1
Mul i esidue analy ical me hod o pha maceu icals and pe sonal ca e p oduc s in 1
sewage and sewage sludge by online di ec imme sion SPME on- ibe de i a iza ion – 2
GCMS 3
4
Rebeca López-Se na () • Da id Ma ín-de-Jesús • Rubén I us a-Ma a • Ped o An onio 5
Ga cía-Encina • Raquel Leb e o • Ma ía Fdez-Polanco • Raúl Muñoz 6
7
Depa men o Chemical Enginee ing and En i onmen al Technology, Uni e si y o 8
Valladolid, C/ D . Me gelina, s/n, 47011 Valladolid, Spain 9
10
() Co esponding au ho : 11
Rebeca López-Se na 12
e-mail: ebeca.lopezs[email p o ec ed]a.es 13
14
Abs ac 15
The wo k he e p esen ed aimed a de eloping an analy ical me hod o he 16
simul aneous de e mina ion o 22 pha maceu icals and pe sonal ca e p oduc s, 17
including 3 ans o ma ion p oduc s, in sewage and sludge. A me iculous me hod 18
op imiza ion, in ol ing an expe imen al design, was ca ied ou . The de eloped 19
me hod was ully au oma ed and consis ed o he online ex ac ion o 17 mL o wa e 20
sample by Di ec Imme sion Solid Phase Mic oEx ac ion ollowed by On- ibe 21
De i a iza ion coupled o Gas Ch oma og aphy - Mass Spec ome y (DI-SPME – On-22
ibe De i a iza ion – GC - MS). This me hodology was alida ed o 12 o he ini ial 23
compounds as a eliable ( ela i e eco e ies abo e 90% o sewage and 70% o 24
sludge; epea abili y as %RSD below 10% in all cases), sensi i e (LODs below 20 ng L-1 25
in sewage and 10 ng g-1 in sludge), e sa ile (sewage and sewage-sludge samples up o 26
15,000 ng L-1 and 900 ng g-1, espec i ely) and g een analy ical al e na i e o many 27
medium- ech ou ine labo a o ies a ound he wo ld o keep up wi h bo h cu en and 28
o ecas en i onmen al egula ions equi emen s. The emaining 10 analy es ini ially 29
conside ed showed insu icien sui abili y o be included in he inal me hod. The 30
me hodology was success ully applied o eal samples gene a ed in a pilo scale 31
sewage ea men eac o . 32
2
33
Keywo ds: DI-SPME • GC-MS • On-Fibe De i a iza ion • PPCPs • Sewage sludge • 34
Was ewa e 35
36
1 In oduc ion 37
The de elopmen o analy ical me hodologies o he de e mina ion o 38
pha maceu icals and pe sonal ca e p oduc s (PPCPs) in en i onmen al ma ices has 39
boomed in he pas yea s. In his con ex , Zwiene and F immel [1] epo ed ha he 40
analysis o PPCPs has been adi ionally domina ed by Liquid Ch oma og aphy 41
de ec ed by andem Mass Spec ome ic (LC-MS/MS) echniques. Fische e al. [2] 42
ecen ly obse ed majo ends in he use o Ul a High Pe o mance Liquid 43
Ch oma og aphy (UHPLC) [3] and High Resolu ion Mass Spec ome y (HRMS) [4-6] like 44
Time O Fligh (TOF) and O bi ap [7] analyze s. Howe e , hese echniques equi e 45
cos ly ins umen a ion no a o dable by many labo a o ies wo ldwide. In con as , 46
Gas Ch oma og aphy coupled o single quad upole Mass Spec ome y (GC-MS) is an 47
analy ical con igu a ion a mo e common in ou ine analysis labo a o ies a ound he 48
wo ld, including de eloping coun ies. Despi e PPCPs a e mainly pola compounds and 49
no eadily analyzable by GC, Lopez-Se na e al. [8] ecen ly showed how GC-MS is a 50
alid ins umen al echnique o he analysis o eme ging con aminan s in 51
en i onmen al ma ices like sewage, when a de i a iza ion s ep is included in he 52
me hod. In e ms o sample p epa a ion, Solid-Phase Ex ac ion (SPE) ep esen s 53
nowadays he mos popula echnique o he ex ac ion o pollu an s om 54
en i onmen al aqueous samples, and ecen de elopmen s in his ield ha e mainly 55
ocused on SPE au oma ion [9]. In addi ion, a g ea e o has been la ely made o 56
de elop new analy ical me hodologies able o pe o m di ec analyses using 57
minia u ized equipmen , he eby achie ing high en ichmen ac o s, minimizing 58
