al., 2011) Asia Brahmaputra River (Bangladesh) < 0.01-11.35 (Hossain et al., 2018) Asia Luan River (North China) 0.21-3.7 (Li et al., 2014) Asia Huangpu River (Shangai, China) 5.45-313.44 (Jiang et al., 2011) Asia Huangpu River (Shanghai, China) 19.9-389.4 0.2-2.7 (Chen and Zhou, 2014) Asia Laizhou Bay-river discharge points (China) <LOQ - 108 (Zhang et al., 2012) Africa Msunduzi River, KwaZulu-Natal, South Africa n.d.-1,090 n.d. (Matongo et al., 2015) Asia Pearl River (China) 218* 3.24* (Liang et al., 2013) Asia Chao Phraya River (Thailand) 5* (Tewari et al., 2013) Europe Ebro River (Northeast Spain) 2.5-65.2 (García-Galán et al., 2011) Europe Seine River (Paris, France) n.d. (Tamtam et al., 2008) Europe Lis River (Leiria, Portugal) n.d.-123 (Paíga et al., 2016) Europe Mess River (Luxembourg) 19 (max) (Meyer et al., 2011) Sulfadimethoxine Asia Brahmaputra River (Bangladesh) n.d. (Hossain et al., 2018) Asia Luan River (North China) 0.95-3.56 (Li et al., 2014) Europe Ebro River (Northeast Spain) 1-23.1 (García-Galán et al., 2011) Europe Mess River (Luxembourg) n.d. (Meyer et al., 2011) Sulfadiazine Asia China (Xiangjiang River) n.d.-68 (Lin et al., 2018) Asia Pearl River Estuary (Zhuhai City, China) n.d.-50.7 n.d.-198 (Li et al., 2018) Asia Ba River (Lantian County, Shannxi Province) 1.69-36.8 0.59-7.10 (Jia et al., 2018) Asia China, Hanjiang River (through Hubei and Shaanxi) 1.9-6.3 0.35-0.54 (Hu et al., 2018) Asia Yellow River (China) n.d.-22.0 (Zhou et al., 2011) Asia Hai River (China) n.d.-1.71 (Zhou et al., 2011) Asia Liao River (China) n.d.-11 (Zhou et al., 2011) Asia Huangpu River (Shangai, China) 2.66-22.33 (Jiang et al., 2011) Asia Huangpu River (Shanghai, China) 4.9-112.5 0.07-0.71 (Chen and Zhou, 2014) Asia Pearl River (China) 18.0* n.d.* (Liang et al., 2013) Asia Laizhou Bay-river discharge points (China) n.d.-18.7 (Zhang et al., 2012) Asia Brahmaputra River (Bangladesh) < 0.01-0.58 (Hossain et al., 2018) Asia Luan River (North China) 0.35-10.86 (Li et al., 2014) Europe Ebro River (Northeast Spain) 0.7-6.4 (García-Galán et al., 2011) Europe Lis River (Leiria, Portugal) n.d. (Paíga et al., 2016) Sulfamethoxazole Asia Pearl River Estuary (Zhuhai City, China) n.d.-195 0.37–219 (Li et al., 2018) Asia Yellow River (China) n.d. (Zhou et al., 2011) Asia Hai River (China) n.d. (Zhou et al., 2011) Asia Liao River (China) n.d. - <LOQ (Zhou et al., 2011) Asia Huangpu River (Shangai, China) 6.78-28.34 (Jiang et al., 2011) Asia Huangpu River (Shanghai, China) 2.2-764.9 0.05-0.6 (Chen and Zhou, 2014) Asia Laizhou Bay-river discharge points (China) 0.36 - 527 (Zhang et al., 2012) Asia Pearl River (China) 37.6* n.d. (Liang et al., 2013) Asia Chao Phraya River (Thailand) 2* (Tewari et al., 2013) Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 106 Meyer, B., Pailler, J.Y., Guignard, C., Hoffmann, L., Krein, A., 2011. Concentrations of dissolved herbicides and pharmaceuticals in a small river in Luxembourg. Environ. Monit. Assess. 180, 127–146. https://doi.org/10.1007/s10661-010-1777-9 Paíga, P., Santos, L.H.M.L.M., Delerue-Matos, C., 2017. Development of a multi-residue method for the determination of human and veterinary pharmaceuticals and some of their metabolites in aqueous environmental matrices by SPE-UHPLC–MS/MS. J. Pharm. Biomed. Anal. 135, 75–86. https://doi.org/10.1016/j.jpba.2016.12.013 Paíga, P., Santos, L.H.M.L.M., Ramos, S., Jorge, S., Silva, J.G., Delerue-Matos, C., 2016. Presence of pharmaceuticals in the Lis river (Portugal): Sources, fate and seasonal variation. Sci. Total Environ. 573, 164–177. https://doi.org/10.1016/j.scitotenv.2016.08.089 Pereira, A.M.P.T., Silva, L.J.G., Laranjeiro, C.S.M., Meisel, L.M., Lino, C.M., Pena, A., 2017. Human pharmaceuticals in Portuguese rivers: The impact of water scarcity in the environmental risk. Sci. Total Environ. 609, 1182–1191. https://doi.org/10.1016/j.scitotenv.2017.07.200 Ribeiro, C., Couto, C., Ribeiro, A.R., Maia, A.S., Santos, M., Tiritan, M.E., Pinto, E., Almeida, A.A., 2018. Distribution and environmental assessment of trace elements contamination of water, sediments and flora from Douro River estuary, Portugal. Sci. Total Environ. 639, 1381–1393. https://doi.org/10.1016/j.scitotenv.2018.05.234 Rocha, M.J., Ribeiro, M., Ribeiro, C., Couto, C., Cruzeiro, C., Rocha, E., 2012. Endocrine disruptors in the Leça River and nearby Porto Coast (NW Portugal): Presence of estrogenic compounds and hypoxic conditions. Toxicol. Environ. Chem. 94, 262–274. https://doi.org/10.1080/02772248.2011.644291 Santos, L.H.M.L.M., Ramalhosa, M.J., Ferreira, M., Delerue-Matos, C., 2016. Development of a modified acetonitrile-based extraction procedure followed by ultra-high performance liquid chromatography-tandem mass spectrometry for the analysis of psychiatric drugs in sediments. J. Chromatogr. A 1437, 37–48. https://doi.org/10.1016/j.chroma.2016.01.079 Tamtam, F., Mercier, F., Le Bot, B., Eurin, J., Tuc Dinh, Q., Clément, M., Chevreuil, M., 2008. Occurrence and fate of antibiotics in the Seine River in various hydrological conditions. Sci. Total Environ. 393, 84–95. https://doi.org/10.1016/j.scitotenv.2007.12.009 Ternes, T.A., 1998. Occurrence of drugs in German sewage treatment plants and rivers. Water Res. 32, 3245–3260. https://doi.org/10.1016/S0043-1354(98)00099-2 Tewari, S., Jindal, R., Kho, Y.L., Eo, S., Choi, K., 2013. Major pharmaceutical residues in wastewater treatment plants and receiving waters in Bangkok, Thailand, and associated ecological risks. Chemosphere 91, 697–704. https://doi.org/10.1016/j.chemosphere.2012.12.042 Wu, M., Xiang, J., Que, C., Chen, F., Xu, G., 2015. Occurrence and fate of psychiatric pharmaceuticals in the urban water system of Shanghai, China. Chemosphere 138, 486–493. https://doi.org/10.1016/j.chemosphere.2015.07.002 Zhang, R., Zhang, G., Zheng, Q., Tang, J., Chen, Y., Xu, W., Zou, Y., Chen, X., 2012. Occurrence and risks of antibiotics in the Laizhou Bay, China: Impacts of river discharge. Ecotoxicol. Environ. Saf. 80, 208–215. https://doi.org/10.1016/j.ecoenv.2012.03.002 Zhao, J.L., Ying, G.G., Liu, Y.S., Chen, F., Yang, J.F., Wang, L., Yang, X.B., Stauber, J.L., Warne, M.S.J., 2010. Occurrence and a screening-level risk assessment of human pharmaceuticals in the Chapter 3 107 pearl river system, South China. Environ. Toxicol. Chem. 29, 1377–1384. https://doi.org/10.1002/etc.161 Zhou, L.J., Ying, G.G., Zhao, J.L., Yang, J.F., Wang, L., Yang, B., Liu, S., 2011. Trends in the occurrence of human and veterinary antibiotics in the sediments of the Yellow River, Hai River and Liao River in northern China. Environ. Pollut. 159, 1877–1885. https://doi.org/10.1016/j.envpol.2011.03.034 Chapter 3 108 Figure SM1. Schematic SPE and QuEChERS methodologies for the extraction of the pharmaceuticals under study in river water and sediment samples. SPE extraction (Paíga et al., 2017) QuEChERS methodology (Santos et al., 2016) Chapter 3 109 a) Antibiotics b) Psychiatric drugs Note: The overlap of the twenty-seven pharmaceuticals would be barely perceptible in the chromatogram, so the pharmaceuticals were divided into two chromatograms separating them by the therapeutic class. Figure SM2. Overlay chromatogram for a) antibiotics and b) psychiatric drugs in a standard mixture with a concentration of 500 µg/L in each pharmaceutical. 0,0 0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 5,5 6,0 6,5 min 0 25000 50000 75000 100000 125000 150000 175000 200000 225000 250000 275000 300000 325000 350000 375000 400000 17:Erythromycin 734,50>158,00(+) CE: -35,0 16:Azithromycin 749,30>83,20(+) CE: -55,0 15:Clarithromycin 748,20>158,10(+) CE: -33,0 14:Sulfamethoxazole 254,00>156,00(+) CE: -17,0 13:Sulfadiazine 250,90>156,00(+) CE: -18,0 11:Sulfamethazine 278,80>186,05(+) CE: -20,0 10:sulfapyridine 249,80>92,05(+) CE: -30,0 9:Sulfamethoxypyridazine 280,80>156,00(+) CE: -19,0 8:Trimethoprim 290,80>230,10(+) CE: -26,0 7:Prulifloxacin 461,90>443,95(+) CE: -23,0 6:Lomefloxacin 352,00>334,10(+) CE: -22,0 5:Moxifloxacin 401,95>384,10(+) CE: -24,0 4:Norfloxacin 320,00>302,05(+) CE: -23,0 3:Ofloxacin 362,00>318,15(+) CE: -20,0 2:Enrofloxacin 359,90>342,15(+) CE: -23,0 1:Ciprofloxacin 331,90>314,10(+) CE: -23,0 0,0 0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 5,5 6,0 6,5 min 0 250000 500000 750000 1000000 1250000 1500000 1750000 2000000 2250000 28:10,11-epoxi carbamazepine 253,00>180,05(+) CE: -27,0 27:Diazepam 284,95>154,10(+) CE: -29,0 26:Trazodone 371,95>176,00(+) CE: -25,0 25:Paroxetine 330,00>44,05(+) CE: -28,0 24:Venlafaxine 278,00>58,00(+) CE: -20,0 23:citalopram 324,95>109,10(+) CE: -30,0 22:Sertraline 305,95>274,90(+) CE: -15,0 21:Fluoxetine 310,05>44,00(+) CE: -15,0 20:Carbamazepine 236,95>194,10(+) CE: -20,0 19:norfluoxetine 296,00>134,10(+) CE: -9,0 Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal 110 Figure SM3. The risk quotients (RQ) for different trophic levels (algae, Daphnia magna, and fish) of antibiotics and psychiatric drugs in the surface waters. Bold line marks risk quotient equal to one. Chapter 4 111 CHAPTER 4 GLYPHOSATE, AMPA AND GLUFOSINATE OCCURRENCE IN WATERS AND SEDIMENTS – WORLD ASSESSMENT As previously mentioned in Chapter 2, glyphosate has been the subject of legal controversy in recent years concerning its safety and potential link to cancer (Novotny, 2022). In 2015, the International Agency for Research on Cancer (IARC), of the World Health Organization (WHO), classified glyphosate as "probably carcinogenic to humans" based on studies that showed an association between glyphosate exposure and an increased risk of non-Hodgkin's lymphoma. This classification led to a series of lawsuits against the manufacturers of glyphosate-based herbicides, most notably Monsanto (now owned by Bayer). However, the US Environmental Protection Agency (EPA) and other regulatory agencies have generally affirmed that glyphosate is safe when used according to label instructions and have not restricted its use. Despite the possibility of posing a risk to the environment and to human and animals health, the European Commission (EC) recently renewed the use of glyphosate-based herbicides until December 15 of, 2023, approving a oneyear extension on the previous five-year authorization (European Commission, 2017). On the other hand, the use of glufosinate is already banned in Europe since 2020 (European Commission, 2020a, 2022a). Considering the alleged reprotoxicity, among other tested risks to human health related to the exposure to glufosinate, in 2017, the French Health and Environment Regulator (ANSES) had already withdrawn the selling authorization for this product in the French market, that included all the plant protection products containing glufosinate (ANSES, 2017). After their application to the soil/plant, they tend to degrade rapidly into their degradation products. Glyphosate is degraded, mostly by microbial action, taking the form of sarcosine acid and aminomethylphosphonic acid (AMPA), the last being its main metabolite and highly persistent in the environment (Stephen O. Duke, 2020). The first published study on glyphosate monitoring in groundwater dates back to 1996 (Smith et al., 1996) and in surface water to 1998 (Skark et al., 1998), this last reporting also AMPA’s contamination. Since then, concern about contamination by these compounds has been growing over the years, coupled with an increase of environmental sampling studies. To assess and evaluate glyphosate, AMPA, and glufosinate world’s environmental contamination, the published studies between January 1, 2018, and March 30, 2023, including glyphosate, AMPA, and glufosinate monitoring in water (surface and groundwater) and Glyphosate, AMPA and Glufosinate occurrence in waters and sediments – world assessment 112 sediments were analyzed. An extensive search on academic databases, such as Scopus, Web of Science, Science Direct, PubMed, Research Gate, and Google Scholar, was conducted using the combination of the following keywords: “glyphosate”, “glufosinate”, “aminomethylphosphonic acid”, or “AMPA” with “river”, "water resource", “riverside”, "hydrographic basin", “watercourse”, “freshwater”, “river basin”, “stream”, “tributary”, “lake”, “pond”, “groundwater”, “surface water”, “creeks”, “wetland”. After removing duplicates, the publications screening was performed by examining the entire content of the articles and gathering the relevant information (e.g., concentrations, analytical methods, number of samples, detection frequency) of the three compounds under study. Articles presenting model-based predictions of contaminations without information on the measured occurrences of compounds were excluded. Ninety-four publications were selected through more than 1 000 analyzed articles; the information contained therein is reviewed below. 