Co esponding au ho : Ra ind a B. Malabadi
Copy igh © 2025 Au ho (s) e ain he copy igh o his a icle. This a icle is published unde he e ms o he C ea i e Commons A ibu ion Liscense 4.0.
Mic oplas ics: De ec ion me hods-An upda e
Raju K. Chalanna a 1, A inash A. Kamble 4, Ra ind a B. Malabadi 1, 2, *, Di aka MS 3, Swa hi 1, Kisho e S.
Ka amchand 5, Ki an P. Kolka 6, Somayyeh Mo amazi 7, An onia Neidilê Ribei o Munhoz 8, and Ka en Vi iana
Cas año Co onado 9
1 Depa men o Applied Bo any, Mangalo e Uni e si y, Mangalagango i-574199, Mangalo e, Ka na aka S a e, India.
2 Mille Bl d, NW, Edmon on, Albe a, Canada.
3 Food Science and Nu i ion, Depa men o Biosciences, Mangalo e Uni e si y, Mangalagango i- 574199, Ka na aka
S a e, India.
4 Depa men o Indus ial Chemis y, Mangalo e Uni e si y, Mangalagango i- 574199, Ka na aka S a e, India.
5 Depa men o Zoology, Poo nap ajna College, Au onomous, Udupi- 576101, Ka na aka S a e, India.
6 Depa men o Bo any, Ka na ak Science College, Dha wad-580003, Ka na aka S a e, India.
7 Depa men o Ho icul u e, Ag onomy, Science and Resea ch B anch, Islamic Azad Uni e si y, Teh an-1477893855, I an.
8 Depa men o Chemis y, En i onmen and Food, Fede al Ins i u e o Amazonas, Campus Manaus Cen o, Amazonas,
B azil- 69020-120.
9 Chie Communica ions O ice (CCO), Resea ch Issues and CO-Founde o LAIHA (La in Ame ican Indus ial Hemp
Associa ion), and CEO- CANNACONS, Bogo a, D.C., Capi al Dis ic , Colombia.
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2809-2824
Publica ion his o y: Recei ed on 10 Ma ch 2025; e ised on 20 Ap il 2025; accep ed on 10 May 2025
A icle DOI: h ps://doi.o g/10.30574/wja .2025.26.2.1715
Abs ac
Mic oplas ics a e syn he ic polyme s wi h majo dimension o ≤5 mm. The pa icles occu in a la ge a ie y o shapes,
sizes, colo s, and composi ions. Mic oplas ics en e he ood chain, hey may be biomagni ied and bioaccumula ed by
la ge o ganisms and ul ima ely each humans. Apa om o ganisms, o he ood ma e ials such as sal , honey, bee ,
ea bags, and d inking wa e ha e also been epo ed o ha e mic oplas ic con amina ion. O gans epo ed o be
con amina ed by mic oplas ics and nanoplas ics include he gas oin es inal ac , espi a o y sys em, skin, li e ,
kidneys, and e en he b ain. E ec o mic oplas ic con amina ion on hese o gans can ange om in lamma o y
esponses o issue damage and po en ial dis up ion o o gan unc ion and ca cinogenesis. Mic oplas ics ha e en e ed
d inking wa e ia a ious pa hways, aising conce ns abou hei po en ial heal h impac s. A numbe o luo escen
dyes, including Nile ed, Rhodamine B, Sa anin T, and luo escein iso phospha e, can label plas ic polyme s and hence
a e used in he de ec ion o mic oplas ics. Among hese, Nile ed has been used widely as a apid me hod o de ec ing
mic oplas ics. The h ee main me hods o de ec ing and quan i ying mic oplas ic concen a ions in wa e a e FTIR
Spec oscopy, py-GC/MS, and Raman Spec oscopy. FTIR and Raman Spec oscopy can de e mine he numbe o
mic oplas ic pa icles by plas ic ype and size ange, whe eas py-GC/MS can quan i y concen a ions o speci ic ypes o
mic oplas ics in mg/l. To o e come he challenges o ime and labo -in ensi e, mic oplas ic de ec ion echniques,
esea che s a e inc easingly adop ing machine lea ning and au oma ion. These echnologies can p ocess la ge da ase s
wi h g ea e speed and accu acy, aining algo i hms o de ec mic oplas ic mo e e icien ly.
Keywo ds: Fluo escen dyes; FTIR; Mic oplas ic; Machine lea ning; Mic oplas ic; Nile Red (NR); Raman Spec oscopy
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2809-2824
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1. In oduc ion
Plas ics we e i s de eloped in he ea ly 1900s. A e 1945 hey became mo e widely used, wi h a esul an d ama ic
inc ease in plas ic pollu ion and hei b eakdown o mic oplas ics. Global plas ic p oduc ion has almos doubled
compa ed o wo decades ago and mos o he plas ics a e ending up in land ill, incine a ed, o leaking in o he
en i onmen , wi h only 9% success ully ecycled globally [1-141-172]. The e m "mic oplas ics" (MPs) was coined in
2004 by P o esso Richa d Thompson o desc ibe small plas ic pa icles ound in a ious en i onmen s [7-9-141. 168,
169]. In de eloped coun ies, some plas ics a e ecycled, whe eas in low-income coun ies, no ad anced equipmen o
ecycling exis s. Mic oplas ics can be ound in a ious en i onmen al compa men s such as oceans, i e s, lakes, soil,
ai , and e en in o ganisms. Acco ding o ecen esea ch, mic oplas ics we e ound in la ge concen a ions in a ious
ood and be e age sou ces and we e de ec ed in human aeces. Mic oplas ics a e syn he ic polyme s wi h majo
dimension o ≤5 mm [1-141-166-172]. The pa icles occu in a la ge a ie y o shapes, sizes, colo s, and composi ions.