sol en consump ion and educing was e [7, 10] in acco dance o he equi emen s o 59
g een analy ical chemis y. Solid-Phase Mic oEx ac ion (SPME) was i s ly de eloped 60
in he 1990s by Pawliszyn and cowo ke s [11]. Since hen many con igu a ions ha e 61
been success ully implemen ed, which can be classi ied in o s a ic and dynamic 62
echniques [12]. S a ic p ocedu es a e ypically ca ied ou in s i ed samples, including 63
ibe SPME, and cons i u e he mos common o ma o his echnique. Fibe SPME 64
3
u ilizes a so ben coa ing on he ou e su ace o a used silica ibe o ex ac he 65
analy e(s) om he sample ma ix in a p ocess ha occu s h ough di ec imme sion 66
(DI-SPME) o om he sample headspace in a closed con aine (HS-SPME) [10]. Thus, 67
analy es ha exhibi a high apo p essu e can be ex ac ed ei he by imme sing he 68
ibe in o he aqueous sample o by sampling i s headspace. In con as , analy es ha 69
exhibi a low apo p essu e could only be ex ac ed by imme sion. Fibe SPME has 70
become a e y popula echnique, especially o ola ile compounds, due o i s 71
simplici y, ela i ely sho ex ac ion ime, sol en - ee na u e, ull au oma ion 72
po en ial and easy coupling wi h ch oma og aphy [12]. These ad an ages e en ually 73
educe he con amina ion o he o iginal sample and he loss o analy es. In addi ion, 74
SPME can also be used o onsi e sample ex ac ion and is able o ob ain good esul s 75
e en o ace analy es in complex ma ices [12]. Howe e , i s applica ion o he 76
en i onmen al analysis o pola compounds has been poo ly explo ed, especially when 77
his sample p e ea men is coupled o GC. This applica ion implies he addi ion o a 78
de i a iza ion s ep, which is essen ial o he analysis o non- ola ile and/o 79
he molabile compounds by GC. Today, wo app oaches a e commonly used o ca y 80
ou de i a iza ion when SPME is he p e ea men echnique. The i s one, namely in-81
si u de i a iza ion, is based on he addi ion o he de i a izing agen di ec ly o he 82
sample and he collec ion o he de i ed ola ile analy es by SPME in he headspace o 83
a closed ial. In he second app oach, namely on- ibe de i a iza ion, analy e 84
ex ac ion occu s ia di ec ibe imme sion in he sample combined wi h a headspace 85
de i a iza ion by exposing he analy es-loaded ibe o he apo s o he de i a izing 86
agen . This second app oach is en i onmen ally and economically p e e ed, because 87
he de i a izing agen can be eused o a la ge numbe o analyses (wi h he 88
subsequen dec ease o eagen consump ion). 89
This s udy aimed a de eloping and op imizing a ully au oma ed me hod 90
consis ing o Online DI-SPME - On-Fibe De i a iza ion - GC-MS o he analysis o 19 91
PPCPs and 3 o hei T ans o ma ion P oduc s (TPs) in sewage (SW) and sludge (SS) 92
using s a is ical expe imen al design. To he au ho s’ knowledge, he e a e only wo 93
o he publica ions [13, 14] p oposing he use o his echnique o he analysis o 94
PPCPs in sewage and none o sludge. Howe e , none o hem included he le el o 95
4
au oma ion he e p esen ed. Finally, he analy ical limi a ions encoun e ed du ing he 96
applica ion o his inno a i e me hodology we e also discussed. 97
98
2 Ma e ial and me hods 99
2.1 Chemicals 100
The s anda ds o all PPCPs and hei TPs, p o ided in Table S1 as 101
supplemen a y da a, we e o high pu i y g ade (>95%). They we e pu chased om 102
Sigma-Ald ich (T es Can os, Mad id, Spain) as neu al non-sol a ed molecules, excep 103
o amoxicillin (acqui ed as ihyd a e), a o as a in (acqui ed as calcium sal ) and 104
diclo enac (acqui ed as sodium sal ). The iso opically labelled compounds Diclo enac-105
d4, Ibup o en-d3, Salicylic acid-d4, Nap oxen-d3, P opylpa aben-d7 and T iclosan-d3 106
we e ob ained om TRC Canada (To on o, ON, Canada). 107