4.1 Limits and threshold for Glyphosate, AMPA, and Glufosinate in water In order to analyze the concentrations of glyphosate, AMPA, and glufosinate found in waters and sediments around the world, a summary of the relevant legislation is of utmost importance. The applicable legislation to contaminations associated to these pollutants differs widely from country to country. Additionally, the same country may have different limit values depending on the type of water (drinking water, surface water, or groundwater). The restrictions can be targeted to individual and specified compounds, any pesticide individually, or even the sum of all pesticides found. The Figure 4.1 shows the maximum glyphosate concentration values allowed in water intended for human consumption established for some countries by current legislation. In Europe, the Council Directive 98/83/EC on the quality of water intended for human consumption provides a limit of 0.1 𝜇g L-1 (maximum individual value) to groundwater contamination by “pesticides and their relevant metabolites, degradation and reaction products” and of 0.5 𝜇g L-1 to the sum of all individual pesticides. Additionally, the directive states that “only those pesticides which are likely to be present in a given supply need be monitored“, leaving it unclear which compounds need to be monitored (European Commission, 1998). The European Union (EU) countries apply this directive and its amendments. Additionally, some EU countries have adapted their legislation based on European directives, as is the case of France, where in addition to the 0.1 𝜇g L-1 defined for Chapter 4 113 groundwater, it also defines environmental quality standards (EQS) to be taken into account in the assessment of the ecological status of continental surface waters (not intended for drinking water production) of 28 𝜇g L-1 for glyphosate and 452 𝜇g L-1 for AMPA. Also, propose a maximum acceptable value of 70 𝜇g L-1 for glyphosate and 45200 𝜇g L-1 for AMPA. For marine water establish an environmental guide value of 5.6 𝜇g L-1 and 45 𝜇g L-1 for glyphosate and AMPA, respectively with a maximum acceptable concentration value of 14 𝜇g L-1 and 4520 𝜇g L-1, respectively (Ministre de l’écologie du développement durable et de l’énergie, 2015; Portail Substaces Chimiques, 2020). Figure 4.1 – Legal limits for glyphosate residues in water intended for human consumption (developed by the author). In the USA, Environmental Protection Agency (EPA) national primary drinking water regulations also considered glyphosate in the list of organic contaminants in drinking water with a maximum contamination level of 700 𝜇g L-1 (EPA, 2009). Canada guidelines fixed the maximum acceptable concentration of glyphosate in drinking water of 280 𝜇g L-1 (Health Canada, 2022). Argentina established similar drinking water quality guideline levels for glyphosate, namely 300 𝜇g L-1 and a quality guide level for aquatic biota protection of 240 𝜇g L-1, both expressed as glyphosate isopropylamine (Subsecretaria de Recursos Hídricos de la Nación, 2003). Regarding Australian regulation, recommended maximum level for glyphosate presence in water is 1000 𝜇g/L. In this case, the legislated values are not generalized to all pesticides Glyphosate, AMPA and Glufosinate occurrence in waters and sediments – world assessment 114 but are individualized considering their characteristics and potential presence in the environment (NHMRC & NRMMC, 2011). In China, water bodies are divided into five classes according to the utilization purposes and protection objectives: class I refers to spring water and national nature reserves; class II and III are the first and second classes of protected areas for centralized sources of drinking water, for aquatic species and swimming areas; class IV refers to water areas for industrial and non-direct exposure recreation uses; and class V is mainly applicable to the water bodies for agricultural use and landscape (Asian Development Bank, 2016). Also, groundwater is similarly classified in the Standard for groundwater quality (General Administration of Quality Supervision, 2017). Environmental Quality Standards for Surface Water apply to the different classes of surface water. GB 5749-2006, the standards for drinking Water Quality concerning drinking water sources (class II and III), presents a limit especially to glyphosate, of 700 𝜇g L-1 (Ministry of Health of China, 2006). Japan legislation exhibits one of the highest limit values for glyphosate concentration in water, of 2 000 𝜇g L-1. This legal framework is the particularity that the indicated limit refers to the sum of glyphosate and AMPA’s concentration. Additionally, in 2014 a review of the legislation included the addition of a maximum concentration value for glufosinate contamination of 20 𝜇g L-1 (Ministry of Health Labor and Welfare of Japan, 2003). In Brazil, there are two distinct limits for the contamination of water intended for human consumption. The first is presented in the applicable legislation for surface waters which, similarly to China, presents a five-class categorization taking into account the surface water destination: special class, water intended for drinking water supply, with disinfection as well as the preservation of the natural balance of aquatic communities and environments; class 1, intended for drinking water supply, after simplified treatment, as well as the protection of aquatic communities, recreational activities of primary contact (e.g. swimming, diving) and irrigation of raw and direct consumption vegetables; class 2, destined for the drinking water supply, after conventional treatment as well as the protection of aquatic communities, recreational activities of primary contact and the irrigation of vegetables, fruit plants among others with which the public may have direct contact; class 3, destined for the drinking water supply, after conventional or advanced treatment, irrigation and recreational activities of secondary contact; and class 4, destined for navigation; and for landscape harmony. For classes 1 and 2, the limit value for glyphosate concentration is 65 𝜇g L-1, and for class 3, 280 𝜇g L-1 (CONAMA, 2005). For water intended for human consumption supplied from an alternative water supply system or solution, it is established a different limit of 500 𝜇g L-1 not Chapter 4 115 only for glyphosate concentration in water but for the sum of glyphosate and AMPA’s concentration (Ministério da Saúde, 2017). Given the evident differences found in the maximum limits for glyphosate and AMPA in water intended for human consumption and, in some cases, the absence of any one or all three in their legal documents associated with water quality, several authors refer to the importance and urgency of revise the current legal limits of glyphosate, AMPA and glufosinate in surface and groundwaters for human and aquatic biota protection (Campanale et al., 2021; D. M. de Oliveira et al., 2023; Z. Li, 2018; Z. Li & Fantke, 2022). 4.2 Pre-treatment, extraction, and analysis Accurate measurement of glyphosate, AMPA, and glufosinate concentrations in environmental samples is essential for quantitative exposure assessments. However, the unique chemical properties of these compounds, such as high-water solubility, low solubility in organic solvents and volatilization, absence of chromophores or fluorophores, ionic character and their preference for forming complexes turns extraction and quantification into a challenging process (EFSA, 2005, 2015; Valle et al., 2019). Reviewing the analytical methods applied in the environmental monitoring of waters and sediments contamination by glyphosate, AMPA, and glufosinate published between January 2018 and March 2023 worldwide, it can be seen that the most applied detection methods can be divided into three main categories: high-performance, ultra-performance or Ultra-High- Performance liquid chromatography (HPLC, UPLC or UHPLC) (Alonso et al., 2022; Lajmanovich et al., 2023; López-Vázquez et al., 2023; Raby et al., 2022; Rodríguez-Bolaña et al., 2023; Suciu et al., 2023; Natalia Veronica Van Opstal et al., 2023), gas-chromatography (GC) (Donald et al., 2018; Rezaei Kalantary et al., 2022; Tongo et al., 2022), and enzyme-linked immunosorbent assay (ELISA) (Horn et al., 2019; Melendez-Pastor et al., 2021; Reynoso et al., 2020; Welch et al., 2019). Figure 4.2 resumes all analytical methods used in the studies and shows that highperformance (HPLC – 38%) and ultra-performance or Ultra-High-Performance (UHPLC/UPLC- 36%) liquid chromatography were the most applied methodologies, followed by LC (7%), ELISA (4%) and gas chromatography (GC-3%). References 218 consumption. Official Journal of the European Union, L 260, 6–17. European Commission. (2015b). Commission Implementing Decision (EU) 2015/495 of 20 March 2015 establishing a watch list of substances for Union-wide monitoring in the field of water policy pursuant to Directive 2008/105/EC of the European Parliament and of t. Official Journal of European Union L 78, L 78, 40–42. https://eur-lex.europa.eu/legalcontent/EN/TXT/?uri=uriserv%3AOJ.L_.2015.078.01.0040.01.ENG European Commission. (2017). Commission implementing regulation (EU) 2017/2324 of 12 December 2017 renewing the approval of the active substance glyphosate in accordance with Regulation (EC) No 1107/2009 of the European Parliament and of the Council concerning the placing of plant pr. Official Journal of the European Union, L 333, 10–16. https://webarchive.nationalarchives.gov.uk/eu-exit/20200124174550/https://eurlex.europa.eu/legal-content/EN/TXT/?uri=CELEX:32017R2324 European Commission. (2018). Commission Implementing Decision (EU) 2018/840 of 5 June 2018 establishing a watch list of substances for Union-wide monitoring in the field of water policy pursuant to Directive 2008/105/EC of the European Parliament. Official Journal of the European Union, L 141, 9–12. https://eur-lex.europa.eu/legalcontent/EN/TXT/?uri=CELEX:32018D0840 European Commission. (2020a). Commission delegated Regulation 2020/1068 amending Annexes I and V to Regulation (EU) No 649/2012 of the European Parliament and of the Council concerning the export and import of hazardous chemicals. Official Journal of the European