Poly Vinyl Chlo ide (PVC), Polye hylene Te eph hala e (PET), Polys y ene (PS), and Polye hylene (PE) a e he mos
abundan mic oplas ic polyme s ound in na u e, and gas oin es inal inges ion has been iden i ied as he p ima y
pa hway o exposu e o mic oplas ic [1-141-172]. Mic oplas ics, pe asi e en i onmen al con aminan s, pose a
po en ial isk o human heal h, including he de elopmen o o al ca cinoma [15, 17-141-172]. These minu e plas ic
pa icles in il a e a ious en i onmen al niches and en e he human body h ough inges ion, inhala ion, and de mal
exposu e [15, 17-141]. De ec ed in o gans such as he gas oin es inal ac and espi a o y sys em, mic oplas ics may
induce in lamma ion and o gan dys unc ion [15, 17-141-172]. Eme ging esea ch sugges s hei po en ial o ha bo
ca cinogenic subs ances, leading o DNA damage and ca cinogenesis [15, 17-141-169-172]. O e all, he e a e h ee
ou es o mic oplas ic elease in o he en i onmen – (i) di ec discha ge o plas ic was e in o wa e bodies; (ii)
discha ge om indus ial and domes ic was ewa e ea men plan s (WWTPs); (iii) plas ic was e in ishing essels,
ishe ies, and wa e ope a ion indus ies.
On he basis o li e a u e su ey, i is likely ha global con amina ion o mic oplas ics will be e en ually b ough back
o ou dinne able h ough consump ion o a ious ood i ems. Al hough a ew s udies ha e quan i a i ely es ima ed
he mic oplas ic consump ion o people om con amina ed sea ood , sal , and packaging ma e ials , he ex en o
people’s mic oplas ic exposu e ia ood consump ion emains la gely unknown [1-141-172] Once mic oplas ics en e
he ood chain, hey may be biomagni ied and bioaccumula ed by la ge o ganisms and ul ima ely each humans [4-15-
141]. Apa om o ganisms, o he ood ma e ials such as sal , honey, bee , ea bags, and d inking wa e ha e also been
epo ed o ha e mic oplas ic con amina ion [5-141]. These p oduc s a e egula ly used by humans and se e as
sou ces o he en y o mic oplas ics in o he human body [1-141-172] Inges ion occu s p ima ily h ough
con amina ed ood and wa e , inhala ion h ough ai con amina ed wi h mic oplas ic pa icles, and de mal exposu e
h ough di ec con ac wi h p oduc s con aining mic oplas ics o con amina ed su aces [1-15-141-172]. O gans
epo ed o be con amina ed by mic oplas ics and nanoplas ics include he gas oin es inal ac , espi a o y sys em,
li e , kidneys, and e en b ain [7-15]. E ec s o mic oplas ic con amina ion on hese o gans may include in lamma ion,
oxida i e s ess, and dis up ion o cellula unc ion [15, 17-141]. O gans epo ed o be con amina ed by mic oplas ics
and nanoplas ics include he gas oin es inal ac , espi a o y sys em, skin, li e , kidneys, and e en he b ain [8-17-
141]. E ec s o mic oplas ic con amina ion on hese o gans can ange om in lamma o y esponses o issue damage
and po en ial dis up ion o o gan unc ion [15, 17-141]. Mic o- and nanoplas ics can se e as a sou ce o ca cinogenic
o mu agenic subs ances, po en ially causing DNA damage ha can lead o ca cinogenesis, he de elopmen o
cance ous umo s [13-17-141]. Geno oxici y s udies in es iga ed he e ec s o mic oplas ics and associa ed chemicals
on DNA in eg i y, ch omosomal s uc u e, and genomic s abili y using sensi i e assays [15, 17-141-172]. Due o he
po en ial heal h isks and nega i e impac s on he en i onmen , he use o mic oplas ics in ea bags and o he p oduc s
is a majo issue[1-13-17-141]. Li e al., (2023) [64] epo ed ha mic oplas ics a e consumed by humans om an ea ly
age and in inc easingly la ge quan i ies [12-17]. As mic oplas ics pass h ough he gas oin es inal ac hey in e ac
wi h he no mal physiological mechanism o he body, pa icula ly in he colon and ec um, whe e hey may in e ac
wi h he p o ec i e colonic mucus laye [12-17]. Li e al., (2023) [64] epo ed se e al possible mechanisms o how
mic oplas ics may dis up his mucus laye , hus educing i s p o ec i e e ec and inc easing he likelihood o colo ec al
cance [12, 17-141-169]. The e o e, i is possible ha he mic oplas ics damage he ba ie in eg i y o he colonic
mucus laye , hus educing i s p o ec i e e ec [64]. Li e al., (2023) [64] also indica ed ha u he cla i ica ion needs
o be sough ega ding he in e ac ion be ween mic oplas ic, gu mic obio a and he mucus laye [15]. This will need o
be modeled in long- e m animal s udies o be e unde s and how ch onic consump ion o en i onmen ally-acqui ed
mic oplas ics may con ibu e o an inc eased isk o colo ec al ca cinogenesis [12-17-141-172].
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2. Mic oplas ics: Food
Now a days mos o he ood is con amina ed wi h mic o plas ics. Some o he common ood con aining mic oplas ics
a e able sal , suga , he bal medicine, ea bags, honey, bee and milk [1-141-172]. Because able sal is mos o en
p oduced by he dis illa ion o seawa e , i is di icul o a oid mic oplas ics in inal sea sal p oduc s wi hou u he
pu i ica ion s eps because seawa e con ains mic oplas ics[1-141-171]. In he las decade, esea che s ha e iden i ied
he p esence o mic oplas ics in ish and shell ish cap u ed in he wild and ob ained om aquacul u e a ms o ma ke s.
Mic oplas ics we e also isola ed om a ious p ocessed oods[1-141-172]. They we e in es iga ed in liquids such as
bee , honey and milk [1-141-168, 172]. The high concen a ions o mic oplas ics in bee samples equi ed u he
con i ma ion because s aining and isual coun ing may ha e o e es ima ed he numbe o pa icles [1-141-172].