Indi idual s ock solu ions a 1 g L-1 o bo h PPCPs s anda ds and iso opically-108
labelled-in e nal-s anda ds we e p epa ed on a weigh basis in me hanol (MeOH), 109
excep o he lu oquinolones (cip o loxacin, le o loxacin and no loxacin), which 110
we e dissol ed in a wa e -me hanol (H2O/MeOH) mix u e (1:1) con aining 0.2% / 111
hyd ochlo ic acid (HCl) due o hei low solubili y in pu e MeOH [15]. F om hem, a 112
s ock solu ion wi h all he analy es was hen p epa ed in MeOH a 20 mg L-1. Se ial 113
aqueous dilu ions we e subsequen ly p epa ed om i . A sepa a e mix u e o 114
iso opically labelled in e nal s anda ds and u he dilu ions we e also p epa ed. A e 115
p epa a ion, all s ock solu ions we e s o ed a -20 °C in da kness. 116
High pu i y sol en s, i.e., Sup aSol ® GC-MS g ade MeOH by Me ck Millipo e 117
(Mad id, Spain), LC-MS Ch omasol ® g ade E hyl Ace a e (EA) by Fluka (Mad id, Spain), 118
Sodium chlo ide (NaCl) and 37% HCl we e supplied by Pan eac (Ba celona, Spain). 119
Ace one, 99% pu e, was supplied by Co a cas (Bu gos, Spain). N- e -120
Bu yldime hylsilyl-N-me hyl i luo oace amide, wi h a pu i y >99%, (MTBSTFA), was 121
ob ained om Regis Technologies Inc. (Mo on G o e, IL, USA). SPME ibe s we e 122
pu chased om Supelco (T es Can os, Mad id, Spain). Milli-Q® g ade wa e was in-123
house p oduced. Helium 99.999% (He) was pu chased om Abelló Linde S.A. (Alcalá de 124
Hena es, Mad id, Spain). 125
5
126
2.2 Sewage analy ical me hodology 127
The de elopmen o he analy ical me hod, u he explained in Sec ions SD.1.1 128
and SD.1.2 wi hin he Supplemen a y da a (SD), was ca ied ou in Milli-Q® wa e and 129
alida ed o sewage as de ailed in Sec ion 3.2.1. In addi ion, he op imized me hod 130
based on Online DI-SPME – On-Fibe De i a iza ion – GC – MS was applied o he 131
analysis o aw and ea ed was ewa e om a pilo scale ac i a ed sludge eac o , and 132
he esul s a e p esen ed in Sec ion 3.2.2. 133
134
2.2.1 Online DI-SPME – On-Fibe De i a iza ion 135
Wa e samples (100 mL) we e supplemen ed wi h NaCl a 30 % (w ./ ol.). A e 136
s i ing o 20 min o assu e comple e dissolu ion, he esul ing wa e sample pH was 137
adjus ed o 3 by adding as ew d ops o dilu ed solu ions o HCl (1%, 0.1% and/o 138
0.01%) as needed. A olume o 17 mL o he esul ing solu ion was placed in a 20-mL 139
SPME ial along wi h 200 µL o an aqueous mix u e o he iso opically labelled in e nal 140
s anda ds a 0.5 mg L-1. 141
The esul ing ial was placed in he sample ack o a CTC PAL RSI au osample . A 142
SPME ool held a 2-cm long 50/30-µm hick 143
Di inylbenzene/Ca boxen/Polydime hylsiloxane (DVB/CAR/PDMS) S ableFlex/SS ibe 144
ha was p o ec ed inside a 23 Ga needle. The ully au oma ed DI-SPME me hod 145
included a ibe p e-condi ioning o 15 min a 270 °C in he spa e GC inle , ollowed by 146
120 min sample ex ac ion a a pene a ion dep h o 60 mm, which en ailed ha he 147
ibe was ully imme sed in he sample (DI-SPME). On- ibe de i a iza ion o he 148
analy es abso bed on o he ibe was hen ca ied ou by in oducing he ibe in 149
ano he 20-mL SPME ial con aining 1 mL o he de i a izing agen MTBSTFA o 48 150
min a a pene a ion dep h o 60 mm. Thus, he ibe was exposed o he apo s o he 151
MTBSTFA in he headspace o he ial. Bo h he DI-SPME and On-Fibe De i a iza ion 152
we e ca ied ou a a cons an empe a u e o 50 °C unde o bi al agi a ion a 500 pm 153
wi h a s i ing egime o 6s on / 30 s o . The ibe , loaded wi h he de i a ized 154
analy es, was hen aken o he GC inle connec ed o he GC column o deso p ion a 155
250 °C o 3 min. Finally, he ibe was pos -condi ioned o 15 min a 270 °C in he 156
spa e GC inle p io o he nex analysis. 157

6
158
2.2.2 GC – MS 159
Ch oma og aphic uns s a ed concomi an ly wi h ibe deso p ion in a pulsed 160
spli less mode a 250 °C in he spli /spli less back inle . A SPME injec ion slee e, 0.75 161
mm i.d., was used as a line . The es s we e pe o med in an Agilen 7890B GC Sys em 162
coupled o a 5977A MSD. A capilla y HP-5MS GC column (30 m leng h, 0.25 mm i.d., 163
0.25 μm ilm hickness) was used o he ch oma og aphic sepa a ion wi h He as 164
ca ie gas a a cons an low a e o 1.2 mL min-1. Injec o empe a u e was se a 250 165
°C, while he GC o en empe a u e inc eased om 70 °C (held o 3 min du ing ibe 166
deso p ion) o 120 °C a 20 °C min-1, hen o 250 °C a 10 °C min-1 and inally o 300 °C 167
(held o 5 min) a 5 °C min-1. The o al analysis ime o each GC un was 33.5 min. The 168
mul imode on GC inle was se a 270 °C in spli mode o acili a e he elimina ion o 169
esidual compounds du ing ibe p e- and pos -condi ioning. 170
Mass de ec ion was ob ained in elec on impac ioniza ion mode (70 eV) wi h 171
selec ed ion moni o ing (SIM) and a ilamen delay o 12 min. The GC–MS in e ace, 172
ion sou ce and quad upole empe a u es we e se a 280, 230 and 150 °C, 173
espec i ely. Quad upole esolu ion was se a low. Ta ge compounds we e eco ded 174
in i e acquisi ion windows along he un ime. Table 1 shows he p ima y (in i alics) 175
and he wo seconda y ions moni o ed pe compound. Acquisi ion s opped a min 26. 176
Ins umen con ol and da a acquisi ion we e pe o med by Agilen Technology Mass 177
Hun e B.07.03.2129 so wa e. 178
179
2.3 Sewage sludge analy ical me hodology 180
Ae obic sludge was used o de elop and alida e he me hodology u he 181
discussed in Sec ions SD.1.3 and 3.2.1, espec i ely. The sewage sludge analy ical 182
me hod was designed as ollows: 1) One hund ed millili e s o esh sludge sample 183
we e eeze-d ied. 2) A known amoun o d ied sludge (~800 mg) was weighed in o a 184
20-mL glass ial, along wi h 200 µL o a mix u e o he iso opically labelled in e nal 185
s anda ds a 20 mg L-1 in ace one. 3) The mix u e was ho oughly o ex-s i ed and 186
emained o e nigh o allow sol en e apo a ion and in e nal s anda d ixa ion. 4) A 187
olume o 12 mL o MilliQ® wa e a pH 9 was hen added o he ial, which was hen 188
igo ously o ex-s i ed o ob ain a homogenous suspension. 5) Then, he ial 189
7
unde wen Ul asound Assis ed Ex ac ion (UAE) o 30 min a oom empe a u e in a 190
JP Selec a Uni eba ul asonic ba h o 50 W and 60 Hz (Ba celona, Spain). 6) 191
Subsequen ly, he suspension was cen i uged o 5 min a 2,655 x g in a Fishe 192
Bioblock Scien i ic Cen i uge 2-16P (Mad id, Spain). 7) The esul ing supe na an was 193
hen collec ed wi h a glass pipe e and ans e ed o a 20-mL glass ial. 8) S eps 4-7 194
we e epea ed once mo e and he supe na an s we e pooled oge he . 9) The esul ing 195
solu ion was analyzed by Online DI-SPME – On- ibe de i a iza ion – GC-MS using he 196
op imized me hod desc ibed in Sec ion 2.2, excep o he addi ion o in e nal 197
s anda ds as hey we e al eady added in s ep 2. 198
199
2.4 Expe imen al design 200
As a i s app oach, a sc eening design was ca ied ou . Hence, he key 201
pa ame e s in luencing he pe o mance o he Online DI-SPME – On-Fibe 202
De i a iza ion me hodology we e iden i ied o he de elopmen o he ins umen al 203
leg o bo h sewage and sludge me hods. As a esul , a o al o 18 pa ame e s we e 204
so ed ou in ou ca ego ies, depending on he a ge o hei in luence, i.e., DI-SPME 205
ex ac ion, On-Fibe De i a iza ion, Fibe Deso p ion and Ca y-O e a oidance (Table 206
S2). A e wa ds, echnical limi a ions o his inno a i e me hodology we e poin ed ou , 207
which na owed down o 6 he numbe o pa ame e s admi ing u he op imiza ion. 208
None heless, 4 o hem, i.e., ibe coa ing, sample Ionic s eng h, sample pH and 209
de i a iza ion empe a u e could easily be op imized by a one- ac o -a -a- ime 210
app oach as hey a e disc e e a iables o o he wise consolida ed e e ences exis in 211
he scien i ic li e a u e which d as ically delimi s he ange o a ia ion. E en ually, 212
only wo pa ame e s emained as signi ican , ex ac ion and de i a iza ion imes, and 213
in need o u he op imiza ion. Thus, a esponse su ace me hodology (RSM), 214
consis ing o a ull ac o ial 22 wi h a cen al poin epea ed i e imes and ex ended 215
wi h 4 s a poin s, was applied o hem. Thus, a se o 13 expe imen s was andomly 216
pe o med. A e wa ds, he so wa e S a g aphics Cen u ion XVII was used o p ocess 217
he acqui ed expe imen al da a and ma hema ically i i o a second o de polynomial 218
model h ough he leas squa es me hod. 219
220
3 Resul s and discussion 221
8
3.1 Analy ical me hod de elopmen and op imiza ion o sewage and sludge 222
A selec ion o 22 PPCPs, in pa icula , 5 pha maceu icals and 2 o hei TPs as 223
well as 14 pe sonal ca e p oduc s and 1 o hei TPs, we e ini ially chosen as a ge 224
analy es. 225
The p o ocol ollowed o de elop and op imize he analy ical me hod, including 226
an expe imen al design, is desc ibed in he supplemen a y da a SD ile. In b ie , a e 227
he GC-MS leg was de eloped, he sample p e ea men pa o he me hodology was 228
op imized. Hence all he pa ame e s wi h a ole du ing he Online DI-SPME – On-Fibe 229
De i a iza ion we e iden i ied and some echnical limi s we e se . A e wa ds, some 230
p elimina y expe imen s we e ca ied ou in a one- ac o -a -a- ime app oach o 231
op imize he Type o Fibe Coa ing, Sample Ionic S eng h, Sample pH and 232
De i a iza ion empe a u e. Finally, as he ex ac ion and de i a iza ion ime could 233
in e e e wi h each o he , a esponse su ace me hod was designed based on a ull 234
ac o ial 22 wi h a cen al poin epea ed i e imes and ex ended wi h 4 s a poin s. 235
TS/N was selec ed as he esponse a iable du ing he op imiza ion, in o de o ge a 236
comp omise among he pe o mance o all he compounds. As a esul , he op imum 237
alue o he esponse a iable ob ained co esponded o an ex ac ion ime o 120 238
min and a de i a iza ion ime o 48 min. Tha is g aphically shown in Figu e 1. 239
A e he op imiza ion, en o he ini ial a ge PPCPs, including he analgesics 240
ace aminophen and ace ylsalicylic acid, he lipid egula o a o as a in, and he 241
an ibio ics amoxicillin, cip o loxacin, le o loxacin, no loxacin, sul ame hoxazole, 242
e y h omycin and cla i h omycin u ned ou o be unsui able o hei analysis by 243
Online DI-SPME – On-Fibe De i a iza ion – GC-MS, as hey exhibi ed a e y weak o 244
e en no esponse wha soe e . The e o e, hey we e uled ou and no included in he 245
me hod. 246
The inal me hods, which allowed o he analysis o 12 PPCPs including 3 TPs, 247
a e summa ized in Sec ions 2.2 and 2.3. Rep esen a i e SIM ch oma og ams, ob ained 248
om MilliQ® wa e and sewage sludge samples spiked wi h he a ge PPCPs a 4 µg L-1 249
and 400 ng g-1, espec i ely, using he op imized me hod condi ions, a e illus a ed in 250
Figu e 2. 251
252
253
9
3.2 Me hod alida ion and applica ion 254
3.2.1 Me hod alida ion 255
Se e al egula o y bodies (like he Uni ed S a es Food and D ug Adminis a ion 256
(FDA) [17] o Eu achem [18]), s anda diza ion agencies (like he In e na ional 257
Associa ion o O icial Analy ical Chemis s (AOAC In e na ional) [19]), and wo king 258
g oups and commi ees (like he Food and Ag icul u al O ganiza ion/Wo ld Heal h 259
(FAO/WHO) [20]) ha e published guidelines and equi emen s o me hod alida ion. 260
In addi ion, he Eu opean Union adop ed a decision [21] implemen ing a di ec i e 261
conce ning he pe o mance o analy ical me hods and he in e p e a ion o esul s. I 262
e e s o animal p oduc s. Howe e , i has been widely used as an illus a i e e e ence 263
in he design o cus omized alida ion p o ocols o en i onmen al analysis like in [22-264
25], as well as in he p esen s udy because o he lack o speci ic guidelines. 265
Hence, i e alida ion pa ame e s, i.e., accu acy, ME, p ecision, sensi i i y and 266
dynamic ange we e de e mined o all 12 a ge analy es included in he me hod 267
(clo ib a e, 1,4-benzoquinone, me hylpa aben, e hylpa aben, clo ib ic acid, ibup o en, 268
p opylpa aben, salicylic acid, p-hyd oxybenzoic acid, nap oxen, iclosan, diclo enac) in 269
sewage and sludge. In addi ion, a ca yo e es was also pe o med o ensu e he 270
absence o con amina ion be ween samples du ing he ins umen al leg o he 271
analysis. Two meaning ul le els o concen a ion pe ma ix −100 and 1000 ng L-1, and 272
100 and 400 ng g-1− ypical o he a ge compounds in eal sewage and sludge 273
samples, espec i ely, we e es ed o he ou i s pa ame e s, as ecommended by 274
[23, 24]. Each es was un in iplica e wi h he op imized me hod. The esul s, 275
a e age o bo h concen a ion le els, which a e discussed below, a e shown in Table 276
S4. 277
278
1) Accu acy: Absolu e eco e ies (%) we e de e mined by compa ing he 279
peak a eas ob ained om spiked samples analyzed using he op imized me hods wi h 280
he a eas ob ained om di ec injec ions (2 µL) o equi alen amoun s o s anda ds in 281
EA solu ions. As bo h sewage and sludge can con ain some o he a ge compounds, 282
non-spiked samples we e also analyzed and he peak a eas we e a e wa ds 283
sub ac ed om he spiked samples in o de o calcula e he absolu e eco e y. Table 284
S4A shows e y a iable absolu e eco e ies o sewage. Hence, SPME suppo ed good 285
16
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Analy ical Chemis y 83(12) (2011) 4579-4613. 475
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Comp ehensi e Sampling and Sample P epa a ion2012, pp. 419-460. 481
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con igu a ions o solid-phase mic oex ac ion and hei applica ions in ood and 483
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553
554
555
556
557
558
559
560
561
562
563
564
565
566
567
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Figu e 1: Response su ace a e applying an expe imen al design 22 + s a + 5 cen al 568 poin s 569
570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597
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Figu e 2: Ch oma og ams ob ained om A) 4000 ng L-1 MilliQ wa e and B) 400 ng g-1 598 sludge samples a e he op imized me hods we e applied 599 A) 600
601 602 B) 603
604
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Table 1: MS pa ame e s o he inal a ge compounds and in e nal s anda ds
1IS
Analy e
Chemical Name
Acquisi ion
window # 2 R (min) 3SIM ions, m/z
1
Clo ib a e
1
13.17
128
130
169
2
1,4-Benzoquinone
13.70
281
338
282
3
Me hylpa aben
2
15.23
209
210
135
4
Ace ylsalicylic acid
15.28
195
237
135
5
E hylpa aben
16.02
223
224
151
6
Clo ib ic acid
16.02
143
271
185
7
Ibup o en
16.38
263
264
117
1
Ibup o en-d3
16.34
266
267
164
8
P opylpa aben
17.04
237
238
151
2
P opylpa aben-d7
16.96
244
245
152
9
Salicylic acid
17.50
309
310
195
3
Salicylic acid-d4
17.45
313
314
312
10
Ace aminophen
3
18.24
208
265
166
11
P-hyd oxybenzoic acid
18.91
309
265
310
12
Nap oxen
4
21.07
287
185
288
4
Nap oxen-d3
20.95
290
188
207
13
T iclosan
21.66
347
345
200
5
T iclosan-d3
21.54
350
348
200
14
Diclo enac
5
23.73
352
214
354
6
Diclo enac-d4
23.73
356
218
358
1IS: In e nal S anda d; 2 R: Re en ion Time; 3SIM: Selec ed Ion Moni o ing

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