Union, L 234, 1–7. https://ec.europa.eu/food/plant/pesticides/eu-pesticides- database/start/screen/active-substances/details/79 European Commission. (2020b). Commission Implementing Decision (Eu) 2020/1161 of 4 August 2020 establishing a watch list of substances for Union-wide monitoring in the field of water policy pursuant to Directive 2008/105/EC of the European Parliament and of the Council. Official Journal of the European Union, L 257, 32–35. https://eurlex.europa.eu/legalcontent/EN/TXT/?uri=uriserv:OJ.L_.2020.257.01.0032.01.ENG&toc=OJ:L:2020:257:TOC European Commission. (2022a). Commission Implementing Regulation (EU) 2022/801 of 20 May 2022 amending Implementing Regulation (EU) No 540/2011 to update the list of active substances approved or deemed to have been approved under Regulation (EC) No 1107/2009 of the European Parliamen. Official Journal of the European Union, L143, 7– 10. https://eur-lex.europa.eu/eli/reg_impl/2022/801/oj European Commission. (2022b). Glyphosate. https://food.ec.europa.eu/plants/pesticides/approval-active-substances/renewalapproval/glyphosate_en European Commission. (2023). Glyphosate. https://food.ec.europa.eu/plants/pesticides/approval-active-substances/renewalapproval/glyphosate_en#status-of-glyphosate-in-the-eu European Environment Agency. (2019). The European environment - state and outlook 2020. Knowledge for transition to a sustainable Europe. In European Union (Vol. 60, Issue 6). European Environment Agency. https://doi.org/10.15196/TS600305 Maria João Cerveira Fernandes 219 European Standard. (2011). EN15290:2011 – Solid biofuels – Determination of major elements – Al, Ca, Fe, Mg, P, K, Si, Na and Ti. European Committee for Standardization. Eurostat. (2022). Pesticide sales. Eurostat. https://ec.europa.eu/eurostat/databrowser/view/aei_fm_salpest09/default/table?lang =e Evagelista, P. A., Vilca, F. Z., Pimpinato, R. F., & Tornisielo, V. L. (2020). Optimization of a methodology for simultaneous determination of four antidepressants present in fresh water by high efficiency liquid chromatography. Revista Eletrônica Em Gestão, Educação e Tecnologia Ambiental, 24, e8. https://doi.org/10.5902/2236117062663 Fabbri, D., Pizzol, R., Calza, P., Malandrino, M., Gaggero, E., Padoan, E., & Ajmone-Marsan, F. (2021). Constructed technosols: A strategy toward a circular economy. Applied Sciences (Switzerland), 11(8). https://doi.org/10.3390/app11083432 FAO. (2022). FAOSTAT: Pesticides Use. License: CC BY-NC-SA 3.0 IGO. https://www.fao.org/faostat/en/#data/RP Federal Office of Consumer Protection and Food Safety. (2021). Sales quantities of active substances in plant protection products from 1987 to 2021 (in German). https://www.bvl.bund.de/EN/Tasks/04_Plant_protection_products/01_ppp_tasks/02_p pp_AuthorisationReviewActSub/03_ppp_DomesticSalesExport/PPP_domesticSales_and _Export_node.html Fehsenfeld, T. (2015). The Effect of Soil Type on Migration of Pharmaceuticals and Personal Care Products (PPCPs) Through Cape Cod Soils. https://www.mbl.edu/ses/files/2016/02/Theo_ProjectPaperFinalforweb.pdf Feng, D., Soric, A., & Boutin, O. (2020). Treatment technologies and degradation pathways of glyphosate: A critical review. Science of the Total Environment, 742, 140559. https://doi.org/10.1016/j.scitotenv.2020.140559 Fernandes, G., Aparicio, V. C., Bastos, M. C., De Gerónimo, E., Labanowski, J., Prestes, O. D., Zanella, R., & dos Santos, D. R. (2019). Indiscriminate use of glyphosate impregnates river epilithic biofilms in southern Brazil. Science of the Total Environment, 651, 1377–1387. https://doi.org/10.1016/j.scitotenv.2018.09.292 Fernandes, M. J., Moreira, M. M., Paíga, P., Dias, D., Bernardo, M., Carvalho, M., Lapa, N., Fonseca, I., Morais, S., Figueiredo, S., & Delerue-Matos, C. (2019). Evaluation of the adsorption potential of biochars prepared from forest and agri-food wastes for the removal of fluoxetine. Bioresource Technology, 292(August), 121973. https://doi.org/10.1016/j.biortech.2019.121973 Ferreiro, D. A. (2017). Recuperación de suelos contaminados con aniones potencialmente tóxicos y metales pesados mediante Tecnosoles y biocarbones adsorbentes [Universidade de Santiago de Compostela]. http://hdl.handle.net/10347/16427 Fogliatto, S., Ferrero, A., & Vidotto, F. (2020). Current and future scenarios of glyphosate use in Europe: Are there alternatives? In D. L. Sparks (Ed.), Advances in Agronomy (1st ed., Vol. 163, pp. 219–278). Elsevier Inc. https://doi.org/10.1016/bs.agron.2020.05.005 References 220 Freundlich, H. (1907). Über die Adsorption in Lösungen. Zeitschrift Für Physikalische Chemie, 57U, 385. https://doi.org/doi:10.1515/zpch-1907-5723 Fryda, L., & Visser, R. (2015). Biochar for Soil Improvement: Evaluation of Biochar from Gasification and Slow Pyrolysis. Agriculture (Switzerland), 5(4), 1076–1115. https://doi.org/10.3390/agriculture5041076 Garba, J., Saiful, M., & Hamdani, A. (2018). Simplified method for derivatization of extractable glyphosate and aminomethylphosphonic acid and their determination by high performance liquid chromatography Environmental Research & Technology. Environmental Research & Technology, 1(2), 19–30. Garba, J., Samsuri, A. W., Othman, R., & Ahmad Hamdani, M. S. (2018). Adsorption-desorption and leaching potential of glyphosate and aminomethylphosphonic acid in acidic Malaysian soil amended with cow dung and rice husk ash. Environmental Monitoring and Assessment, 190(11). https://doi.org/10.1007/s10661-018-7034-3 Gasco Cavero, S., García-Gil, A., Cruz-Pérez, N., Martín Rodríguez, L. F., Laspidou, C., Contreras- Llin, A., Quintana, G., Díaz-Cruz, S., & Santamarta, J. C. (2023). First emerging pollutants profile in groundwater of the volcanic active island of El Hierro (Canary Islands). Science of the Total Environment, 872(October 2022). https://doi.org/10.1016/j.scitotenv.2023.162204 Geerdink, R. B., Hassing, M., Ayarza, N., Bruggink, C., Wielheesen, M., Claassen, J., & Epema, O. J. (2020). Analysis of glyphosate, AMPA, Glufosinate and MPPA with ION chromatography tandem mass spectrometry using A membrane suppressor in the ammonium form application to surface water of low to moderate salinity. Analytica Chimica Acta, 1133, 66–76. https://doi.org/10.1016/j.aca.2020.05.058 General Administration of Quality Supervision, I. and Q. of the P. (2017). Standard for groundwater quality (GB/T 14848-2017). National Standard of the People’s Republic of China, 1–22. https://www.chinesestandard.net/PDF.aspx/GBT14848-2017 Geng, Y., Jiang, L., Zhang, D., Liu, B., Zhang, J., Cheng, H., Wang, L., Peng, Y., Wang, Y., Zhao, Y., Xu, Y., & Liu, X. (2021). Glyphosate, aminomethylphosphonic acid, and glufosinate ammonium in agricultural groundwater and surface water in China from 2017 to 2018: Occurrence, main drivers, and environmental risk assessment. Science of the Total Environment, 769, 144396. https://doi.org/10.1016/j.scitotenv.2020.144396 Gerónimo, E. De, Aparicio, V. C., & Costa, J. L. (2018). Glyphosate sorption to soils of Argentina. Estimation of affinity coefficient by pedotransfer function. Geoderma, 322, 140–148. https://doi.org/10.1016/j.geoderma.2018.02.037 Ghavamifar, S., Naidu, R., Mozafari, V., & Li, Z. (2023). Can calcite play a role in the adsorption of glyphosate? A comparative study with a new challenge. Chemosphere, 311(P1), 136922. https://doi.org/10.1016/j.chemosphere.2022.136922 Giles, C. H., Smith, D., & Huitson, A. (1974). A General Treatment and Classification of the Solute Adsorption Isotherm. Journal of Colloid and Interface Science, 47(3), 755–765. https://doi.org/https://doi.org/10.1016/0021-9797(74)90252-5 Gillezeau, C., Van Gerwen, M., Shaffer, R. M., Rana, I., Zhang, L., Sheppard, L., & Taioli, E. Maria João Cerveira Fernandes 221 (2019). The evidence of human exposure to glyphosate: A review. Environmental Health: A Global Access Science Source, 18(1), 1–14. https://doi.org/10.1186/s12940-018-0435- 5 Gimsing, A. L., Borggaard, O. K., & Bang, M. (2004). Influence of soil composition on adsorption of glyphosate and phosphate by contrasting Danish surface soils. European Journal of Soil Science, 55(1), 183–191. https://doi.org/10.1046/j.1365-2389.2003.00585.x Glinski, D. A., Purucker, S. T., Van Meter, R. J., Black, M. C., & Henderson, W. M. (2018). Analysis of pesticides in surface water, stemflow, and throughfall in an agricultural area in South Georgia, USA. Chemosphere, 209, 496–507. https://doi.org/10.1016/j.chemosphere.2018.06.116 Goldan, E., Nedeff, V., Barsan, N., Culea, M., Tomozei, C., Panainte-Lehadus, M., & Mosnegutu, E. (2022). Evaluation of the Use of Sewage Sludge Biochar as a Soil Amendment—A Review. Sustainability (Switzerland), 14(9). https://doi.org/10.3390/su14095309 Gomes, A. I., Pires, J. C. M., Figueiredo, S. A., & Boaventura, R. A. R. (2014). Optimization of River Water Quality Surveys by Multivariate Analysis of Physicochemical, Bacteriological and Ecotoxicological Data. Water Resources Management, 28(5), 1345–1361. https://doi.org/10.1007/s11269-014-0547-9 Gomes, M. P., Brito, J. C. M., Vieira, F., Kitamura, R. S. A., & Juneau, P. (2022). Emerging Contaminants in Streams of Doce River Watershed, Minas Gerais, Brazil. Frontiers in Environmental Science, 9(January), 1–11. https://doi.org/10.3389/fenvs.2021.801599 Grandcoin, A., Piel, S., & Baurès, E. (2017). AminoMethylPhosphonic acid (AMPA) in natural waters: Its sources, behavior and environmental fate. Water Research, 117, 187–197. https://doi.org/10.1016/j.watres.2017.03.055 Grau, D., Grau, N., Gascuel, Q., Paroissin, C., Stratonovitch, C., Lairon, D., Devault, D. A., & Di Cristofaro, J. (2022). Quantifiable urine glyphosate levels detected in 99% of the French population, with higher values in men, in younger people, and in farmers. Environmental Science and Pollution Research, 29(22), 32882–32893. https://doi.org/10.1007/s11356- 021-18110-0 Grilli, E., Vigliotti, R. C., Fiorentino, A., Scognamiglio, M., Rossetti, L., Nogueira, T. A. R., Jani, A. D., Abreu-Junior, C. H., Ribeiro Roder, L., Ganga, A., & Capra, G. F. (2023). Constructed Technosols as a Soil Rebuilding Technique to Reclaim Abandoned Limestone Quarries in the Mediterranean Region: A Field Study. Sustainability, 15(6), 5036. https://doi.org/10.3390/su15065036 Gros, P., Ahmed, A., Kühn, O., & Leinweber, P. (2017). Glyphosate binding in soil as revealed by sorption experiments and quantum-chemical modeling. Science of the Total Environment, 586, 527–535. https://doi.org/10.1016/j.scitotenv.2017.02.007 Gunarathna, S., Gunawardana, B., Jayaweera, M., Manatunge, J., & Zoysa, K. (2018). Glyphosate and AMPA of agricultural soil, surface water, groundwater and sediments in areas prevalent with chronic kidney disease of unknown etiology, Sri Lanka. Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural Wastes, 53(11), 729–737. https://doi.org/10.1080/03601234.2018.1480157 References 222 Guo, M., Song, W., & Tian, J. (2020). Biochar-Facilitated Soil Remediation: Mechanisms and Efficacy Variations. Frontiers in Environmental Science, 8(October). https://doi.org/10.3389/fenvs.2020.521512 Hall, K. E., Spokas, K. A., Gamiz, B., Cox, L., Papiernik, S. K., & Koskinen, W. C. (2018). Glyphosate sorption/desorption on biochars – interactions of physical and chemical processes. Pest Management Science, 74(5), 1206–1212. https://doi.org/10.1002/ps.4530 Hall, K. R., Eagleton, L. C., Acrivos, A., & Vermeulen, T. (1966). Pore- and solid-diffusion kinetics in fixed-bed adsorption under constant-pattern conditions. Industrial and Engineering Chemistry Fundamentals, 5(2), 212–223. https://doi.org/10.1021/i160018a011 Hanke, I., Singer, H., & Hollender, J. (2008). Ultratrace-level determination of glyphosate, aminomethylphosphonic acid and glufosinate in natural waters by solid-phase extraction followed by liquid chromatography-tandem mass spectrometry: Performance tuning of derivatization, enrichment and detection. Analytical and Bioanalytical Chemistry, 391(6), 2265–2276. https://doi.org/10.1007/s00216-008-2134-5 Hawash, H. B., Moneer, A. A., Galhoum, A. A., Elgarahy, A. M., Mohamed, W. A. A., Samy, M., El-Seedi, H. R. M. S. G., Mubarak, M. F., & F.Attia, N. (2023). Occurrence and spatial distribution of pharmaceuticals and personal care products (PPCPs) in the aquatic environment, their characteristics, and adopted legislations. Journal of Water Process Engineering, 52(December 2022), 103490. https://doi.org/10.1016/j.jwpe.2023.103490 Health Canada. (2022). Guidelines for Canadian Drinking Water Quality—Summary Tables. Healthy Environments and Consumer Safety Branch, 1–42. https://doi.org/10.1021/ba- 1987-0214.ch035 Hedde, M., Nahmani, J., Séré, G., Auclerc, A., & Cortet, J. (2019). Early colonization of constructed Technosols by macro-invertebrates. Journal of Soils and Sediments, 19(8), 3193–3203. https://doi.org/10.1007/s11368-018-2142-9 Herath, I., Kumarathilaka, P., Al-Wabel, M. I., Abduljabbar, A., Ahmad, M., Usman, A. R. A., & Vithanage, M. (2016). Mechanistic modeling of glyphosate interaction with rice husk derived engineered biochar. Microporous and Mesoporous Materials, 225, 280–288. https://doi.org/10.1016/j.micromeso.2016.01.017 Ho, Y. S., & McKay, G. (1999). Pseudo-second order model for sorption processes. Process Biochemistry, 34(5), 451–465. https://doi.org/10.1016/S0032-9592(98)00112-5 Hoerlein, G. (1994). Glufosinate (phosphinothricin), a natural amino acid with unexpected herbicidal properties. Reviews of Environmental Contamination and Toxicology, 138, 73– 145. https://doi.org/10.1007/978-1-4612-2672-7_4 Horn, S., Pieters, R., & Bøhn, T. (2019). A first assessment of glyphosate, 2,4-D and Cry proteins in surface water of South Africa. South African Journal of Science, 115(9–10), 1–7. https://doi.org/10.17159/sajs.2019/5988 Huhn, C. (2018). More and enhanced glyphosate analysis is needed. Analytical and Bioanalytical Chemistry, 410(13), 3041–3045. https://doi.org/10.1007/s00216-018- 1000-3 Maria João Cerveira Fernandes 223 Huntscha, S., Stravs, M. A., Bühlmann, A., Ahrens, C. H., Frey, J. E., Pomati, F., Hollender, J., Buerge, I. J., Balmer, M. E., & Poiger, T. (2018). Seasonal Dynamics of Glyphosate and AMPA in Lake Greifensee: Rapid Microbial Degradation in the Epilimnion during Summer. Environmental Science and Technology, 52(8), 4641–4649. https://doi.org/10.1021/acs.est.8b00314 IARC. (2017). Some organophosphate insecticides and herbicides. In Monographs on the evaluation of carcinogenics risks to humans. (Vol. 112, p. 451). International Agency for Research on Cancer. http://monographs.iarc.fr/ENG/Publications/corrigenda.php IBAMA. (2022). Painel de Informações sobre a Comercialização de Agrotóxicos e Afins no Brasil – série histórica 2009 - 2021. Instituto Brasileiro Do Meio Ambiente e Dos Recursos Naturais Renováveis (IBAMA). https://www.gov.br/ibama/pt-br/assuntos/quimicos-e- biologicos/agrotoxicos/paineis-de-informacoes-de-agrotoxicos/paineis-de-informacoes- de-agrotoxicos#Painel-comercializacao Ighalo, J. O., Ajala, O. J., Adeniyi, A. G., Babatunde, E. O., & Ajala, M. A. (2021). Ecotoxicology of glyphosate and recent advances in its mitigation by adsorption. Environmental Science and Pollution Research, 28(3), 2655–2668. https://doi.org/10.1007/s11356-020-11521-5 Ishfaq Showket Mir, Monaza Rashid, Javeed Ahmad Khan, & Bisma Yousuf. (2022). Emerging Pollutants- A review of current understanding and future scenario. World Journal of Advanced Research and Reviews, 16(2), 074–088. https://doi.org/10.30574/wjarr.2022.16.2.1119 IUSS Working Group WRB. (2015). World reference base for soil resources 2014, update 2015 - International soil classification system for naming soils and creating legends for soil maps. In World Soil Resources Reports No. 106. https://doi.org/10.1017/S0014479706394902 Jaria, G., Calisto, V., Gil, M. V., Otero, M., & Esteves, V. I. (2015). Removal of fluoxetine from water by adsorbent materials produced from paper mill sludge. Journal of Colloid and Interface Science, 448, 32–40. https://doi.org/10.1016/j.jcis.2015.02.002 Jelić, A., Gros, M., Petrovic, M., Ginebreda, A., & Barceló, D. (2012). Occurrence and Elimination of Pharmaceuticals During Conventional Wastewater Treatment. Emerging and Priority Pollutants in Rivers. The Handbook of Environmental Chemistry, 19, 1–24. https://doi.org/10.1007/978-3-642-25722-3 Jia, G., Xu, J., Long, X., Ge, S., Chen, L., Hu, D., & Zhang, Y. (2019). Enantioselective Degradation and Chiral Stability of Glufosinate in Soil and Water Samples and Formation of 3- Methylphosphinicopropionic Acid and N-Acetyl-glufosinate Metabolites. Journal of Agricultural and Food Chemistry, 67(41), 11312–11321. https://doi.org/10.1021/acs.jafc.9b01028 K’oreje, K. O., Vergeynst, L., Ombaka, D., De Wispelaere, P., Okoth, M., Van Langenhove, H., & Demeestere, K. (2016). Occurrence patterns of pharmaceutical residues in wastewater, surface water and groundwater of Nairobi and Kisumu city, Kenya. Chemosphere, 149, 238–244. https://doi.org/10.1016/j.chemosphere.2016.01.095 Kairigo, P., Ngumba, E., Sundberg, L. R., Gachanja, A., & Tuhkanen, T. (2020). Occurrence of References 224 antibiotics and risk of antibiotic resistance evolution in selected Kenyan wastewaters, surface waters and sediments. Science of the Total Environment, 720, 137580. https://doi.org/10.1016/j.scitotenv.2020.137580 Karungamye, P., Rugaika, A., Mtei, K., & Machunda, R. (2022). The pharmaceutical disposal practices and environmental contamination: A review in East African countries. HydroResearch, 5, 99–107. https://doi.org/10.1016/j.hydres.2022.11.001 Khan, A. H., Aziz, H. A., Khan, N. A., Hasan, M. A., Ahmed, S., Farooqi, I. H., Dhingra, A., Vambol, V., Changani, F., Yousefi, M., Islam, S., Mozaffari, N., & Mahtab, M. S. (2022). Impact, disease outbreak and the eco-hazards associated with pharmaceutical residues: a Critical review. International Journal of Environmental Science and Technology, 19(1), 677–688. https://doi.org/10.1007/s13762-021-03158-9 Khan, Afzal Husain, Khan, N. A., Zubair, M., Azfar Shaida, M., Manzar, M. S., Abutaleb, A., Naushad, M., & Iqbal, J. (2022). Sustainable green nanoadsorbents for remediation of pharmaceuticals from water and wastewater: A critical review. Environmental Research, 204, 112243. https://doi.org/https://doi.org/10.1016/j.envres.2021.112243 Kocadal, K., Alkas, F. B., Battal, D., & Saygi, S. (2022). A review on advances and perspectives of glyphosate determination: challenges and opportunities. Archives of Environmental Protection, 48(3), 89–98. https://doi.org/10.24425/aep.2022.142693 Kock, A., Glanville, H. C., Law, A. C., Stanton, T., Carter, L. J., & Taylor, J. C. (2023). Emerging challenges of the impacts of pharmaceuticals on aquatic ecosystems: A diatom perspective. Science of The Total Environment, 878(February), 162939. https://doi.org/10.1016/j.scitotenv.2023.162939 Koskinen, W. C., Marek, L. J., & Hall, K. E. (2016). Analysis of glyphosate and aminomethylphosphonic acid in water, plant materials and soil. Pest Management Science, 72(3), 423–432. https://doi.org/10.1002/ps.4172 Kosma, C. I., Kapsi, M. G., Konstas, P. S. G., Trantopoulos, E. P., Boti, V. I., Konstantinou, I. K., & Albanis, T. A. (2020). Assessment of multiclass pharmaceutical active compounds (PhACs) in hospital WWTP influent and effluent samples by UHPLC-Orbitrap MS: Temporal variation, removals and environmental risk assessment. Environmental Research, 191(August), 110152. https://doi.org/10.1016/j.envres.2020.110152 Kotwani, A., Joshi, J., & Kaloni, D. (2021). Pharmaceutical effluent: a critical link in the interconnected ecosystem promoting antimicrobial resistance. Environmental Science and Pollution Research, 28(25), 32111–32124. https://doi.org/10.1007/s11356-021- 14178-w Kubotera, H., Yamaguchi, N., & Kusaba, T. (2020). New definition of the qualifiers for Dystric and Eutric should be noted for the classification of Andosols with WRB2014. Soil Science and Plant Nutrition, 66(4), 593–601. https://doi.org/10.1080/00380768.2020.1783699 Kudsk, P., & Mathiassen, S. K. (2020). Pesticide regulation in the European Union and the glyphosate controversy. Weed Science, 68(3), 214–222. https://doi.org/10.1017/wsc.2019.59 Kudzin, M. H., Zylla, R., Mrozinska, Z., & Urbaniak, P. (2019). 31 P NMR investigations on Maria João Cerveira Fernandes 225 roundup degradation by AOP procedures. Water (Switzerland), 11(2). https://doi.org/10.3390/w11020331 Kumari, K. G. I. D., Moldrup, P., Paradelo, M., Elsgaard, L., & de Jonge, L. W. (2016). Soil Properties Control Glyphosate Sorption in Soils Amended with Birch Wood Biochar. Water, Air, and Soil Pollution, 227(6). https://doi.org/10.1007/s11270-016-2867-2 Kurakalva, R. M., & Aradhi, K. K. (2022). Occurrence and distribution of HCHs and DDTs in surface water and groundwater from the Gajulamandyam region along the Swarnamukhi river basin, Andhra Pradesh, India. International Journal of Environmental Analytical Chemistry, 102(18), 6955–6969. https://doi.org/10.1080/03067319.2020.1818735 la Cecilia, D., & Maggi, F. (2020). Influential sources of uncertainty in glyphosate biochemical degradation in soil. Mathematics and Computers in Simulation, 175(xxxx), 121–139. https://doi.org/10.1016/j.matcom.2020.01.003 Laghari, M., Naidu, R., Xiao, B., Hu, Z., Mirjat, M. S., Hu, M., Kandhro, M. N., Chen, Z., Guo, D., Jogi, Q., Abudi, Z. N., & Fazal, S. (2016). Recent developments in biochar as an effective tool for agricultural soil management: a review. Journal of the Science of Food and Agriculture, 96(15), 4840–4849. https://doi.org/10.1002/jsfa.7753 Lajin, B., & Goessler, W. (2019). Direct speciation analysis of organophosphorus environmental pollutants in water by HPLC-ICPMS/MS. Talanta, 196(December 2018), 357–361. https://doi.org/10.1016/j.talanta.2018.12.075 Lajmanovich, R. C., Repetti, M. R., Cuzziol Boccioni, A. P., Michlig, M. P., Demonte, L., Attademo, A. M., & Peltzer, P. M. (2023). Cocktails of pesticide residues in Prochilodus lineatus fish of the Salado River (South America): First record of high concentrations of polar herbicides. Science of the Total Environment, 870(October 2022). https://doi.org/10.1016/j.scitotenv.2023.162019 Langmuir, I. (1918). The adsorption of gases on plane surfaces of glass, mica and platinum. Journal of the American Chemical Society, 40(9), 1361–1403. https://doi.org/10.1021/ja02242a004 Lapworth, D. J., Baran, N., Stuart, M. E., & Ward, R. S. (2012). Emerging organic contaminants in groundwater: A review of sources, fate and occurrence. Environmental Pollution, 163, 287–303. https://doi.org/10.1016/j.envpol.2011.12.034 Lehmann, J. (2007). Bio‐energy in the black. Frontiers in Ecology and Environment, 5(7), 381– 387. Lehmann, J., Rillig, M. C., Thies, J., Masiello, C. A., Hockaday, W. C., & Crowley, D. (2011). Biochar effects on soil biota - A review. Soil Biology and Biochemistry, 43(9), 1812–1836. https://doi.org/10.1016/j.soilbio.2011.04.022 Lehmann, J., & Stephen, J. (2009). Biochar for environmental management : science and technology. In J. Lehmann & S. Joseph (Eds.), Biochar for Environmental Management. Earthscan. Li, N., He, M., Lu, X., Yan, B., Duan, X., Chen, G., Wang, S., & Hou, L. (2022). Municipal solid waste derived biochars for wastewater treatment: Production, properties and References 226 applications. Resources, Conservation and Recycling, 177(135), 106003. https://doi.org/10.1016/j.resconrec.2021.106003 Li, S., Shi, W., Li, H., Xu, N., Zhang, R., Chen, X., Sun, W., Wen, D., He, S., Pan, J., He, Z., & Fan, Y. (2018). Antibiotics in water and sediments of rivers and coastal area of Zhuhai City, Pearl River estuary, south China. Science of the Total Environment, 636, 1009–1019. https://doi.org/10.1016/j.scitotenv.2018.04.358 Li, X., He, F., Wang, Z., & Xing, B. (2022). Roadmap of environmental health research on emerging contaminants: Inspiration from the studies on engineered nanomaterials. Eco- Environment & Health, 1(3), 181–197. https://doi.org/10.1016/j.eehl.2022.10.001 Li, Y., Wang, F., Lin, Q., Deng, P., Zhang, Y., & Hu, D. (2022). The fate and behavior of glufosinate-enantiomers and their metabolites in open-field soil and weeds. Arabian Journal of Chemistry, 15(11), 104191. https://doi.org/10.1016/j.arabjc.2022.104191 Li, Z. (2018). A health-based regulatory chain framework to evaluate international pesticide groundwater regulations integrating soil and drinking water standards. Environment International, 121(August), 1253–1278. https://doi.org/10.1016/j.envint.2018.10.047 Li, Z., & Fantke, P. (2022). Toward harmonizing global pesticide regulations for surface freshwaters in support of protecting human health. Journal of Environmental Management, 301(October 2021), 113909. https://doi.org/10.1016/j.jenvman.2021.113909 Liang, X., Chen, B., Nie, X., Shi, Z., Huang, X., & Li, X. (2013). The distribution and partitioning of common antibiotics in water and sediment of the Pearl River Estuary, South China. Chemosphere, 92(11), 1410–1416. https://doi.org/10.1016/j.chemosphere.2013.03.044 Liao, F., Yang, L., Li, Q., Li, Y. R., Yang, L. T., Anas, M., & Huang, D. L. (2018). Characteristics and inorganic N holding ability of biochar derived from the pyrolysis of agricultural and forestal residues in the southern China. Journal of Analytical and Applied Pyrolysis, 134(August), 544–551. https://doi.org/10.1016/j.jaap.2018.08.001 Liao, Y., Berthion, J. M., Colet, I., Merlo, M., Nougadère, A., & Hu, R. (2018). Validation and application of analytical method for glyphosate and glufosinate in foods by liquid chromatography-tandem mass spectrometry. Journal of Chromatography A, 1549, 31– 38. https://doi.org/10.1016/j.chroma.2018.03.036 Lima, I. B., Boëchat, I. G., Fernandes, M. D., Monteiro, J. A. F., Rivaroli, L., & Gücker, B. (2022). Glyphosate pollution of surface runoff, stream water, and drinking water resources in Southeast Brazil. Environmental Science and Pollution Research, 0123456789. https://doi.org/10.1007/s11356-022-24167-2 Lin, J. F., Chang, F. C., & Sheen, J. F. (2022). Determination of glyphosate, aminomethylphosphonic acid, and glufosinate in river water and sediments using microwave-assisted rapid derivatization and LC–MS/MS. Environmental Science and Pollution Research, 29(30), 46282–46292. https://doi.org/10.1007/s11356-022-19189-9 Liu, B., Peng, S., Liao, Y., & Wang, H. (2019). The characteristics and causes of increasingly severe saltwater intrusion in Pearl River Estuary. Estuarine, Coastal and Shelf Science, 220(November 2018), 54–63. https://doi.org/10.1016/j.ecss.2019.02.041 Maria João Cerveira Fernandes 227 Loganathan, P., Vigneswaran, S., Kandasamy, J., Cuprys, A. K., Maletskyi, Z., & Ratnaweera, H. (2023). Treatment Trends and Combined Methods in Removing Pharmaceuticals and Personal Care Products from Wastewater—A Review. Membranes, 13(2). https://doi.org/10.3390/membranes13020158 Lolić, A., Paíga, P., Santos, L. H. M. L. M., Ramos, S., Correia, M., & Delerue-Matos, C. (2015). Assessment of non-steroidal anti-inflammatory and analgesic pharmaceuticals in seawaters of North of Portugal: Occurrence and environmental risk. Science of the Total Environment, 508, 240–250. https://doi.org/10.1016/j.scitotenv.2014.11.097 Lonappan, L., Rouissi, T., Kaur Brar, S., Verma, M., & Surampalli, R. Y. (2018). An insight into the adsorption of diclofenac on different biochars: Mechanisms, surface chemistry, and thermodynamics. Bioresource Technology, 249(October 2017), 386–394. https://doi.org/10.1016/j.biortech.2017.10.039 López-Vázquez, J., Pérez-Mayán, L., Fernández-Fernández, V., Cela, R., & Rodríguez, I. (2023). Direct, automated and sensitive determination of glyphosate and related anionic pesticides in environmental water samples using solid-phase extraction on-line combined with liquid chromatography tandem mass spectrometry. Journal of Chromatography A, 1687. https://doi.org/10.1016/j.chroma.2022.463697 Lopez, F. J., Pitarch, E., Botero-Coy, A. M., Fabregat-Safont, D., Ibáñez, M., Marin, J. M., Peruga, A., Ontañón, N., Martínez-Morcillo, S., Olalla, A., Valcárcel, Y., Varó, I., & Hernández, F. (2022). Removal efficiency for emerging contaminants in a WWTP from Madrid (Spain) after secondary and tertiary treatment and environmental impact on the Manzanares River. Science of the Total Environment, 812. https://doi.org/10.1016/j.scitotenv.2021.152567 Low, M. . J. D. (1960). Kinetics of Chemisorption of Gases on Solids. Chemical Reviews, 60(3), 267–312. https://doi.org/10.1021/cr60205a003 Lu, H. P., Li, Z. A., Gascó, G., Méndez, A., Shen, Y., & Paz-Ferreiro, J. (2018). Use of magnetic biochars for the immobilization of heavy metals in a multi-contaminated soil. Science of the Total Environment, 622–623, 892–899. https://doi.org/10.1016/j.scitotenv.2017.12.056 Lutri, V. F., Matteoda, E., Blarasin, M., Aparicio, V., Giacobone, D., Maldonado, L., Becher Quinodoz, F., Cabrera, A., & Giuliano Albo, J. (2020). Hydrogeological features affecting spatial distribution of glyphosate and AMPA in groundwater and surface water in an agroecosystem. Córdoba, Argentina. Science of the Total Environment, 711, 134557. https://doi.org/10.1016/j.scitotenv.2019.134557 Ma, L., Liu, Y., Yang, Q., Jiang, L., & Li, G. (2022). Occurrence and distribution of Pharmaceuticals and Personal Care Products (PPCPs) in wastewater related riverbank groundwater. Science of the Total Environment, 821(59), 153372. https://doi.org/10.1016/j.scitotenv.2022.153372 Mac Loughlin, T. M., Peluso, M. L., Aparicio, V. C., & Marino, D. J. G. (2020). Contribution of soluble and particulate-matter fractions to the total glyphosate and AMPA load in water bodies associated with horticulture. Science of the Total Environment, 703. https://doi.org/10.1016/j.scitotenv.2019.134717 References 234 Ortega-Calvo, J. J., Tejeda-Agredano, M. C., Jimenez-Sanchez, C., Congiu, E., Sungthong, R., Niqui-Arroyo, J. L., & Cantos, M. (2013). Is it possible to increase bioavailability but not environmental risk of PAHs in bioremediation? Journal of Hazardous Materials, 261, 733– 745. https://doi.org/10.1016/j.jhazmat.2013.03.042 Ortiz de García, S., Pinto Pinto, G., García Encina, P., & Irusta Mata, R. (2013). Consumption and occurrence of pharmaceutical and personal care products in the aquatic environment in Spain. Science of the Total Environment, 444, 451–465. https://doi.org/10.1016/j.scitotenv.2012.11.057 Ospina, M., Schütze, A., Morales-Agudelo, P., Vidal, M., Wong, L.-Y., & Calafat, A. M. (2022). Exposure to glyphosate in the United States: Data from the 2013–2014 National Health and Nutrition Examination Survey. Environment International, 170(November), 107620. https://doi.org/10.1016/j.envint.2022.107620 Othman, N. H., Alias, N. H., Fuzil, N. S., Marpani, F., Shahruddin, M. Z., Chew, C. M., Ng, K. M. D., Lau, W. J., & Ismail, A. F. (2022). A review on the use of membrane technology systems in developing countries. Membranes, 12(1). https://doi.org/10.3390/membranes12010030 Özçimen, D., & Ersoy-Meriçboyu, A. (2009). Removal of copper from aqueous solutions by adsorption onto chestnut shell and grapeseed activated carbons. Journal of Hazardous Materials, 168, 1118–1125. https://doi.org/10.1016/j.jhazmat.2009.02.148 Padilla, J. T., & Selim, H. M. (2019). Time‐Dependent Sorption and Desorption of Glyphosate in Soils: Multi‐reaction Modeling. Vadose Zone Journal, 18(1), 1–10. https://doi.org/10.2136/vzj2018.12.0214 Padilla, J. T., & Selim, H. M. (2020). Environmental behavior of glyphosate in soils. In Advances in Agronomy (1st ed., Vol. 159). Elsevier Inc. https://doi.org/10.1016/bs.agron.2019.07.005 Paíga, P., Santos, L. H. M. L. M., & Delerue-Matos, C. (2017). Development of a multi-residue method for the determination of human and veterinary pharmaceuticals and some of their metabolites in aqueous environmental matrices by SPE-UHPLC–MS/MS. Journal of Pharmaceutical and Biomedical Analysis, 135, 75–86. https://doi.org/10.1016/j.jpba.2016.12.013 Paíga, Paula, Correia, M., Fernandes, M. J., Silva, A., Carvalho, M., Vieira, J., Jorge, S., Silva, J. G., Freire, C., & Delerue-Matos, C. (2019). Assessment of 83 pharmaceuticals in WWTP influent and effluent samples by UHPLC-MS/MS: Hourly variation. Science of the Total Environment, 648(648), 582 – 600. https://doi.org/10.1016/j.scitotenv.2018.08.129 Paíga, Paula, Santos, L. H. M. L. M., Ramos, S., Jorge, S., Silva, J. G., & Delerue-Matos, C. (2016). Presence of pharmaceuticals in the Lis river (Portugal): Sources, fate and seasonal variation. Science of the Total Environment, 573, 164–177. https://doi.org/10.1016/j.scitotenv.2016.08.089 Pan, C., Zhu, F., Wu, M., Jiang, L., Zhao, X., & Yang, M. (2022). Degradation and toxicity of the antidepressant fluoxetine in an aqueous system by UV irradiation. Chemosphere, 287, 132434. https://doi.org/https://doi.org/10.1016/j.chemosphere.2021.132434 Maria João Cerveira Fernandes 235 Papageorgiou, M., Kosma, C., & Lambropoulou, D. (2016). Seasonal occurrence, removal, mass loading and environmental risk assessment of 55 pharmaceuticals and personal care products in a municipal wastewater treatment plant in Central Greece. Science of the Total Environment, 543, 547–569. https://doi.org/10.1016/j.scitotenv.2015.11.047 Patel, M., Kumar, R., Kishor, K., Mlsna, T., Pittman, C. U., & Mohan, D. (2019). Pharmaceuticals of emerging concern in aquatic systems: Chemistry, occurrence, effects, and removal methods. Chemical Reviews, 119(6), 3510–3673. https://doi.org/10.1021/acs.chemrev.8b00299 Pathak, V. M., Verma, V. K., Rawat, B. S., Kaur, B., Babu, N., Sharma, A., Dewali, S., Yadav, M., Kumari, R., Singh, S., Mohapatra, A., Pandey, V., Rana, N., & Cunill, J. M. (2022). Current status of pesticide effects on environment, human health and it’s eco-friendly management as bioremediation: A comprehensive review. Frontiers in Microbiology, 13(August), 1–29. https://doi.org/10.3389/fmicb.2022.962619 Pavlovic, D. M., Pinušić, T., Periša, M., & Babić, S. (2012). Optimization of matrix solid-phase dispersion for liquid chromatography tandem mass spectrometry analysis of 12 pharmaceuticals in sediments. Journal of Chromatography A, 1258, 1–15. https://doi.org/10.1016/j.chroma.2012.08.025 Pazikowska-Sapota, G., Galer-Tatarowicz, K., Dembska, G., Wojtkiewicz, M., Duljas, E., Pietrzak, S., & Dzierzbicka-Glowacka, L. A. (2020). The impact of pesticides used at the agricultural land of the Puck commune on the environment of the Puck Bay. PeerJ, 2020(3), 1–21. https://doi.org/10.7717/peerj.8789 Pazzagli, L., Reutfors, J., Lucian, E., Zerial, G., Perulli, A., & Castelpietra, G. (2022). Increased antidepressant use during the COVID-19 pandemic: Findings from the Friuli Venezia Giulia region, Italy, 2015–2020. Psychiatry Research, 315(June), 114704. https://doi.org/10.1016/j.psychres.2022.114704 Pereira, E., Melo, V. F., Abate, G., & Masini, J. C. (2019). Adsorption of glyphosate on Brazilian subtropical soils rich in iron and aluminum oxides. Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural Wastes, 54(11), 906–914. https://doi.org/10.1080/03601234.2019.1644947 Pereira, H. A., Hernandes, P. R. T., Netto, M. S., Reske, G. D., Vieceli, V., Oliveira, L. F. S., & Dotto, G. L. (2021). Adsorbents for glyphosate removal in contaminated waters: a review. Environmental Chemistry Letters, 19(2), 1525–1543. https://doi.org/10.1007/s10311- 020-01108-4 Perelo, L. W. (2010). Review: In situ and bioremediation of organic pollutants in aquatic sediments. Journal of Hazardous Materials, 177(1–3), 81–89. https://doi.org/10.1016/j.jhazmat.2009.12.090 Pérez-Fernández, V., Mainero Rocca, L., Tomai, P., Fanali, S., & Gentili, A. (2017). Recent advancements and future trends in environmental analysis: Sample preparation, liquid chromatography and mass spectrometry. Analytica Chimica Acta, 983, 9–41. https://doi.org/10.1016/j.aca.2017.06.029 Pérez-Parada, A., Goyenola, G., Mello, F. T. De, & Heinzen, H. (2018). Recent advances and References 236 open questions around pesticide dynamics and effects on freshwater fishes. Current Opinion in Environmental Science & Health, 4, 38–44. https://doi.org/10.1016/j.coesh.2018.08.004 Pérez, A. L., Tibaldo, G., Sánchez, G. H., Siano, G. G., Marsili, N. R., & Schenone, A. V. (2019). A novel fluorimetric method for glyphosate and AMPA determination with NBD-Cl and MCR-ALS. Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy, 214, 119–128. https://doi.org/10.1016/j.saa.2019.01.078 Pérez, D. J., Iturburu, F. G., Calderon, G., Oyesqui, L. A. E., De Gerónimo, E., & Aparicio, V. C. (2021). Ecological risk assessment of current-use pesticides and biocides in soils, sediments and surface water of a mixed land-use basin of the Pampas region, Argentina. Chemosphere, 263. https://doi.org/10.1016/j.chemosphere.2020.128061 Piccirillo, C., Moreira, I. S., Novais, R. M., Fernandes, A. J. S., Pullar, R. C., & Castro, P. M. L. (2017). Biphasic apatite-carbon materials derived from pyrolysed fish bones for effective adsorption of persistent pollutants and heavy metals. Journal of Environmental Chemical Engineering, 5(5), 4884–4894. https://doi.org/10.1016/j.jece.2017.09.010 Pinto, E., Soares, A. G., & Ferreira, I. M. P. L. V. O. (2018). Quantitative analysis of glyphosate, glufosinate and AMPA in irrigation water by: In situ derivatization-dispersive liquid-liquid microextraction combined with UPLC-MS/MS. Analytical Methods, 10(5), 554–561. https://doi.org/10.1039/c7ay02722b Pires, N. L., de Araújo, E. P., Oliveira-Filho, E. C., & Caldas, E. D. (2023). An ultrasensitive LCMS/MS method for the determination of glyphosate, AMPA and glufosinate in water – analysis of surface and groundwater from a hydrographic basin in the Midwestern region of Brazil. Science of The Total Environment, 875(December 2022), 162499. https://doi.org/10.1016/j.scitotenv.2023.162499 Pires, N. L., Passos, C. J. S., Morgado, M. G. A., Mello, D. C., Infante, C. M. C., & Caldas, E. D. (2020). Determination of glyphosate, AMPA and glufosinate by high performance liquid chromatography with fluorescence detection in waters of the Santarém Plateau, Brazilian Amazon. Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural Wastes, 55(9), 794–802. https://doi.org/10.1080/03601234.2020.1784668 Poiger, T., Buerge, I. J., Bächli, A., Müller, M. D., & Balmer, M. E. (2017). Occurrence of the herbicide glyphosate and its metabolite AMPA in surface waters in Switzerland determined with on-line solid phase extraction LC-MS/MS. Environmental Science and Pollution Research, 24(2), 1588–1596. https://doi.org/10.1007/s11356-016-7835-2 Pokharel, A., Acharya, B., & Farooque, A. (2020). Biochar-Assisted Wastewater Treatment and waste Valorization. In A. Abdelhafez & M. Abbas (Eds.), Applications of Biochar for Environmental Safety (p. 19). IntechOpen. https://doi.org/http://dx.doi.org/10.5772/intechopen.92288 Portail Substaces Chimiques. (2020). Normes de Qualité Environnementale et Valeurs Guides Environnementales. https://substances.ineris.fr/fr/substance/cas/1066-51-9/3 Portier, C. J. (2020). A comprehensive analysis of the animal carcinogenicity data for Maria João Cerveira Fernandes 237 glyphosate from chronic exposure rodent carcinogenicity studies. Environmental Health: A Global Access Science Source, 19(1), 1–18. https://doi.org/10.1186/s12940-020-00574- 1 Postigo, C., Ginebreda, A., Barbieri, M. V., Barceló, D., Martín-Alonso, J., de la Cal, A., Boleda, M. R., Otero, N., Carrey, R., Solà, V., Queralt, E., Isla, E., Casanovas, A., Frances, G., & López de Alda, M. (2021). Investigative monitoring of pesticide and nitrogen pollution sources in a complex multi-stressed catchment: The lower Llobregat River basin case study (Barcelona, Spain). Science of the Total Environment, 755, 142377. https://doi.org/10.1016/j.scitotenv.2020.142377 Pradhan, B., Chand, S., Chand, S., Rout, P. R., & Naik, S. K. (2023). Emerging groundwater contaminants: A comprehensive review on their health hazards and remediation technologies. Groundwater for Sustainable Development, 20(July 2022), 100868. https://doi.org/10.1016/j.gsd.2022.100868 Primost, J. E., Marino, D. J. G., Aparicio, V. C., Costa, J. L., & Carriquiriborde, P. (2017). Glyphosate and AMPA, “pseudo-persistent” pollutants under real-world agricultural management practices in the Mesopotamic Pampas agroecosystem, Argentina. Environmental Pollution, 229, 771–779. https://doi.org/10.1016/j.envpol.2017.06.006 Programa Nacional para a Saúde Mental. (2017). Programa Nacional para a Saúde Mental. Puri, M., Gandhi, K., & Kumar, M. S. (2023). Emerging environmental contaminants: A global perspective on policies and regulations. Journal of Environmental Management, 332(November 2022), 117344. https://doi.org/10.1016/j.jenvman.2023.117344 Qian, K., Kumar, A., Patil, K., Bellmer, D., Wang, D., Yuan, W., & Huhnke, R. L. (2013). Effects of biomass feedstocks and gasification conditions on the physiochemical properties of char. Energies, 6(8), 3972–3986. https://doi.org/10.3390/en6083972 Raby, M., Lissemore, L., Kaltenecker, G., Beaton, D., & Prosser, R. S. (2022). Characterizing the exposure of streams in southern Ontario to agricultural pesticides. Chemosphere, 294(November 2021), 133769. https://doi.org/10.1016/j.chemosphere.2022.133769 Rahman, F., Ismail, A., Yusof, S., Mazlan, N., & Engku-Ahmad-Khairi, E. A. (2019). Evaluation of glyphosate levels in sediments of milky stork foraging areas in Kuala Gula Bird Sanctuary, Perak, Malaysia. Pertanika Journal of Tropical Agricultural Science, 42(3), 995–1007. Raina-Fulton, R. (2014). A review of methods for the analysis of orphan and difficult pesticides: Glyphosate, glufosinate, quaternary ammonium and phenoxy acid herbicides, and dithiocarbamate and phthalimide fungicides. Journal of AOAC International, 97(4), 965– 977. https://doi.org/10.5740/jaoacint.SGERaina-Fulton Raj, A., Dubey, A., Ahmad, M., & Kumar, A. (2023). Pesticide pestilence : Global scenario and recent advances in detection and degradation methods. Journal of Environmental Management, 338(April), 117680. https://doi.org/10.1016/j.jenvman.2023.117680 Rajapaksha, A. U., Vithanage, M., Lim, J. E., Ahmed, M. B. M., Zhang, M., Lee, S. S., & Ok, Y. S. (2014). Invasive plant-derived biochar inhibits sulfamethazine uptake by lettuce in soil. Chemosphere, 111, 500–504. https://doi.org/10.1016/J.CHEMOSPHERE.2014.04.040 References 238 Ramirez-Haberkon, N. B., Aimar, S. B., Aparicio, V. C., Buschiazzo, D. E., De Gerónimo, E., Costa, J. L., & Mendez, M. J. (2021). Management effects on glyphosate and AMPA concentrations in the PM10 emitted by soils of the central semi-arid region of Argentina. Aeolian Research, 49(December 2020). https://doi.org/10.1016/j.aeolia.2020.100658 Ramirez-Haberkon, N. B., Aparicio, V. C., & Mendez, M. J. (2021). First evidence of glyphosate and aminomethylphosphonic acid (AMPA) in the respirable dust (PM10) emitted from unpaved rural roads of Argentina. Science of The Total Environment, 773, 145055. https://doi.org/10.1016/j.scitotenv.2021.145055 Ramirez, C. E., Bellmund, S., & Gardinali, P. R. (2014). A simple method for routine monitoring of glyphosate and its main metabolite in surface waters using lyophilization and LC-FLD + MS/MS. Case study: Canals with influence on Biscayne National Park. Science of the Total Environment, 496, 389–401. https://doi.org/10.1016/j.scitotenv.2014.06.118 Redshaw, C. H., Cooke, M. P., Talbot, H. M., McGrath, S., & Rowland, S. J. (2008). Low biodegradability of fluoxetine HCl, diazepam and their human metabolites in sewage sludge-amended soil. Journal of Soils and Sediments, 8(4), 217–230. https://doi.org/10.1007/s11368-008-0024-2 Reid, B. J., Jones, K. C., & Semple, K. T. (2000). Bioavailability of persistent organic pollutants in soils and sediments - A perspective on mechanisms, consequences and assessment. Environmental Pollution, 108(1), 103–112. https://doi.org/10.1016/S0269- 7491(99)00206-7 Reynoso, E. C., Peña, R. D., Reyes, D., Chavarin-Pineda, Y., Palchetti, I., & Torres, E. (2020). Determination of glyphosate in water from a rural locality in méxico and its implications for the population based on water consumption and use habits. International Journal of Environmental Research and Public Health, 17(19), 1–16. https://doi.org/10.3390/ijerph17197102 Rezaei Kalantary, R., Barzegar, G., & Jorfi, S. (2022). Monitoring of pesticides in surface water, pesticides removal efficiency in drinking water treatment plant and potential health risk to consumers using Monte Carlo simulation in Behbahan City, Iran. Chemosphere, 286(P1), 131667. https://doi.org/10.1016/j.chemosphere.2021.131667 Richards, B. K., Pacenka, S., Meyer, M. T., Dietze, J. E., Schatz, A. L., Teuffer, K., Aristilde, L., & Steenhuis, T. S. (2018). Antecedent and Post-Application Rain Events Trigger Glyphosate Transport from Runoff-Prone Soils. Environmental Science and Technology Letters, 5(5), 249–254. https://doi.org/10.1021/acs.estlett.8b00085 Rissouli, L., Benicha, M., Chafik, T., & Chabbi, M. (2017). Decontamination of water polluted with pesticide using biopolymers : Adsorption of glyphosate by chitin and chitosan. 