Al hough suga con ains nea ly as much mic oplas ic as sea sal , he only s udy on mic oplas ics om suga s did no
use spec oscopic iden i ica ion me hods, and i migh include o he pa icles a he han mic oplas ics. Suga migh
also be con amina ed wi h mic oplas ic du ing p ocessing, equi ing u he in es iga ions [1-141-172]. D ied ood
such as land animal-based, Chinese adi ional medicine, p ocessed sea ood such as sa dines and sp a s, seaweed, d ied
ish, and eabags a e also con amina ed wi h mic oplas ics [1-141, 168, 169-172]. The high mic oplas ic concen a ions
in adi ional medicine is due o high mic oplas ic le els in he sou ce ma e ials [1-141-172]. Va ious s udies ha e
shown ha mic oplas ic causes ad e se oxici y, including e ec s on beha io al pa e ns, oxida i e s ess, hype
immune esponse, geno oxici y, ep oduc i e oxici y, de elopmen al oxici y, neu o oxici y, and gas oin es inal
oxici y [1-141-172].
3. Mic oplas ics: D inking wa e
Recen ly, i was announced ha he p esence o mic oplas ics in d inking wa e bo les is a opic o signi ican conce n
[1-141-172]. The p esence o mic oplas ic pa icles in d inking wa e aises conce ns abou hei po en ial impac on
human heal h. Tap wa e and bo led wa e ha e been iden i ied as signi ican pa hways o human exposu e o
mic oplas ics [9-141]. The p ima y sou ces o mic oplas ic in he en i onmen a e ex iles, medicine, and pe sonal ca e
i ems, while plas ic con aine s, ne s, ibe s, and iles cons i u e he seconda y sou ces [1-9-141-172]. Polye hylene (PE),
polys y ene (PS), polye hylene e eph hala es (PET), and polyp opylene (PP) cons i u e he mos abundan polyme s
in wa e [1-141]. These polyme s ind hei way in o wa e bodies om a ious packaging ma e ials such as plas ic
disposable bo les, bo le caps, ood packaging ma e ials, and plas ic bags which a e being disca ded [1-141]. E en
adi ionally pu e g oundwa e becomes ulne able o mic oplas ic in il a ion h ough ainwa e o plas ic deb is
leaching [7-9-141-172]. The use s age (e.g., opening he bo les) also con ibu es o mic oplas ics con amina ion
because physical s ess, hea , and ab asion can elease mic oplas ics in o he wa e . Packaging ma e ials, such as bo le
caps and polye hylene oils, can also elease mic oplas ics [7-9-141-172].The oxici y o mic oplas ics is known o
inc ease wi h dec easing size o he pa icle, as smalle pa icles ha e highe chances o pene a ing deepe in o he
o gans [1-141-172]. Mic oplas ics can en e he human body h ough inges ion, inhala ion, and de mal abso p ion
leading o hei accumula ion in a ious o gans and issues. These isks include he p esence o oxic chemicals,
accumula ion wi hin he body, p omo ion o mic obial g ow h, and ini ia ion o in lamma ion, due o unique
cha ac e is ics such as hyd ophobici y [1-141-172]. Mic oplas ics in bo led wa e esul om plas ic deg ada ion
du ing p oduc ion, anspo a ion, and use [9-141-172]. Besides hei own e ec s, mic oplas ics in d inking wa e pose
heal h isks by leaching addi i es/chemicals ha can cause damage o he human diges i e, neu oendoc ine,
ep oduc ion, and o he sys ems [9-141]. These chemicals can en e he human body h ough inges ion and de mal
con ac [9-141-172]. Imp ope ly disposing o disposable plas ic p oduc s, despi e hei con enience, aises signi ican
conce ns due o he ad e se impac s on he en i onmen and human heal h [7-9-172]. The accele a ed deg ada ion
poses signi ican isks, as he eleased o ganics a e iden i ied as e iological, ca cinogenic, and mu agenic, con ibu ing
o a ious ha m ul e ec s on he body [7-9-172]. These e ec s include oxida i e s ess, impai men s in he
gas oin es inal and ep oduc i e sys ems, me abolic dis u bances, and li e changes [7-9-172]. In India, he ecological
isk posed by mic oplas ics ound in coas al sedimen s has been e alua ed using a comp ehensi e analysis o me ada a,
employing h ee key indices: he Polyme Haza d Index (PHI), Pollu ion Load Index (PLI), and Po en ial Ecological Risk
Index (PERI) [10]. These indices p o ide a quan i a i e assessmen o sedimen quali y and he associa ed h ea le els
[10-141-172].
4. Mic oplas ic : De ec ion Me hods
A numbe o luo escen dyes, including Nile ed, Rhodamine B, Sa anin T, and luo escein iso phospha e, can label
plas ic polyme s and hence a e used in he de ec ion o mic oplas ics [1-141-172]. Among hese, Nile ed has been used
widely as a apid me hod o de ec ing mic oplas ics [1-141-166-172]. Fu he mo e, Fou ie ans o m in a ed (FTIR)
o Raman spec oscopy me hods a e commonly used o he de ec ion o oxic polyme chemicals [1-141-172]. Mo e
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2809-2824
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ad anced me hods, such as mic o FTIR, py olysis-GC/MS, and he mo g a ime ic analyse -GC/MS echniques, ha e
also been used o iden i y mic oplas ics wi h high p ecision. In addi ion o his, a numbe o animal models ha e been
used o e alua e he oxici y o mic oplas ic polyme s [1-141-166-172]. The li e a u e epo s he use o in e eb a es
like Caeno habdi is elegans, Daphnia magna, and A emia, and e eb a es such as mice, a s, oden s, and zeb a ish [7-
9-141-172]. These mic oplas ics ha e he po en ial o al e ma ine ecosys ems and pose heal h isks o aqua ic species
and, in u n, o humans [7-9-141-166-172].