8(12), 4544–4549. Rivas-Garcia, T., Espinosa-Calderón, A., Hernández-Vázquez, B., & Schwentesius-Rindermann, R. (2022). Overview of Environmental and Health Effects Related to Glyphosate Usage. Sustainability (Switzerland), 14(11), 1–14. https://doi.org/10.3390/su14116868 Rivera-Utrilla, J., Bautista-Toledo, I., Ferro-Garca, M. A., & Moreno-Castilla, C. (2001). Activated carbon surface modifications by adsorption of bacteria and their effect on Maria João Cerveira Fernandes 239 aqueous lead adsorption. Journal of Chemical Technology and Biotechnology, 76(12), 1209–1215. https://doi.org/10.1002/jctb.506 Rocío-Bautista, P., Moreno-González, D., Martínez-Piernas, A. B., García-Reyes, J. F., & Molina- Díaz, A. (2022). Novel liquid chromatography/mass spectrometry-based approaches for the determination of glyphosate and related compounds: A review. Trends in Environmental Analytical Chemistry, 36(e00186), 1–11. https://doi.org/10.1016/j.teac.2022.e00186 Rodrigues, M. do R., Lebre, A. I., Alves, A., Félix, A. M., Tavares, D., Noriega, E., Neves, I., Valente, M., Silva, M. G., Faria, M. J., Pereira, N., & Pacheco, P. (2018). Infeções e Resistências aos Antimicrobianos - Relatório Anual do Programa Prioritário 2018. Rodríguez-Bolaña, C., Pérez-Parada, A., Tesitore, G., Goyenola, G., Kröger, A., Pacheco, M., Gérez, N., Berton, A., Zinola, G., Gil, G., Mangarelli, A., Pequeño, F., Besil, N., Niell, S., Heinzen, H., & Teixeira de Mello, F. (2023). Multicompartmental monitoring of legacy and currently used pesticides in a subtropical lake used as a drinking water source (Laguna del Cisne, Uruguay). Science of the Total Environment, 874(February). https://doi.org/10.1016/j.scitotenv.2023.162310 Rodriguez-Mozaz, S., Ricart, M., Köck-Schulmeyer, M., Guasch, H., Bonnineau, C., Proia, L., de Alda, M. L., Sabater, S., & Barceló, D. (2015). Pharmaceuticals and pesticides in reclaimed water: Efficiency assessment of a microfiltration-reverse osmosis (MF-RO) pilot plant. Journal of Hazardous Materials, 282, 165–173. https://doi.org/10.1016/j.jhazmat.2014.09.015 Rosales, E., Meijide, J., Pazos, M., & Sanromán, M. A. (2017). Challenges and recent advances in biochar as low-cost biosorbent: From batch assays to continuous-flow systems. Bioresource Technology, 246, 176–192. https://doi.org/10.1016/j.biortech.2017.06.084 Rubio, R. S. (2018). Contribution to waste valorisation by producing biochars for different environmental purposes [Universidade de Santiago de Compostela]. https://dialnet.unirioja.es/servlet/tesis?codigo=171691 Ruiz, F., Cherubin, M. R., & Ferreira, T. O. (2020). Soil quality assessment of constructed Technosols: Towards the validation of a promising strategy for land reclamation, waste management and the recovery of soil functions. Journal of Environmental Management, 276(August). https://doi.org/10.1016/j.jenvman.2020.111344 S.K. Lagergren. (1898). About the Theory of So-called Adsorption of Soluble Substances. Sven. Vetenskapsakad. Handingarl, 24, 1–39. https://cir.nii.ac.jp/crid/1570009750361875328.bib?lang=en Saiz-Rubio, R., Balseiro-Romero, M., Antelo, J., Díez, E., Fiol, S., & Macías, F. (2019). Biochar as low-cost sorbent of volatile fuel organic compounds: potential application to water remediation. Environmental Science and Pollution Research, 26(12), 11605–11617. https://doi.org/10.1007/s11356-018-3798-9 Sakari, M., Hsia, B. S., Tahir, R., Rafiq, S., Nahavandi, R., Shah, M. D., Annammala, K. V., Soon, Z. Y., & Yi, A. L. J. (2014). Estuary and Sea - associated Wetlands as Final Sink for Organic Pollutants : A Case Study in Sabah , Malaysia. International Journal of Environment and References 240 Bioenergy, 9(1), 1–16. https://doi.org/10.1111/jocn.13970 Samal, K., Mahapatra, S., & Hibzur Ali, M. (2022). Pharmaceutical wastewater as Emerging Contaminants (EC): Treatment technologies, impact on environment and human health. Energy Nexus, 6(February), 100076. https://doi.org/10.1016/j.nexus.2022.100076 Samson-Brais, É., Lucotte, M., Moingt, M., Tremblay, G., & Paquet, S. (2022). Glyphosate and aminomethylphosphonic acid contents in field crops soils under various weed management practices. Agrosystems, Geosciences and Environment, 5(3), 1–15. https://doi.org/10.1002/agg2.20273 Santos, E., Abreu, M. M., Macías, F., Anjos, C., Magalhães, M. C., & de Varennes, A. (2010). Efeito de correctivos orgânicos e inorgânicos na recuperação de escombreiras ricas em sulfuretos. In D. Flores & M. Marques (Eds.), X Congresso de Geoquímica dos Países de Língua Portuguesa - XVI Semana Geoquímica (pp. 1059–1065). Santos, E. S., Abreu, M. M., & Macías, F. (2019). Rehabilitation of mining areas through integrated biotechnological approach: Technosols derived from organic/inorganic wastes and autochthonous plant development. Chemosphere, 224, 765–775. https://doi.org/10.1016/j.chemosphere.2019.02.172 Santos, E. S., Abreu, M. M., Macías, F., & Magalhães, M. C. F. (2017). Potential environmental impact of technosols composed of gossan and sulfide-rich wastes from São Domingos mine: assay of simulated leaching. Journal of Soils and Sediments, 17(5), 1369–1383. https://doi.org/10.1007/s11368-016-1518-y Santos, J. L., Martín, J., Mejías, C., Aparicio, I., & Alonso, E. (2022). Pharmaceuticals and Their Metabolites in Sewage Sludge and Soils: Distribution and Environmental Risk Assessment. In A. Núñez-Delgado & M. Arias-Estévez (Eds.), Emerging Pollutants in Sewage Sludge and Soils. The Handbook of Environmental Chemistry, vol. 114 (pp. 19–36). Springer International Publishing. https://doi.org/10.1007/698_2022_847 Santos, L. H. M. L. M., Ramalhosa, M. J., Ferreira, M., & Delerue-Matos, C. (2016). Development of a modified acetonitrile-based extraction procedure followed by ultrahigh performance liquid chromatography-tandem mass spectrometry for the analysis of psychiatric drugs in sediments. Journal of Chromatography A, 1437, 37–48. https://doi.org/10.1016/j.chroma.2016.01.079 Sardella, F., Gimenez, M., Navas, C., Morandi, C., Deiana, C., & Sapag, K. (2015). Conversion of viticultural industry wastes into activated carbons for removal of lead and cadmium. Journal of Environmental Chemical Engineering, 3(1), 253–260. https://doi.org/10.1016/j.jece.2014.06.026 Sarıkaya, M., Ulusoy, H. I., Morgul, U., Ulusoy, S., Tartaglia, A., Yılmaz, E., Soylak, M., Locatelli, M., & Kabir, A. (2021). Sensitive determination of Fluoxetine and Citalopram antidepressants in urine and wastewater samples by liquid chromatography coupled with photodiode array detector. Journal of Chromatography A, 1648, 462215. https://doi.org/https://doi.org/10.1016/j.chroma.2021.462215 Sasal, M. C., Wilson, M. G., Sione, S. M., Beghetto, S. M., Gabioud, E. A., Oszust, J. D., Paravani, E. V., Demonte, L., Repetti, M. R., Bedendo, D. J., Medero, S. L., Goette, J. J., Pautasso, N., Maria João Cerveira Fernandes 241 & Schulz, G. A. (2017). Monitoreo de glifosato en agua superficial en Entre Ríos. La investigación acción participativa como metodología de abordaje. Revista de Investigaciones Agropecuarias, 43(2), 195–205. Sayğılı, H., Güzel, F., & Önal, Y. (2015). Conversion of grape industrial processing waste to activated carbon sorbent and its performance in cationic and anionic dyes adsorption. Journal of Cleaner Production, 93, 84–93. https://doi.org/10.1016/j.jclepro.2015.01.009 Schleiffer, M., & Speiser, B. (2022). Presence of pesticides in the environment, transition into organic food, and implications for quality assurance along the European organic food chain – A review. Environmental Pollution, 313(August), 120116. https://doi.org/10.1016/j.envpol.2022.120116 Schütze, A., Morales-Agudelo, P., Vidal, M., Calafat, A. M., & Ospina, M. (2021). Quantification of glyphosate and other organophosphorus compounds in human urine via ion chromatography isotope dilution tandem mass spectrometry. Chemosphere, 274, 129427. https://doi.org/10.1016/j.chemosphere.2020.129427 Seesen, M., & Sapbamrer, R. (2020). A systematic review of association between pesticide exposure and respiratory outcomes among farmers and farmworkers. Malaysian Journal of Medicine and Health Sciences, 16(1), 312–324. Séralini, G. E., Clair, E., Mesnage, R., Gress, S., Defarge, N., Malatesta, M., Hennequin, D., & de Vendômois, J. S. (2014). Republished study: long-term toxicity of a Roundup herbicide and a Roundup-tolerant genetically modified maize. Environmental Sciences Europe, 26(1), 1–17. https://doi.org/10.1186/s12302-014-0014-5 Sharma, A. D., & Lai, D. (2019). Sorption of radiolabelled glyphosate on biochar aged in contrasting soils. Journal of Environmental Science and Health - Part B Pesticides, Food Contaminants, and Agricultural Wastes, 54(1), 49–53. https://doi.org/10.1080/03601234.2018.1531658 Sharma, Akanksha, Shukla, A., Attri, K., Kumar, M., Kumar, P., Suttee, A., Singh, G., Barnwal, R. P., & Singla, N. (2020). Global trends in pesticides: A looming threat and viable alternatives. Ecotoxicology and Environmental Safety, 201(110812), 1–15. https://doi.org/10.1016/j.ecoenv.2020.110812 Sharma, Anket, Kumar, V., Shahzad, B., Tanveer, M., Sidhu, G. P. S., Handa, N., Kohli, S. K., Yadav, P., Bali, A. S., Parihar, R. D., Dar, O. I., Singh, K., Jasrotia, S., Bakshi, P., Ramakrishnan, M., Kumar, S., Bhardwaj, R., & Thukral, A. K. (2019). Worldwide pesticide usage and its impacts on ecosystem. SN Applied Sciences, 1(11), 1–16. https://doi.org/10.1007/s42452-019-1485-1 Sharma, B. M., Bečanová, J., Scheringer, M., Sharma, A., Bharat, G. K., Whitehead, P. G., Klánová, J., & Nizzetto, L. (2019). Health and ecological risk assessment of emerging contaminants (pharmaceuticals, personal care products, and artificial sweeteners) in surface and groundwater (drinking water) in the Ganges River Basin, India. Science of the Total Environment, 646, 1459–1467. https://doi.org/10.1016/j.scitotenv.2018.07.235 Sharma, J., Joshi, M., Bhatnagar, A., Chaurasia, A. K., & Nigam, S. (2022). Pharmaceutical residues: One of the significant problems in achieving ‘clean water for all’ and its solution. References 242 Environmental Research, 215(P1), 114219. https://doi.org/10.1016/j.envres.2022.114219 Sidoli, P., Baran, N., & Angulo-Jaramillo, R. (2016). Glyphosate and AMPA adsorption in soils: laboratory experiments and pedotransfer rules. Environmental Science and Pollution Research, 23(6), 5733–5742. https://doi.org/10.1007/s11356-015-5796-5 Silori, R., Shrivastava, V., Singh, A., Sharma, P., Aouad, M., Mahlknecht, J., & Kumar, M. (2022). Global groundwater vulnerability for Pharmaceutical and Personal care products (PPCPs): The scenario of second decade of 21st century. Journal of Environmental Management, 320(115703), 1–18. https://doi.org/10.1016/j.jenvman.2022.115703 Silva-Madera, R. J., Salazar-Flores, J., Peregrina-Lucano, A. A., Mendoza-Michel, J., Ceja- Gálvez, H. R., Rojas-Bravo, D., Reyna-Villela, M. Z., & Torres-Sánchez, E. D. (2021). Pesticide Contamination in Drinking and Surface Water in the Cienega, Jalisco, Mexico. Water, Air, and Soil Pollution, 232(2). https://doi.org/10.1007/s11270-021-04990-y Silva, Andreia, Fernandes, D. F., Figueiredo, S. A., Freitas, O. M., & Delerue-Matos, C. (2022). Fluoxetine and Nutrients Removal from Aqueous Solutions by Phycoremediation. International Journal of Environmental Research and Public Health, 19(10). https://doi.org/10.3390/ijerph19106081 Silva, Andreia, Martinho, S., Stawiński, W., Węgrzyn, A., Figueiredo, S., Santos, L. H. M. L. M., & Freitas, O. (2018). Application of vermiculite-derived sustainable adsorbents for removal of venlafaxine. Environmental Science and Pollution Research, 25(17), 17066– 17076. https://doi.org/10.1007/s11356-018-1869-6 Silva, Andreia, Stawiński, W., Romacho, J., Santos, L. H. M. L. M., Figueiredo, S. A., Freitas, O. M., & Delerue-Matos, C. (2019). Adsorption of Fluoxetine and Venlafaxine onto the Marine Seaweed Bifurcaria bifurcata. Environmental Engineering Science, 36(5), 573– 582. https://doi.org/10.1089/ees.2018.0332 Silva, Aurora, Santos, L. H. M. L. M., Delerue-Matos, C., & Figueiredo, S. A. (2014). Impact of excipients in the chronic toxicity of fluoxetine on the alga Chlorella vulgaris. Environmental Technology (United Kingdom), 35(24), 3124–3129. https://doi.org/10.1080/09593330.2014.932438 Silva, V., Mol, H. G. J., Zomer, P., Tienstra, M., Ritsema, C. J., & Geissen, V. (2019). Pesticide residues in European agricultural soils – A hidden reality unfolded. Science of the Total Environment, 653, 