5. Sepe a ion o Mic oplas ics om Sample o In e es
On he basis o li e a u e su ey, mic oplas ics a e usually collec ed om he samples by sie ing o dissec ion om
animal issue o in e es . Nex , mos samples will be ea ed wi h eagen s o chemically diges o dissol e he ma ix
such as KOH, HNO3, o H2O2 be o e ying o sepa a e he mic oplas ics by il a ion, o g a ime ically by u ilizing
di e ences in densi y wi h o wi hou cen i uga ion [1-141-166-172]. Fo una ely, he chemical s abili y o he
mic oplas ic pa icles o en allows analys s o dissol e and diges a ious ma ices while lea ing he polyme ic pa icle
in ac [1-141-166-172]. Howe e , ca e mus be aken—especially when using ni ic acid. One o he common me hods
used o diges samples is he use o a solu ion o concen a ed (30–35%) hyd ogen pe oxide (H2O2) o emo e o ganic
ma e [1-141-172]. In one o he s udy, diges ed esidues o wa e samples we e mixed wi h 20 mL o 30% hyd ogen
pe oxide a 60o Celsius o 72 h o diges o ganic ma e ial, hen he polyme pa icles in he diges ed samples we e
sepa a ed by densi y o e nigh using sa u a ed sodium chlo ide solu ion (D = 1.2 g/mL) [1-141-172]. In ano he s udy,
mic oplas ics p esen in ai samples we e collec ed and he sample il e s we e washed be o e diges ion in 35 mL o
30% H2O2 a oom empe a u e o 10 days o elimina e o ganic ma e ials [1-141-166-172]. The emaining H2O2
solu ion was hen acuum- il e ed h ough il e pape wi h a 2-mic on po e size o emo e any emaining pa icle
ma e [1-141-172]. Nex , 50 mL o a sa u a ed ZnCl2 solu ion wi h a densi y o 1.6 o 1.8 g/cm3 was added o each il e ,
and he mix u e was agi a ed o i e minu es a 350 pm [1-141-172]. Then, he samples we e allowed o emain s ill
o one ull hou be o e being cen i uged o 3 min a 4000 pm o collec all mic oplas ics [1-141-166]. The p e ious
li e a u e sugges s ha be o e he chemical analysis o mic oplas ics, samples mus be chemically diges ed, and he
mos common solu ion used o his pu pose is he 30% H2O2 [1-141-172]. Sepa a ion by densi y ( loa ing) o
mic oplas ic pa icles is a p ac ical and common s ep o easily pu i y mic oplas ics p io o chemical analysis [1-141-
172]. In gene al, he p o ocol o sampling, il a ion, chemical diges ion, and densi y sepa a ion is ou inely used o he
sample p epa a ion s eps o mic oplas ics analysis [1-141-172].
6. Physical and Chemical Cha ace e iza ion o Mic oplas ics
A e sepa a ing mic oplas ics om he sample o in e es , he nex s ep is physical and chemical cha ac e iza ion [1-
141-166-172]. Visual inspec ion is he quickes and he mos popula way o iden i y suspec ed mic oplas ic pa icles
[142-172]. Wi h he aid o op ical mic oscopes, mic oplas ic pa icles’ size, shape, and colo can be cha ac e ized [1-
141-172]. This me hod has he ad an age o being he simples , lowes cos , and allowing o he la ges di e si y o
mic oplas ic o be de ec ed in e ms o size (p ima y diame e o leng h), colo , and o m (as ibe , ilm, agmen , and
sphe ule). Dyes p oduce di e en colo s which allow us o isually de ec he ype, shape, and size o mic oplas ic [1-
141-172].
Mul iple dyes used include Oil Red EGN, Eosin B, Rose Bengal, Hos asol Yellow 3G, and NR (Nile Red) we e used in he
de ec ion o mic oplas ics [1-141-172]. In es iga o s soaked pa icles in he dyes o di e en du a ions be ween 5 min
and 66 h. Nile Red was chosen as he op imal s ain since i has he highes le els o adso p ion and luo escence in ensi y
[1-141-172]. When exposed o blue ligh , he dye will luo esce and simple pho og aphy wi h an o ange il e is used o
ind luo escence emission. Fluo escen pa icles can be ecognized and coun ed using image analysis [1-141-172].
Pa icles ha we e as small as a ew mic ome e s can be de ec ed using magni ied images ha can be eco ded and
iled/a anged o co e he en i e il e a ea. In e es ingly, Nile Red’s sol a och omic p ope ies p o ide he
oppo uni y o plas ic ca ego iza ion based on he su ace pola i y ai s o iden i ied pa icles [1-141-172]. I was
es ablished ha an incuba ion pe iod o be ween 5 min and 66 h and a dye concen a ion o be ween 1 and 1000 g mL-
1 we e ideal o isibili y. A wo king concen a ion o 10 g m-1 p oduced a nice balance be ween backg ound signal,
isibili y, and speed [1-141-172]. An addi ional echnique commonly used o cha ac e ize he size, shape, and chemical
composi ion o mic oplas ics is Scanning Elec on Mic oscopy (SEM). This echnique is used o image and measu e
objec s wi h di e en diame e s anging om millime e s o nanome e s in size [1-141-172]. The SEM echnique is
capable o nanome e spa ial esolu ion. An e ec i e and widely used me hod o iden i ying mic oplas ics is FTIR
mic ospec oscopy [1-141-172]. The signal is dependen on a change in molecula dipole momen occu ing du ing a
molecula ib a ion [1-141-166]. Ano he powe ul me hod o analysis o mic oplas ic pa icles is Raman
spec oscopy. Raman spec oscopy is ano he o m o ib a ional spec oscopy; howe e , in con as o FTIR, Raman
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2809-2824
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mic oscopy is mo e app op ia e o small mic oplas ics less han 20 µm [1-141-172]. Raman spec oscopy is based on
he Raman e ec , whe eby he equency o a small po ion o dispe sed o sca e ed adia ion emana ing om a sample
di e s om he equency o monoch oma ic inciden ligh . On he basis o li e a u e su ey, i is clea ha no pe ec
echnique exis s o mic oplas ic analysis which can p o ide bo h comp ehensi e chemical iden i ica ion and high-
esolu ion imaging capabili y [1-141-172].
7. Mic oplas ic De ec ion Me hods
Following a e he ew me hods used o he de ec ion o mic oplas ics in plas ic wa e bo les and ood ma e ial.
• Acco ding o he me hod desc ibed by Mohan e al., (2023) [1], 20 di e en b ands o plas ic wa e bo les (500
mL capaci y) we e pu chased in iplica es om local s o es in Mangalu u (n = 60), Ka na aka S a e, India[1].
A luo escence s aining me hod u ilizing Nile Red (NR), a lipophilic dye, was used o isualize he mic oplas ics
[1-3-172]. Nile Red (NR), can de ec mic oplas ics quickly and iden i y he polyme ic na u e o a pa icle
wi hou he need o ex a spec oscopic in es iga ion [1-3-172]. The wa e samples om each bo le we e
pou ed in o a s oppe ed glass bo le (1000 mL) unde he lamina hood and we e incuba ed wi h Nile ed (NR)
solu ion (10 μg/mL) o 30 min in he da k [1-3]. Nile Red solu ion (Sigma Ald ich) was p epa ed in ace one a
a concen a ion o 1 mg/mL o yield a wo king solu ion o 10 μg/mL and was s o ed a 4◦C in an ambe bo le
[1-4]. A e incuba ion, he samples we e acuum il e ed ia a glass mic o ibe il e pape [1-4]. Acco ding o
he me hod desc ibed by Mohan e al., (2023) [1], a e il a ion, he il e pape s we e kep in a s e ile glass
Pe i pla e and we e obse ed unde a luo escence mic oscope [1-4]. The il e pape was ma ked and di ided
in o quad an s o easy coun ing and so ing o he pa icles while imaging and cha ac e izing he shapes [1-4].
Mohan e al., (2023) [1] epo ed ha RO- ea ed wa e samples s o ed in glass con aine s we e used as a
nega i e con ol [1-4]. In addi ion, wa e samples ob ained om he plas ic bo les bu no incuba ed wi h Nile
Red we e p ocessed as desc ibed abo e and we e imaged o de e mine he backg ound luo escence [1-4-130].
To cha ac e ize he ype o polyme , 100 mL o he samples was il e ed wi hou Nile Red (NR) ea men [1-
4]. The il a e was hen washed wi h 200 μl o deionized wa e , collec ed in a glass ube, and subjec ed o
Fou ie - ans o m in a ed spec oscopy (FTIR) analysis [1-6]. This s udy by Mohan e al., (2023) [1], p o ides
e idence o he concep ha mic oplas ics a e abundan in he en i onmen , hey can en e he body h ough
wa e , can accumula e in a ious o gans, and can igge oxida i e s ess [1-172]. I would be in e es ing o
obse e he changes ha happen du ing long- e m exposu e o mic oplas ics and he esul ing changes in he
physiological and biochemical p ocesses in a cell [1-172]. Acco ding o he me hod desc ibed by Mohan e al.,
(2023) [1], Zeb a ish emb yos exposed o di e en concen a ions o luo escen - agged polye hylene
mic oplas ics (PE-MPs) (10–150 μm) showed accumula ion pa e ns a di e en ime poin s in a ious o gans
[1]. This s udy by Mohan e al., (2023) [1], also con i med ha mic oplas ics o a ious ypes we e de ec ed in
di e en b ands o packaged d inking wa e a ailable in India [1]. Nile Red s aining can be a simple and
e ec i e me hod o he de ec ion o plas ic polyme s [1-172]. FTIR esul s indica ed he abundance o
polye hylene, polys y ene, and polyamide in he samples [1]. Polye hylene polyme s al e ed he edox balance
in zeb a ish [1]. Co ela ions be ween oxida i e s ess and DNA damage we e obse ed upon mic oplas ic
exposu e o zeb a ish [1].
• Al hough washing wi h deionized wa e and hen isual inspec ion wi h o wi hou s aining is con enien o
clean ma ices, alse-posi i e de ec ion o mic oplas ics is challenging o a oid [1-141-172].
• The simples app oach o physical cha ac e iza ion is isual iden i ica ion, simply by naked eye obse a ion
using a dissec ing o s e eomic oscope [1-172]. This me hod is limi ed o mic oplas ic sizes in he ange o 2–
5 mm and is o en p one o e o when iden i ying mic oplas ics om complex en i onmen al samples. can be
used o coun he numbe o mic oplas ics in a sample and conduc a c ude mo phological s udy [1-172].
Howe e , i is limi ed by a measu emen e o , ha is he unc ion o pa icle size, i.e., he smalle he size, he
highe he pe cen age e o [1-141-172]. Wi h he use o a ligh mic oscope, la ge pa icles be ween 1 and 5
mm can be iden i ied. The ligh allows o he assessmen o colo , shape and helps dis inguish plas ic and non-
plas ic pa icles [1-166]. Elec on mic oscopes can also be used o isualize he size, shape, colou , and ex u e
o MPs wi hin he size ange o 0.5 o 5 mm [1-166-172]. Ad anced equipmen like came as in e aced wi h he
mic oscope and image e ining so wa e may be u ilized o de ec smalle mic oplas ics and imp o ed image
esolu ion o clea ly see de ails in he su ace mo phology [1-166-172]. Fu he mo e, o iden i y mic oplas ics
in he aqua ic en i onmen , luo escen o lipophilic dyes p o e use ul as hey can help easily isualize s ained
mic oplas ics unde a mic oscope [1-166-172].
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• Ano he popula oxida ion me hod is he use o Fen on’s eagen . This me hod is sugges ed by he Na ional
Oceanic and A mosphe ic Adminis a ion, USA, o ma ine o ganisms, al hough he me hod needs o be es ed
o a di e si y o o ganic ma ices [1-141-172].
• Diges ion wi h alkaline solu ions such as KOH and NaOH ha e p edominan ly been used o diges ing ish and
shell ish. I is ad an ageous o des oying p o eins and o he so issues. Sui able ex ac ion eco e y was
ound o polye hylene e eph hala e (PET) and high-densi y polye hylene [1-141-172]. Howe e , pH-sensi i e
polyme s such as nylon and polyes e can be dis up ed a high pH [1-141-172]. Va ious s ong acid solu ions
(e.g., HNO3, HCl/HNO3, and HClO4) ha e been used o diges he so issues o ish, mussels, and o he o ganisms
[141,142]. Simila o s ong basic solu ions, he issues we e success ully decomposed, al hough low pH also
led o he decomposi ion o pH-sensi i e polyme s [1-172].
• Mic oscopy, including op ical and elec on mic oscopy, is a s anda d me hod o iden i ying mic oplas ic based
on hei size and shape due o i s simplici y and low cos [142-166-172]. Howe e , his app oach is limi ed o
mic oplas ic, is ime-in ensi e and elies hea ily on he analys ’s judgmen , making i suscep ible o e o s
in luenced by en i onmen al ac o s and sample impu i ies. Fluo escence s aining is now widely applied as a
supplemen a y echnique o enhance mic oplas ic iden i ica ion by mic oscopy [1-142-166-172]. This me hod
in ol es s aining o mic oplas ic wi h hyd ophobic dyes and using speci ic wa eleng hs o igge luo escence,
which aids in de ec ion unde specialized mic oscopes [1-142-172]. Howe e , issues like alse posi i es due o
s aining o o ganic ma e ials o in e e ence om na u al luo escence in samples emain challenges. The
de elopmen o new luo escen dyes ha can ecognize speci ic mic oplas ic would g ea ly ad ance he ield
by imp o ing he accu acy o mic oplas ic iden i ica ion in complex en i onmen al samples. Ad anced me hods
like scanning elec on mic oscopy (SEM) and a omic o ce mic oscopy (AFM) a e inc easingly being used o
s udy he smalle dimensions o nanoplas ics [1-142-172]. SEM p o ides high- esolu ion imaging o
mo phological ea u es, especially su ace cha ac e is ics. When pai ed wi h ene gy-dispe si e X- ay
spec oscopy, i can also p o ide aluable insigh s in o chemical composi ion [142-172]. Howe e , SEM is slow
and equi es ime-in ensi e sample p epa a ion. In con as , AFM is a p omising ool o mic o- and nanoscale
analysis due o i s abili y o cap u e high- esolu ion images di ec ly om he sample wi hou p e ea men [1-
142-171]. Fu he mo e, AFM is as , simple and can dis inguish ma e ial ypes wi hin polyme blends, de ec ing
compounds like hea y me als adso bed on o mic oplas ic su aces [1-142-166].
• The wo p edominan me hods used o mic oplas ic iden i ica ion in ood we e isual inspec ion unde
dissec ion mic oscope wi h o wi hou s aining and he abso p ion o e lec ion o IR wi h FT-IR o Raman
spec oscopy [1-141-172].
• Two cu en app oaches—coun ing mic oplas ics wi h mic oscopy and des uc i e mic oplas ic de ec ion wi h
he mal analysis—can be complemen a y [1-172]. In addi ion, con amina ion and decon amina ion o
mic oplas ics du ing ood p ocessing and cooking a e impo an as mic oplas ic exposu e o people is p ima ily
om he consumed inal p oduc s, no on hei ing edien s [1-142-166].
• The h ee main me hods o de ec ing and quan i ying mic oplas ic concen a ions in wa e a e FTIR
Spec oscopy, py-GC/MS, and Raman Spec oscopy [1-141-172]. FTIR and Raman can de e mine he numbe
o mic oplas ic pa icles by plas ic ype and size ange, whe eas py-GC/MS can quan i y concen a ions o
speci ic ypes o mic oplas ics in mg/l [1-142-172]. These me hods can also be used o de ec mic oplas ics in
sludge and soil samples. Raman spec oscopy plays a key ole in iden i ying he ypes and o igins o
mic oplas ics. I is a pa o he e o s o de elop policies and p ocedu es o con olling he amoun o
mic oplas ics in oduced in o ecosys em [1-141-172].
• Mic o- and nanoplas ics a e in ou ood, wa e and he ai we b ea he. They a e showing up in ou bodies,
om es icles o b ain ma e [12, 13-141-172]. Nano and mic oplas ics a e byp oduc s o deg ading plas ic
ma e ials such as lunchboxes, cups and u ensils. As e y small pa icles wi h a la ge su ace a ea, nanoplas ics
a e pa icula ly conce ning o human heal h due o hei inc eased abili y o abso b oxins and pene a e
biological ba ie s wi hin he human body. De ec ing hese plas ics ypically equi es skilled pe sonnel and
expensi e equipmen [12, 13-172]. Now, UBC, Vancou e , Canada esea che s ha e de eloped a low-cos ,
po able ool o accu a ely measu e plas ic eleased om e e yday sou ces like disposable cups and wa e
bo les [13-172]. The de ice, pai ed wi h an app, uses luo escen labeling o de ec plas ic pa icles anging
om 50 nanome es o 10 mic ons in size – oo small o be de ec ed by he naked eye – and deli e s esul s in
minu es. They c ea ed a small, biodeg adable, 3D-p in ed box con aining a wi eless digi al mic oscope, g een
LED ligh and an exci a ion il e [13]. To measu e he plas ics, hey cus omized MATLAB so wa e wi h
machine-lea ning algo i hms and combined i wi h image cap u e so wa e [13]. The esul is a po able ool
ha wo ks wi h a sma phone o o he mobile de ice o e eal he numbe o plas ic pa icles in a sample. The
ool only needs a iny liquid sample – less han a d op o wa e – and makes he plas ic pa icles glow unde he
g een LED ligh in he mic oscope o isualize and measu e hem [13, 149]. The esul s a e easy o unde s and,
whe he by a echnician in a ood p ocessing lab o jus someone cu ious abou hei mo ning cup o co ee [13].
The ool is cu en ly calib a ed o measu e polys y ene, bu he machine-lea ning algo i hm could be weaked
Wo ld Jou nal o Ad anced Resea ch and Re iews, 2025, 26(02), 2809-2824
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o measu e di e en ypes o plas ics like polye hylene o polyp opylene [13]. Nex , he esea che s aimed o
comme cialize he de ice o analyze plas ic pa icles o o he eal-wo ld applica ions [13]. To educe plas ic
inges ion, i is impo an o conside a oiding pe oleum-based plas ic p oduc s by op ing o al e na i es like
glass o s ainless s eel o ood con aine s [12, 13]. The de elopmen o biodeg adable packaging ma e ials is
also impo an o eplacing adi ional plas ics and mo ing owa ds a mo e sus ainable wo ld.
• The global mic oplas ic de ec ion ma ke is expe iencing signi ican g ow h due o ising en i onmen al
conce ns, egula o y p essu es, and ad ancemen s in de ec ion echnology [1-143-172]. Wi h mic oplas ic
con amina ion ound in wa e , soil, ai , and ood p oduc s, indus ies a e in es ing in high-p ecision
spec oscopy, ch oma og aphy, and mic oscopy-based solu ions o ensu e accu a e de ec ion and compliance
[1-143-172]. Key playe s like The mo Fishe Scien i ic, Agilen Technologies, B uke , and Pe kinElme a e
leading he inno a ion in eal- ime moni o ing and high-sensi i i y es ing me hods [1-143].
• To o e come he challenges o ime and labo -in ensi e, mic oplas ic de ec ion echniques, esea che s a e
inc easingly adop ing machine lea ning and au oma ion. These echnologies can p ocess la ge da ase s wi h
g ea e speed and accu acy, aining algo i hms o de ec mic oplas ic mo e e icien ly [142-166-172]. Machine
lea ning models can analyze spec al da a o images, iden i ying pa e ns ha dis inguish mic oplas ic om
o he pa icles [130-142-172]. Se e al s udies ha e now p o en he po en ial o machine lea ning o
mic oplas ics iden i ica ion using SEM, luo escence, Ramen spec oscopy and FTIR. One s udy in oduced
Plas icNe , a deep lea ning model speci ically ained o ecognize mic oplas ics om images gene a ed by FPA-
based mic o-FTIR spec oscopy [130-142-166-172]. Plas icNe , ained on spec a om 11 ypes o i gin
plas ic pa icles, achie ed o e 95% classi ica ion accu acy, demons a ing he huge po en ial o deep lea ning
in mic oplas ic de ec ion [120-142-172]. Howe e , while he model shows high accu acy o i gin plas ics, his
model's e icacy on en i onmen ally sou ced mic oplas ic emains un es ed [142-166]. In ano he s udy, a
da ase o o e 64,000 Raman spec a om 47 en i onmen al o was ewa e samples was used o de elop a
human-compu e hyb id app oach. This me hod achie ed high ecall (≥99.4%) and p ecision (≥97.1%) o
iden i ying mic oplas ic and educed he anno a ion ime om hou s o unde one hou pe sample compa ed
o human-only analysis [120-142-172].
• Cu en ly, in si u de ec ion and quan i ica ion o mic oplas ic is di icul o e en impossible, because o a lack
o applicable me hods. Ce ain en i onmen s, like was ewa e ea men plan s and wa e -in ensi e indus ies,
con ain high le els o o ganic and ino ganic solids, complica ing he de ec ion o low-abundance mic oplas ic
wi hou sample p e ea men [142-172]. En i onmen al ac o s, such as empe a u e and p essu e luc ua ions
in na u al wa e bodies, add u he complexi y by al e ing condi ions a di e en dep hs, a ec ing he
p ope ies o mic oplas ic locally. Recen ad ancemen s in compac ligh sou ces, de ec o s and op ical
componen s ha e led o he a ailabili y o po able and handheld pho ome e s and spec ome e s o
en i onmen al moni o ing [142-172]. Among hese, comme cial po able de ices based on luo escence, FTIR
and Raman ha e shown p omise. Fo example, a cos -e ec i e po able Raman senso has been designed o
de ec mic ome e -sized magne ic plas ic pa icles in wa e using a qua z cu e e [142-172]. Addi ionally, a
ecen s udy u ilized a po able pho ome e and luo escen s aining o measu e he p esence o mic oplas ic
in wa e samples, ep esen ing signi ican p og ess owa d accessible, ield- eady de ec ion me hods [142-
172].
• Mic owa e echnology is a no el me hod o moni o ing mic oplas ics. I is a e y p omising app oach o
quickly de e mining he size and concen a ion o selec i e mic oplas ic sensing [169]. Al hough he e a e some
limi a ions, and a e wo king on ways o o e come he weaknesses by enhancing he mic owa e pa o he
de ice and imp o ing he sensi i i y o he senso [169]. This enhancemen migh allow us o de ec
mic oplas ic pa icles as accu a ely as op ical spec oscopic me hods bu would be much as e and cheape
[140-169]. Mic owa es a e highly sensi i e o he p ope ies o he ma e ials wi h which hey in e ac .
Mic owa e senso s a e based on he di e ence in pe mi i i y o he hos medium (e.g., wa e ) and he
mic oplas ic con aminan [169]. This con as is high, enabling esea che s o accu a ely coun he numbe o
pa icles p esen , e en a low concen a ion le els [169]. Mic owa e senso s also can be combined wi h plana
echnology o ob ain compac , ligh , obus , and low-p ice ab ica ion[169]. The e a e s eng hs and
weaknesses associa ed wi h any de ec ion me hodology, including mic owa e sensing. One downside is ha
mic owa e senso s canno de ec pa icles smalle han 20 mic ome e s (in samples wi h concen a ions less
han 1000k pa icles/L o hose wi h concen a ions less han 50k pa icles/L) [140-169]. Ano he challenge
wi h his me hod is ha i canno di e en ia e be ween wo di e en plas ics in a sample, bu his could be
add essed in he u u e as he echnology imp o es [169].
Deb aj and Mulky (2025) [168] a e o he opinion ha cu en ly, he mos commonly used me hod o mic oplas ic
iden i ica ion is he use o a s e eomic oscope [168]. I has a high e o a e and o en esul s in alse posi i es.
Spec oscopy me hods a e mo e e icien . Howe e , hey equi e expensi e and bulky equipmen and ained pe sonnel
o ope a ion [168]. Mo eo e , depending on he en i onmen al sample, hey equi e long p e- ea men s eps. Mul iple
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da ase s gene a ed could be so ed and classi ied wi h he help o machine lea ning ools [168]. Howe e , associa ed
issues include high dimensionali y, limi ed accu acy, low pe o mance, and edundancy [166-169]. Con en ional
me hods o mic oplas ic so ing ely g ea ly on he physical s a e o he sample, he p o ocol o sample p epa a ion,
and he possibili y o con aminan s. Fu he mo e, new echniques con inue o de elop, om mechanisms ha exploi
he elec os a ic p ope ies o MPs, o mic o luidics and e en he use o he In e ne o Things [168].
While no single echnique p o ides a one-size- i s-all solu ion o de ec ing mic oplas ic ad ancemen s in spec oscopy,
mass spec ome y, imaging and AI-d i en app oaches e apidly imp o ing ou abili y o ack hese pollu an s [142-
166-169]. The challenge now is o make hese echnologies as e , mo e cos -e ec i e and scalable o widesp ead
en i onmen al moni o ing. By de eloping po able ools and in eg a ing machine lea ning, we may soon ha e he
capabili y o map he global dis ibu ion o mic oplas ic, as well as dis inguish be ween di e en ypes o plas ics, ace
hei o igins and moni o hei mo emen h ough ecosys ems and ood chains [142-166-169].By enhancing ou
de ec ion capabili ies, he scien i ic communi y can wo k owa d in o med, e idence-based policies and in e en ions
aimed a educing plas ic pollu ion and p o ec ing bo h en i onmen al and human heal h [142-166-169].
Cu en ly, he de ec ion o mic oplas ics p ima ily elies on isual analysis using s e eoscopes o a combina ion o
spec oscopic echniques, including Fou ie ans o m in a ed spec oscopy (FT-IR) and Raman spec oscopy,
luo escen s aining me hods (e.g. Nile Red), he mal analysis, and ch oma og aphic analysis [1-96–98-172]. Howe e ,
hese me hods ha e limi a ions and d awbacks. Fo ins ance, he use o Nile Red luo escen s aining may complica e
he iden i ica ion o speci ic polyme ic ma e ials [1-99-172]. Some me hods a e subjec i e, ime-consuming, and p one
o e o s due o he a ying appea ances and p ope ies o mic opls ics. The e o e, mo e objec i e c i e ia and he
assis ance o AI o da a analysis a e needed o help scien is s accu a ely iden i y he mo phological ea u es o
mic oplas ics. The e is an u gen need o scien is s o de elop mo e ime-e icien and p ecise me hods o de ec ing
MPs in he en i onmen and human bodies, o add ess he challenges aced in mic oplas ics de ec ion. Wi h
ad ancemen s in mic oplas ics de ec ion echnology, especially in he explo a ion o me hods o de ec ing
mic oplas ics wi hin he human body, i is an icipa ed ha mo e di ec e idence o he co ela ion be ween
mic oplas ics and human heal h will be unco e ed in he u u e.
8. Conclusion
Mic oplas ics ha e been ound in a ious en i onmen al media, including soil, wa e , ai , and ha e been shown o ha e
nega i e impac s on wildli e. Mic oplas ics ha e en e ed d inking wa e ia a ious pa hways, aising conce ns abou
hei po en ial heal h impac s. Mul iple dyes used include Oil Red EGN, Eosin B, Rose Bengal, Hos asol Yellow 3G, and
NR (Nile Red) we e used in he de ec ion o mic oplas ics. When exposed o blue ligh , he dye will luo esce and simple
pho og aphy wi h an o ange il e is used o ind luo escence emission. Fluo escen pa icles can be ecognized and
coun ed using image analysis. The h ee main me hods o de ec ing and quan i ying mic oplas ic concen a ions in
wa e a e FTIR Spec oscopy, py-GC/MS, and Raman Spec oscopy. FTIR and Raman can de e mine he numbe o
mic oplas ic pa icles by plas ic ype and size ange, whe eas py-GC/MS can quan i y concen a ions o speci ic ypes o
mic oplas ics in mg/l. These me hods can also be used o de ec mic oplas ics in sludge and soil samples. Raman
spec oscopy plays a key ole in iden i ying he ypes and o igins o mic oplas ics. I is a pa o he e o s o de elop
policies and p ocedu es o con olling he amoun o mic oplas ics in oduced in o ecosys em. The e a e se e al, well-
es ablished me hods o de ec ing and moni o ing mic oplas ic pa icles – he mos common being Fou ie - ans o m
in a ed (FTIR) and Raman spec oscopy. Al hough hese echniques a e e y accu a e and can de ec he ypes o
plas ic in he sample, hey equi e in ense aining o he echnician and bulky and expensi e measu emen equipmen .
They also use o line me hods and equi e samples di ec om wa e acili ies o lab expe imen s. O e all, hese
app oaches end up being ime-consuming, labo -in ensi e, and expensi e o esea che s, especially when hey ha e a
la ge numbe o samples o p ocess, The mos signi ican challenge in mic oplas ic de ec ion, much like mic oplas ic
ex ac ion and sampling, is he lack o a s anda dized me hod o iden i ica ion. Due o he ple ho a o polyme
composi ions and nume ous complex en i onmen al ma ices, i is almos impossible o ha e a singula iden i ica ion
p o ocol. Mo eo e , i is also dependen on he a ailabili y and accessibili y o he ins umen s, echnicians, and ela ed
ma e ials. Di e en ins umen s p o ide di e en physico-chemical p ope y in o ma ion, and each has i s own se o
ad an ages and limi a ions.
Compliance wi h e hical s anda ds
Disclosu e o con lic o in e es
No con lic o in e es o be disclosed.
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