1532–1545. https://doi.org/10.1016/j.scitotenv.2018.10.441 Silva, V., Montanarella, L., Jones, A., Fernández-Ugalde, O., Mol, H. G. J. J., Ritsema, C. J., & Geissen, V. (2018). Distribution of glyphosate and aminomethylphosphonic acid (AMPA) in agricultural topsoils of the European Union. Science of the Total Environment, 621, 1352–1359. https://doi.org/10.1016/j.scitotenv.2017.10.093 Singh, S., Kumar, V., Gill, J. P. K., Datta, S., Singh, S., Dhaka, V., Kapoor, D., Wani, A. B., Dhanjal, D. S., Kumar, M., Harikumar, S. L., & Singh, J. (2020). Herbicide glyphosate: Toxicity and microbial degradation. International Journal of Environmental Research and Public Health, 17(20), 1–18. https://doi.org/10.3390/ijerph17207519 Skark, C., Zullei-Seibert, N., Schöttler, U., & Schlett, C. (1998). The occurrence of glyphosate in Maria João Cerveira Fernandes 243 surface water. International Journal of Environmental Analytical Chemistry, 70(1–4), 93– 104. https://doi.org/10.1080/03067319808032607 Skeff, W., Recknagel, C., Düwel, Y., & Schulz-Bull, D. E. (2018). Adsorption behaviors of glyphosate, glufosinate, aminomethylphosphonic acid, and 2-aminoethylphosphonic acid on three typical Baltic Sea sediments. Marine Chemistry, 198(June 2017), 1–9. https://doi.org/10.1016/j.marchem.2017.11.008 Smith, N. J., Martin, R. C., & St. Croix, R. G. (1996). Levels of the herbicide glyphosate in well water. Bulletin of Environmental Contamination and Toxicology, 57(5), 759–765. https://doi.org/10.1007/s001289900254 Soares, S., Rosado, T., Barroso, M., & Gallardo, E. (2021). New method for the monitoring of antidepressants in oral fluid using dried spot sampling. Pharmaceuticals, 14(12), 1–20. https://doi.org/10.3390/ph14121284 Sohel, A. J., Shutter, M. C., & Molla, M. (2023). Fluoxetine. In StatPearls [Internet]. Solomon, K. R. (2020). Estimated exposure to glyphosate in humans via environmental, occupational, and dietary pathways: an updated review of the scientific literature. Pest Management Science, 76(9), 2878–2885. https://doi.org/10.1002/ps.5717 Sousa, J. C. G., Ribeiro, A. R., Barbosa, M. O., Pereira, M. F. R., & Silva, A. M. T. (2018). A review on environmental monitoring of water organic pollutants identified by EU guidelines. Journal of Hazardous Materials, 344, 146–162. https://doi.org/10.1016/j.jhazmat.2017.09.058 Souza, F. L., Saéz, C., Llanos, J., Lanza, M. R. V., Cañizares, P., & Rodrigo, M. A. (2016). Solarpowered electrokinetic remediation for the treatment of soil polluted with the herbicide 2,4-D. Electrochimica Acta, 190, 371–377. https://doi.org/10.1016/j.electacta.2015.12.134 Spengler, S. R., Heskett, M. D., & Spengler, S. C. (2021). Occurrence and Persistence of the Herbicide Glyphosate in a suburban tropical watershed. International Journal of Environmental Impacts: Management, Mitigation and Recovery, 4(2), 141–156. https://doi.org/10.2495/ei-v4-n2-141-156 Suárez, R. P., Goijman, A. P., Cappelletti, S., Solari, L. M., Cristos, D., Rojas, D., Krug, P., Babbitt, K. J., & Gavier-Pizarro, G. I. (2021). Combined effects of agrochemical contamination and forest loss on anuran diversity in agroecosystems of east-central Argentina. Science of the Total Environment, 759, 143435. https://doi.org/10.1016/j.scitotenv.2020.143435 Subsecretaria de Recursos Hídricos de la Nación. (2003). Desarrollos de niveles guia nacionales de calidad de agua ambiente correspondientes a difenilos policlorados. Niveles Guía Nacionales de Calidad de Agua Ambiente. Suciu, N., Russo, E., Calliera, M., Luciani, G. P., Trevisan, M., & Capri, E. (2023). Glyphosate, glufosinate ammonium, and AMPA occurrences and sources in groundwater of hilly vineyards. Science of the Total Environment, 866(161171), 1–9. https://doi.org/10.1016/j.scitotenv.2022.161171 Sun, M., Li, H., & Jaisi, D. P. (2019). Degradation of glyphosate and bioavailability of List of Publications 250 LIST OF PUBLICATIONS Published articles used in this Thesis: rights, contributions and quality indicators Antibiotics and antidepressants occurrence in surface waters and sediments collected in the north of Portugal Maria João Fernandesa,b, Paula Paígaa, Ana Silvaa, Carmen Pérez Llagunob, Manuela Carvalhoa, Felipe Macías Vázquezb, Cristina Delerue-Matosa aREQUIMTE/LAQV, Instituto Superior de Engenharia do Porto, Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 431, 4200-072 Porto, Portugal; bIIT/LTA - Instituto de Investigaciones Tecnológicas, Universidad de Santiago de Compostela, E-15782 Santiago de Compostela, Spain Chemosphere, ISSN: 0045-6535, volume 239 (2020): 124729, pages 1-12 DOI: 10.1016/j.chemosphere.2019.124729 Specific contribution to the publication Maria João Fernandes contributes with sample collection and pre-treatment, experimental work, namely: extraction and analysis, data statistical analysis and to the writing of the original article draft. Quality indicator: The journal Chemosphere, published by Elsevier Ltd., currently presents an impact factor of 8.943 (2022 Journal Citation Reports), an H-index of 265 and a CiteScore index (2021) of 11.7 (consulted in Scopus 31/03/2023). Is a journal covering the technologies/fields/categories related to Chemistry (miscellaneous) (Q1); Environmental Maria João Cerveira Fernandes 251 Chemistry (Q1); Environmental Engineering (Q1); Health, Toxicology and Mutagenesis (Q1); Medicine (miscellaneous) (Q1); Pollution (Q1); Public Health, Environmental and Occupational Health (Q1). The content of this journal article was entirely used in the sub-chapter 3.1.1. of this doctoral thesis, under the consent of the publisher (Elsevier). Evaluation of the adsorption potential of biochars prepared from forest and agri-food wastes for the removal of fluoxetine Maria João Fernandesa,b , Manuela M. Moreiraa,⁎ , Paula Paígaa , Diogo Diasc , Maria Bernardoc, Manuela Carvalhoa , Nuno Lapac , Isabel Fonsecac , Simone Moraisa , Sónia Figueiredoa , Cristina Delerue-Matosa aREQUIMTE/LAQV, Instituto Superior de Engenharia do Porto, Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 431, 4200-072 Porto, Portugal; List of Publications 252 bIIT/LTA - Instituto de Investigaciones Tecnológicas, Universidad de Santiago de Compostela, E-15782 Santiago de Compostela, Spain cLAQV/REQUIMTE, Faculdade de Ciências e Tecnologia (FCT), Universidade Nova de Lisboa (UNL), 2829-516 Caparica, Portugal Bioresource Technology, ISSN: 0045-6535, volume 292 (2019): 121973, pages 1-10 DOI: 10.1016/j.biortech.2019.121973 Specific contribution to the publication Maria João Fernandes contributes with sample pre-treatment, experimental work, namely: adsorption (batch and column) assays, and fluoxetine analysis, result discussion and the writing of the original article draft. Quality indicator: The journal Bioresource Technology, published by Elsevier Ltd., currently presents an impact factor of 11.889 (2021 Journal Citation Reports), an H-index of 341 and a CiteScore index (2021) of 17.4 (consulted in Scopus 31/03/2023). Is a journal its positions in the different categories according to Scimago are Bioengineering (Q1); Environmental Engineering (Q1); Waste Management and Disposal (Q1); Medicine (miscellaneous) and (Q1); Renewable Energy, Sustainability and the Environment (Q1). The content of this journal article was entirely used in in the chapter 4 of this doctoral thesis, under the consent of the publisher (Elsevier). Maria João Cerveira Fernandes 253 Other publications Peer Reviewed Journals Paula Paíga; Manuela Correia; Maria João Fernandes; Ana Silva; Manuela Carvalho; Joana Vieira; Sandra Jorge; Jaime G. Silva; Cristina Delerue-Matos. "Assessment of 83 pharmaceuticals in WWTP influent and effluent samples by UHPLC-MS/MS: Hourly variation". Science of The Total Environment 648 (2019): 582-600. http://www.sciencedirect.com/science/article/pii/S0048969718330936. Congress Proceedings Oral contributions Maria J. Fernandes, Paula Paíga, Sónia Figueiredo, Felipe Macías & Cristina Delerue-Matos. “ lyphosate and Aminomethylphosphonic acid adsorption in soils”. Presented in Congreso Internacional XXVI Encontro Galego Portugês de Química, 2022 Maria J. Fernandes, Felipe Macías & Cristina Delerue-Matos. “Tecnosolos a la carte: Um método inovador, abrangente e sustentável de remedição de contaminações de solos e águas”. Presented in GEO em FOCO, 2019 Poster contributions Maria João Fernandes; Paula Paíga; Ana Silva; Carmen Llaguno; Manuela Carvalho; Felipe Macías Vázquez; Cristina Delerue-Matos. "Assessment of antibiotics and psychiatric drugs in sediments from Douro and Leça Rivers (Portugal)". Presented in 4GEO|resources, materials, technologies & environment| 1st Conference 2019, 2019. Maria João Fernandes; Paula Paíga; Ana Silva; C. P. Llaguno; Manuela Carvalho; Felipe Macías Vázquez; Delerue-Matos, Cristina. "Determinación de fármacos psiquiátricos y antibióticos en las aguas superficiales de los ríos Douro y Leça (Portugal)". Presented in 1º Simposio "Presencia y eliminación de microcontaminantes en agua" de la Red_Novedar, 2019. Maria João Fernandes; Manuela Moreira; Diogo Dias; Maria Bernardo; Manuela Carvalho; Nuno Lapa; Isabel Fonseca; et al. "Micropollutant Removal From Water Through Biochar Adsorbents". Presented in 1st International Conference on Advanced Production and Processing (ICAPP), poster NP-P.52, Novi Sad, Sérvia, Novi Sad, 2019. Maria João Fernandes; Manuela M. Moreira; Paula Paíga; Silvia Capelas; Manuela Carvalho; Sónia Figueiredo; Simone Morais; Cristina Delerue-Matos. "Evaluation of the adsorption potential of biochars prepared from agro-food wastes for the removal of fluoxetine". Presented in XXIV Encontro Luso-Galego de Química, 2018. Fernandes, Maria; Silva, A.; Paíga, P.; Carvalho, M. M.; Llaguno, C.; Vázquez, F. Macías; Delerue-Matos, C.. "Determination of Glyphosate in Sediments: A New Analytical Approach". Presented in 40th International Conference on Environmental & Food Monitoring, Santiago de Compostela, 2018. List of Publications 254 Silva, A.; Maria João Fernandes; Paula Paíga; Carvalho, M. M.; Llaguno, C.; Vázquez, F. Macías; Delerue-Matos, C.. "Development of a QuEChERS Sample Preparation Method for Fluoxetine Determination in Sediments". Presented in 40th International Conference on Environmental & Food Monitoring, 2018. Maria João Cerveira Fernandes 255 ANNEX - AUTHORIZATION FOR USE AND ADAPTATION OF FIGURES Figure 2.1 and Figure 2.2 From the report: WHO/UNICEF. (2021). Progress on household drinking water, sanitation and hygiene 2000-2020: five years into the SDGs. Licence: CC BY-NC-SA 3.0 IGO. http://apps.who.int/bookorders. Use and adaptation authorization: Annex - Authorization For Use And Adaptation Of Figures 256 Figure 2.5 From: Silori, R., Shrivastava, V., Singh, A., Sharma, P., Aouad, M., Mahlknecht, J., & Kumar, M. (2022). Global groundwater vulnerability for Pharmaceutical and Personal care products (PPCPs): The scenario of second decade of 21st century. Journal of Environmental Management, 320(115703), 1–18. https://doi.org/10.1016/j.jenvman.2022.115703 Use and adaptation authorization: Maria João Cerveira Fernandes 257 Annex - Authorization For Use And Adaptation Of Figures 258 Maria João Cerveira Fernandes 259 Figure 2.15 From: Espinoza-Montero, P. J., Vega-Verduga, C., Alulema-Pullupaxi, P., Fernández, L., & Paz, J. L. (2020). Technologies employed in the treatment of water contaminated with glyphosate: A review. Molecules, 25(23), 1–26. https://doi.org/10.3390/molecules25235550, Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). Use and adaptation authorization: