53 2 Advances in the Analysis of Pharmaceuticals in the Aquatic Environment Sandra Pérez and Damià Barceló 2.1 INTRODUCTION Recently, the focus of environmental analysis has shifted from the classic contami- nants, such as the persistent organic pollutants, toward the “emerging contaminants” detected recently in many environmental compartments. 1 Emerging contaminants aredenedascompoundsthatarenotcurrentlycoveredbyexistingregulations ofwaterquality,havenotbeenpreviouslystudied,andarethoughttobepotential threats to environmental ecosystems and human health and safety. In particular, the compounds that are being addressed include pharmaceuticals, drugs of abuse, and personal-care products. 2 The high water solubility of these organic compounds makesthemmobileintheaquaticmedia,hencetheycanpotentiallyinltratethesoil and then reach groundwater. Eventually, these compounds may nd their way into the drinking water supplies. In recent years the increasing use of drugs in farming, aquaculture, and human health has become a growing public concern because of their potential to cause undesirableecologicalandhumanhealtheffects.Themainconcernregarding Contents 2.1 Int roduction 53 2.2 Multiresidue Methods 56 2.3 Determination of Drugs According to Their Class 67 2.3.1 Analgesics and Antiinammatory Drugs 67 2.3.2 Antimicrobials 68 2.3.3 Ant iepilept ics, Blood Lipid Regulator s, a nd Psych iatr ic D rugs 70 2.3.4 Antitu moral Dr ugs 72 2.3.5 Cardiovascular Drugs (C-Blockers) and C 2- Sympathomimetics 72 2.3.6 Estrogens 72 2.3.7 X-Ray Contrast Agents 73 2.3.8 Drugs of Abuse 74 2.3.9 Other Drugs 75 2.4 Conclusion 75 Ack nowledgments 76 References 76 © 2008 by Taylor & Francis Group, LLC 54 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems pharmaceuticals is that they are being introduced continuously into water bodies aspollutants,andduetotheirbiologicalactivitythiscanleadtoadverseeffectsin aquatic ecosystems and potentially impact drinking water supplies. 3 Antibiotics are oneofthemostproblematicgroupsofpharmaceuticals,sincetheirincreasingusefor more than four decades has led to the selection of resistant bacteria that can threaten the effectiveness of antibiotics for the treatment of human infections. 4 Another group that has caused environmental concern is contraceptives and other endocrine disrup- to rs; due to their endocrine properties they can induce the feminization or masculin- iz ation in aquatic organisms. 5 In general the short- and long-term ecotoxicological effectsofpharmaceuticalsonwildlifehavenotyetbeenstudiedsufciently. Prescription and over-the-counter drugs have probably been in the environment foraslongastheyhavebeenused,butonlyrecentlyhaveanalyticalmethodsbeen developedtodetectpharmaceuticalsattracelevels. 6 Duetothedilutionandpossible degradation of these substances in the environment, low levels can be expected. Therefore, an analyte preconcentration procedure is almost always necessary in order to achieve desired levels of analytical sensitivity, often requiring high enrich- me ntfactors,between100and10,000.Suchenrichmentfactorsfordruganalysis are usually achieved using solid-phase extraction (SPE). Sensitive detection meth- ods such as gas chromatography-mass spectrometry (GC-MS), GC-tandem mass spectrometry (GC-MS/MS) or liquid chromatography-mass spectrometry (LC-MS), and LC-tandem mass spectrometry (LC-MS/MS) are also crucial for the analyti- cal determination of drugs in the environment. The main drawback of GC for drug analysis,however,isthatthistechniqueislimitedtocompoundswithhighvapor pressure. Since most drugs are polar substances, they need to be derivatized prior to injection in the GC. For this reason, the combination of atmospheric pressure ion- iz ation-MS (API-MS) with separation techniques such as LC or ultra performance liquid chromatography (UPLC) has become the method of choice in drug analysis. LCwithasinglequadrupoleMSanalyzeroffersgoodsensitivity,butwhenvery complex matrices such as raw sewage are investigated, insufcient selectivity often impairs the unequivocal identication of the analytes. Tandem MS affords superior performance in terms of sensitivity and selectivity in comparison with single quad- ru pole instruments. Liquid chromatographic techniques coupled to tandem MS or hybrid mass spectrometers with distinct analyzers such as triple quadrupole (QqQ), time-of-ight (ToF), quadrupole time-of-ight (QqToF), quadrupole ion trap (IT), and recently the quadrupole linear ion trap (QqLIT) are the most widely used instru- me ntal techniques for drug analysis. 7 Most of the data on the presence on pharmaceuticals in wastewaters, rivers, and drinking water come primarily from European studies 8,9 followed by those carried outintheUnitedStates. 10 These substances that are used in human and veterinary medicinecanentertheenvironmentviaanumberofpathwaysbutmainlyfrom discharges of wastewater treatment plants (WWTPs) or land application of sew- age sludge and animal manure, as depicted in Figure 2.1. Most active ingredients of pharmaceuticals are transformed only partially in the body and thus are excreted as a mixture of metabolites and bioactive forms into sewage systems. Therefore, the treatment of wastewaters in WWTPs plays a crucial role in the elimination of phar- ma ceuticalcompoundsbeforetheirdischargeintorivers.Duringtheapplicationof © 2008 by Taylor & Francis Group, LLC Advances in the Analysis of Pharmaceuticals in the Aquatic Environment 55 primary and secondary treatments, pharmaceutical compounds can be eliminated by sorption onto the sludge or microbially degraded to form metabolites that are usuallymorepolarthantheparentdrugs. 11 Inmanycasesthehighpolaritycombined with the low biodegradability exhibited by some pharmaceutical compounds results in inefcient elimination in WWTPs. The removal efciencies vary from plant to plantanddependonthedesignandoperationofthetreatmentsystems. 12,13 Thus, the majorsourceofpharmaceuticalresiduesdetectableinsurfacewatersaredischarges from WWTPs. Several studies reported the occurrence of pharmaceuticals at levels uptotheμg/Lrangeinrivers,streams,lakes,andgroundwater. 1,14 Rese archershaveyettodeterminetheoccurrence,fate,andpossibleeffectsof themostfrequentlyconsumeddrugsandtheirmainmetabolitesintheaquaticenvi- ro nment. Exceptionally high levels of drugs have been reported—for example, the occurrenceoftheantiasthmadrugsalbutamolinwaterfromthePoRiver. 15 The researchersconcludedthattheirdatareectedtheillegaluseofsalbutamolbylocal farmers to promote growth in cattle. The determination of pharmaceuticals and drugs of abuse in the environment by appl yingthep rinciple that what goes in must come outcanbeahelpfultooltoestimatethedrugconsumptionintheinvestigatedareas. For example, Italian researchers measured the levels of benzoylecgonine, the major urinarymetaboliteofcocaine,inwastewaterfromseveralItaliancities. 16 What they foundwassurprising:cocaineuseappearedtobefarhigherthanthepublichealth ofcials previously thought. This review provides an overview on analytical protocols used in determining drugs and some of their metabolites in aqueous and solid environmental samples.  #! #! !"!  $" #!!  #" #!" ! &  #! %# # "   "   #%" #  & "  & "  & "   !"   %" " !   #"$!"  #! (  ( #" "" ! !  #! "  '!   !"%"  """!  FIGURE 2.1 Pathways of pharmaceuticals and their metabolites in the environment. © 2008 by Taylor & Francis Group, LLC 56 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems Technologicalprogressintheeldsofsampleextractionanddetectionbymassspec- trometry (MS) techniques (hybrid and tandem mass spectrometers) for analyzing antibiotics, antiinammatory/analgesics, lipid regulating agents, psychiatric drugs, sedatives, iodated X-ray contrast media, diuretics, drugs of abuse, and some human metabolitesintheaquaticenvironmentarediscussed. 17–19 The recent trends in mul- tiresidue methodologies for the determination of the drugs and their human metabo- li teswillbereviewedhere. 2.2 MULTIRESIDUE METHODS Manyanalyticalmethodologiesforthedeterminationofdrugsinwaterbodiesfocused on selected therapeutic classes. Multiresidue methods, however, are becoming more widespreadinresponsetotheneedofmonitoringawiderangeofpharmaceuticals that belong to diverse drug classes in wastewater, surface water, and groundwater. Thelatterapproachoffersadvantagesintermsofprovidingamorecomprehensive picture of the occurrence and fate of the contaminants in the environment. In addi - ti on,thesimultaneousdeterminationofalargenumberofanalytesbyasinglemethod represents a less time-consuming and hence more economical approach as compared with applying class-specic analytical protocols. The multiresidue methods found in the literature are diverse, with target analytes being selected commonly on the basis of their consumption in the country where the study is being conducted, the rate of metabolism of drugs, the environmental occurrence, and persistence in the environment. In this chapter, multiresidue methods for drug analysis in the aquatic environment are reviewed. Ternes et al. 20 reported the determination of neutral pharmaceuticals: propyphen- azone (analgesic), phenylbutazone (antiinammatory), diazepam (psychiatric drug), omeprazole (antiulcer), nifedipine (calcium antagonist), glibenclamide (antidiabetic), and two human metabolites: 4-aminoantipyridine (metabolite of metamizole) and oxyphenbutazone (metabolite of phenylbutazone) with a multiresidue methodology including a one-step extraction method based on SPE with RP-C 18 material eluted with methanol. The analysis was performed by LC with detection by electrospray ionization(ESI)tandemMSinmultiplereactionmonitoring(MRM)mode,which is the acquisition mode providing the best sensitivity and selectivity for quantitative analysis. Low limits of detection and reasonable recoveries for the selected drugs in differentmatriceswereachieved.Thiswork 20 investigated the losses in the recover- iesofsomedrugsduetomatriximpurities,whicheitherreducedthesorptionef- ci encies on the C 18 material or led to signal suppression in the ESI interface. The authors spiked inuent wastewater extracts with the target analytes and found that the recoveries were not appreciably higher in comparison to the recoveries over the total method. Consequently, the signal suppression in ESI played a decisive role in the losses of 4-aminoantipyrine, omeprazole, oxyphenbutazone, phenylbutazone, and propyphenazone. For most of the compounds, compensation for the losses was achieved by addition of the surrogate standard 10,11-dihydrocarbamazepine. How - ev er, low corrected recoveries for oxyphenbutazone, phenylbutazone and 4-amino- © 2008 by Taylor & Francis Group, LLC Advances in the Analysis of Pharmaceuticals in the Aquatic Environment 57 antipyridine indicated that the determination of these compounds was still rather semiquantitative. 20 Vanderford et al. 21 developed an analytical method for the determination of 21 pharmaceuticals in water, choosing them based on their occurrence in the environ- m e nt and their dissimilar structural and physicochemical structures. They used also a one-step extraction method employing SPE with a hydrophilic-lipophilic balance (HLB). The separation and detection was performed with LC-MS/MS, using ESI in either positive or negative mode or atmospheric pressure chemical ionization (APCI) inpositivemode.Theanalyticalmethodprovidedasimpleandsensitivemethodfor the detection of a wide range of pharmaceuticals with recoveries in deionized water above 80% for all of the compounds. The authors also studied the effect of sample preservatives on the recovery of the pharmaceuticals. 21 They compared formalde- hyde and sulfuric acid, obtaining the best results for the latter, which prevented the degradationofthetargetcompoundsanddidnotadverselyaffecttheirrecoveries. Matrixeffectswerealsoexaminedinthisworkshowingthatallcompoundsdetected with(+)ESIand(–)ESI,excepthydrocodone,showedaconsiderabledegreeofion - ization suppression. Hydrocodone, though, showed signal enhancement. In another workthesamegroupreportedamethodologyusinganisotopedilutiontechniquefor every analyte to compensate for matrix effects in the ESI source, SPE losses, and instrument variability. 22 The method was tested with three matrices (wastewater, surface water, and drinking water), and the results indicated that the method was very robust using isotope dilution for each target compound. The work described a method that analyzed 15 pharmaceuticals and 4 metabolites using SPE (HLB) cou - p l ed with LC-MS/MS with ESI source. Matrix spike recoveries for all compounds were between 88 and 106% for wastewater inuent, 85 and 108% for wastewater efuent, 72 and 105% for surface water impacted by wastewater, 96 and 113% for surfacewater,and91and116%fordrinkingwater.Themethoddetectionlimits werebetween0.25and1ng/L. Astudy 23 evaluated different strategies to reduce matrix effects in LC-MS/MS withanESIinterface.First,thepeakareaofthetargetcompoundsinsolventwere compared with the target compounds spiked in matrix extracts obtaining signal sup - pressions in the range of 40 to 90%. Next, internal calibration curves with inter- n a l labeled standard in solvent and in spiked matrix extracts were prepared. The overlapping of both curves conrmed that signal losses experienced by the analytes were corrected by the internal standards. Finally, the effectiveness of diluting sample extracts was studied. For this purpose, the signals obtained after sequential dilution of a WWTP efuent and inuent extract were compared with the ones obtained for the corresponding concentrations of the standards in the solvent. The authors consid - eredthatmatrixeffectswereeliminatedwithdilutions1:2and1:4,andthisapproach wasselectedforthiswork.Fortheextractionofthetargetanalytes,one-stepSPE testing Oasis HLB, Isolute ENV+ and Isolute C 18 with and without sample acidi- cationwasoptimized.OasisHLB,withsorbentbasedonahydrophilic-lipophilic polymer, provided high recoveries for all target compounds at neutral pH. Recoveries were higher than 60% for both surface and wastewaters, with the exception of sev - e r al compounds: ranitidine (50%), sotalol (50%), famotidine (50%), and mevastatin (34%). © 2008 by Taylor & Francis Group, LLC 58 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems Miao et al. 24 reportedamethodusingSPE(C 18 )andLC-(-)-ESI-MS/MSforthe simultaneousdetectionofnineacidicpharmaceuticaldrugs(bezabrate,clobric acid, diclofenac, fenoprofen, gembrozil, ibuprofen, indomethacin, ketoprofen, and naproxen) in WWTP efuents. The recoveries ranged from 59% (indomethacin) to 92%(fenoprofen)intheWWTPefuent.Thespecicityofthemethodwaschecked spiking samples with analytes at concentration of 0.05 μg/L. Two interfering peaks resulting from endogenous components in the WWTP efuent were detected in the MRM channels for fenoprofen and indomethacin. Coextractives in the WWTP yielded fragmentation patterns similar to fenoprofen and indomethacin. However, the separationefciencyprovidedbyhighperformanceliquidchromatography(HPLC) was sufcient to resolve these interfering compounds from the analytes showing the importanceofusingLCtoimprovetheselectivityoftheanalysis. AmultiresidueanalyticalmethodusingSPEandLC-MS/MSfor28pharma - ceuticals including antimicrobial drugs, two diuretics (furosemide and hydrochlo- r o thiazide), cardiovascular (atenolol and enalapril), antiulcer, psychiatric drugs, an antiinammatory, a C 2 -sympathomimetic (salbutamol), a lipid regulator, some estro- gens (17C- estradiol, 1 7B-ethinylestradiol, estrone), two antitumorals (cyclophospha- mide and methotrexate), and two metabolites (clobric acid and demethyl diazepam) was developed. 25 To optimize the extraction method, several stationary phases and different PH samples were tested. The cartridges were Oasis MCX at pH 1.5/2.0 and3.0forallthecompoundsandatpH7.0/7.5foromeprazole;LiChroluteENat pH 3.0, 5.0, 7.0, and 9.0 for all the compounds; Bakerbond C 18 at pH 8.0 and 9.5 for the extraction of amoxicillin;a nd O asisHLBatpH7.0foromeprazoleandpH 8.5/9.0 for amoxycillin. They selected Oasis MCX for water samples at pH 1.5/2.0 and LiChrolute EN for water samples at pH 7. Recoveries of the pharmaceuticals weremostlygreaterthan70%andinstrumentalandmethodlimitsofdetectionin the order of ng/L. Vieno et al. 26 developedamethodthatallowedthequanticationofthefourC- blockers—acebutolol, atenolol, metoprolol and sotalol, carbamazepine—and the three uoroquinolones antibiotics—ciprooxacin, ooxacin, and noroxacin—in ground - water,surfacewaters,andrawandtreatedsewages.Theauthors 26 studied the effect ofthewashingandoftheelutingsolventandpHontheextractionstepusingasingle pretreatment (SPE, Oasis HLB). Prior to the elution step, the adsorbent was washed with2mLof5%ofmethanolin2%aqueousNH 4 OH showing improvements in the MS detection in terms of decreased ion suppression and thus improved detectability of the compounds. Methanol was the solvent of choice yielding the highest recoveries ascomparedwiththoseobtainedwithacetonitrileoracetone.Thestudyoftheinu - e n ceofthepHofthewaterintheextractionmethodologywasperformedatthree pH values: 4.0, 7.5, and 10.0. For most of the compounds, the pH did not have a pro - nounced effect on the recovery, with the exception of atenolol and sotalol, which were poorlyrecoveredatlowpH(<10%atpH4.0).ThesampleswereanalyzedwithLC- MS/MSusingESIinpositivemodeshowingionsuppressionintheESIsource. 26 To evaluate the matrix effects, the authors infused continuously a standard solution into themassspectrometerandtheninjectedeithersolventorarealsampleextractonto theLCcolumn.Moreover,SPEextractsofgroundwater,surfacewater,andwastewa - t e rinuentandefuentwerespikedwithpharmaceuticals,andspikedsampleswere © 2008 by Taylor & Francis Group, LLC Advances in the Analysis of Pharmaceuticals in the Aquatic Environment 59 analyzedinLC-MS/MSwithESIinterface.Noionsuppressionwasnoticedforany of the analytes in groundwater extracts; some signal suppression (<8%) was noticed for sotalol, acebutolol, and metoprolol for the surface-water extract; and more severe signal suppression (40%) was observed in the wastewater inuents and efuents. The authors 26 reported relative recoveries higher than those reported previously by other authors 24 showinganimprovementofthegeneralmethodologyforallthetargetana- lytes, except for ciprooxacin and noroxacin. The determination of selected drugs and their metabolites with a multiresidue methodology was also reported by Zuehlke et al. 27 Carbamazepine, dimethylami- nophenazone, phenazone, propyphenazone, 1-acetyl-1-methyl-2-dimethyloxanoyl-2- phenylhydrazide, 1-acetyl-1-methyl-2-phenylhydrazide, two human metabolites of metamizole (formylaminoantipyridine and aminoantipyridine), and two micobio - l o gical metabolites (1,5-dimethyl-1,2-dehydro-3-pyrazolone, 4-(2-methylethyl)-1,5- dimethyl-1,2-dehydro-3-pyrazole) were studied. To allow for efcient SPE of the two microbiological metabolites from water on a conventional C 18 sorbent, the authors 27 prepared the water samples by a simple in situ d erivatization w ith acetic anhy- drideinbasicmediainordertodecreasethepolarityandtoincreasethemolecular weight of these substances by acetylation. Only the two analytes 1,5-dimethyl-1,2- dehydro-3-pyrazolone and 4-(2-methylethyl)-1,5-dimethyl-1,2-dehydro-3-pyrazole werederivatizedwhiletheothercompoundswerequantitativelyextractedwithout chemical transformation. The analytes were then separated by LC-APCI-MS/MS and quantied by comparison with the internal standard, dihydrocarbamazepine. 27 AlthoughESIledtohigherpeakintensitiesthanAPCI,thelatterinterfacewascho- s e n because it provided a matrix-independent ionization resulting in recoveries of ~100% (Table 2.1). Al thoughtheuseofGC-MSgenerallyrequiresthederivatizationofpolardrugs, Boydetal. 28 usedthisapproachtoanalyzeacetaminophen,uoxetine,ibuprofen, naproxen,andclobricacid,ahumanmetaboliteofclobrateandetobrate.Thetar- g e tcompoundswereisolatedfromwastewater,surfacewater,anduntreateddrink- i n gwatersamplesbySPEusingapolarSDB-XCEmporedisk.Derivatizationwith N,O -bis(trimethylsilyl)-triuoroacetamideinthepresenceoftrimethylchlorosilane wasusedtoenhancethermalstabilityofclobricacid,whichthermallydegradedin theGCinjectionport,andtoreducethepolarityofspecictargetanalytes(clobric acid,ibuprofen,andnaproxen)inordertofacilitatetheirGC-MSanalysis.Thelimits ofdetectionwerebetween0.6(clobricacid)and25.8ng/L(uoxetine).Although therecoveriesformostofthecompoundsweregreaterthan47%(Table2 .1), a cet- am inophen was repeatedly not detected possibly due to the weak retention of this compound on the extraction disk. Next, analytical methods using multiple extraction methods, different liq - u i d chromatography eluents, or the combination of two detection techniques are reviewed. 10,29,30 Sacher et al. 29 reported the determination of 60 pharmaceuticals including analgesics, antirheumatics, C-blockers, b roncholitics, lipid regulators including two metabolites, antiepileptics, vasodilators, tranquilizers, antitumoral drugs, iodated X-ray contrast media and antimicrobials in groundwater with dif - ferentSPEprocedures,andthecombinationofGC-MS(afterderivatizationofthe acidic compounds) and LC-ESI-MS/MS. Different stationary phases, pH (3, 5, and © 2008 by Taylor & Francis Group, LLC 60 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems TABLE2.1 Methods for the Analysis of Drugs in Aqueous Environmental Samples Analytes Matrix Extraction Procedure (for SPE: Sorbent, Sample pH; Elution Solvent(s)) Separation and Detection Method Recovery [%] Limit of Detection and Quantification Ref. Multiresidue method for neutral drugs and 2 metabolites GW 1 ,SW 2 , WW 3 SPE 4 : Isolute C 18 , pH 7–7.5; MeOH LC-(+)ESI-MS/MS GW>80 SW and WW: 9–97% MQL 5 GW:10 ng/L WW: 25–250 ng/L [20] Multiresidue method for neutral and acidic drugs SW, WW SPE: HLB, pH 2; MeOH LC-(+/–)ESI-MS/MS Deionized water: 80 IDL 6 :12–32 ng/L [21] Multiresidue method for 15 drugs and 4 human metabolites DW7, SW, WW SPE: HLB; MeOH/MTBE (1:9) LC-(+/–)ESI-MS/MS 90–110 MDL: 0.25–1 ng/L [22] Multiresidue method for neutral and acidic drugs and 5 metabolites SW, WW SPE: HLB, pH 7; MeOH [Optimization of stationary phases and pH] LC-(+/–)ESI-MS/MS SW: 50–116 WW: 60–102 MDL 8 SW:1–30 ng/L WW:3–160 ng/L [23] Multiresidue method for neutral and basic drugs GW, SW, WW SPE: HLB, pH 10; MeOH [Optimization of washing and eluting solvent, and of pH] LC-(+)ESI-MS/MS GW: 50–119 SW: 22–113 WW: 64–115 IQL 9 : 0.46–10.6 μg/L MQL GW:1–10 ng/L, SW: 1–24 ng/L, WW: 1.4–29 ng/L [26] Multiresidue method for neutral and acidic drugs and 5 metabolites GW, SW, WW SPE: C 18 ; MeOH LC-(+)APCI-MS/MS Interface optimization (ESI and APCI) GW, SW and WW:87– 117 except for dimethylaminophenazone MQL: 10–20 ng/L [27] Multiresidue method for acidic drugs WW SPE: LiChrolute 100 RP-18, pH 2; MeOH LC-(–)ESI-MS/MS 59–92 MDL: 5–20 ng/L [24] © 2008 by Taylor & Francis Group, LLC Advances in the Analysis of Pharmaceuticals in the Aquatic Environment 61 Multiresidue method for 28 drugs and 2 metabolites WW SPE: 1 Oasis MCX, pH 1.5/2.0; MeOH, MeOH (+2% NH 3 ) and MeOH (+0.2% NaOH) 2. LiChrolute EN, pH 7; MeOH, EtOAc [Optimization of stationary phases and pH] LC-(+/–)ESI-MS/MS >70 IQL: 600–39400 ng/L MQL:0.1–5.2 ng/L [25] Multiresidue method for 4 drugs and 1 human metabolite DW, SW, WW SPE: SDB-XC, pH 2; MeOH, CH 2 Cl 2 /MeOH GC-MS 47–88 IDL:0.6–25.8 ng/L [28] Combined method for 60 drugs and their metabolites SW SPE: RP C 18 pH 3, PPL Bond-Elut pH 7, LiChrolute EN pH 3, Isolut ENV+ pH 5; MeOH, acetonitrile, water, triethylamine GC-MS and LC-(+)ESI-MS/MS 36–151 IDL: 1.8–13 ng/L [29] Combined method for 11 drugs and 2 metabolites SW, WW SPE: Strata X, pH 3; MeOH [Stationary phase optimization] LC-(+/–)ESI-IT-MS >60 Except for lofepramine and mefenaminic acid MQL: 10–50 ng/L [30] Combined method for 11 drugs and 2 metabolites SW, WW SPE: Strata X, pH 3; MeOH, MeOH (+2% HOAc), MeOH (+2% NH 3 ) [Stationary phase optimization] LC-(+/–)ESI-IT-MS >60 Except for chloroquine and chlosantel MDL: 1–20 ng/L IQL: 20–105 pg [32] Combined method for 13 drugs SW, GW, DW SPE: Oasis-MCX, pH 3; MeOH/NH 3 (19:1) LC-(+/–)ESI-MS/ MS and LC-QqToF-MS 60–75 except for fenobrate (36) MQL: 5–25 ng/L [33] (Continued) © 2008 by Taylor & Francis Group, LLC 62 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems TABLE 2.1 (Continued) Analytes Matrix Extraction Procedure (for SPE: Sorbent, Sample pH; Elution Solvent(s)) Separation and Detection Method Recovery [%] Limit of Detection and Quantification Ref. Combined method for >50 drugs and their metabolites SW SPE: 1:tandem HLB and MCX; MeOH (+5% NH 3 ) 2:HLB; MeOH LC-(+)ESI-MS >80 -10 [10] Combined method for >10 drugs and 3 human metabolites SW, Seawater SPE: Oasis HLB; hexane, EtOAc and MeOH LC-(+)ESI-MS/MS and GC-MS 70–100 MQL:0.07–0.69 ng/L [34] Combined method for 5 neutral ad acidic drugs and 1 metabolite SW, WW SPE: Oasis HLB; EtOAc/ acetone (1:1)[Optimization of washing and eluting solvent] GC-MS 71–118 MDL 1–10 ng/L [35] Methodology by therapeutic class Acidic drugs, acetylsalicylic acid and 4 metabolites SW, DW, WW SPE: C18, pH 2; MeOH GC-MS and GC-IT-MS/MS 58–90 MQL: WW: 50–250 (GC-MS) SW:5–20 (GC-MS) DW: 1–10 (GC-IT-MS/MS) [36] 8 acidic drugs SW SPE: C18, pH 2; MeOH GC-IT-MS/MS ≤90 MQL:1 ng/L [38] 5 acidic drugs GW, SW, WW SPE: Oasis MCX, pH 2; acetone LC-(–)ESI-MS/MS GW: 82–103 SW: 75–112 WW: 57–100 MQL:1–25 ng/L [39] © 2008 by Taylor & Francis Group, LLC [...]... determination by GC-MS and GC-IT-MS/MS The MQL down to 10 ng/L were achieved in wastewater effluents as well in river water by GC-MS and down to 1 ng/L using GC-IT-MS/MS Other authors also used GC-IT-MS/MS for the determination of eight acidic pharmaceuticals in water by SPE on RP-C18 In- port methylation in the GC using trimethylsulfoniumhydroxide improved the detection limits such that concentrations in the ng/L... to their basic character The protonated molecule is the selected precursor ion, and the most intense diagnostic ion is m/z 116 corresponding to [(N-isopropyl-N -2 - hydroxypropylamine)] Ternes et al48 reported the determination of several -blockers and 2- sympathomimetics in waters comparing two methods of separation and detection: GC-MS and LC-MS/MS For GC-MS, the sample preparation included SPE (C18-endcapped),... in drinking water and surface water and 50 ng/L in wastewater.48 The authors recommended the use of LC-(+)ESI-MS/MS for the analysis of these polar molecules in the environment, because the derivatization of the hydroxyl groups required for GC-MS analysis was incomplete 2. 3.6 ESTROGENS Recently, a multitude of chemicals have shown to act as endocrine disrupters disturbing the hormonal systems of aquatic... LC-(+)ESI-MS/MS LC-(+)ESI/MS-MS SW: 90–116 WW: 57–113 (ioxithalamic acid: 35) 55–100 MQL:10–50 ng/L [ 62] MDL: 50 ng/L [63, 64] [29 ] [69] Advances in the Analysis of Pharmaceuticals in the Aquatic Environment 6 ICM 65 © 20 08 by Taylor & Francis Group, LLC 66 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems 7), and eluting solvents were used for the determination of the target... total run of 11 min Matrix effects were also studied with four kinds of matrices, HPLC water, surface water, and influent and effluent from a WWTP Ion suppression was highest in the influent To correct ion suppression, 10,11-dihydrocarbamazepine was used as internal standard © 20 08 by Taylor & Francis Group, LLC 72 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems 2. 3.4 ANTITUMORAL... reported the determination of 13 antimicrobials, sulfonamides, and tetracyclines, in surface waters using SPE (Oasis HLB) and LC-(+)ESI-IT-MS but in CRM mode For the enrichment of the water samples, pH between 2 and 3 was used because tetracyclines are not stable at pH < 2 They checked the effect of the column diameter, flow rate, and temperature (15°C, 25 °C, 35°C) on the peak shape in the chromatographic... for ibuprofen, naproxen, ketoprofen, and diclofenac Nevertheless, in the WWTP effluent, the relative recovery of ibuprofen was only 57% and 67% for bezafibrate Low method limits of detection were reported for ibuprofen, diclofenac, and bezafibrate They were 1 ng/L in ground and surface waters and 5 ng/L in WWTP samples, and 5 and 25 ng/L for naproxen and ketoprofen, respectively (Table 2. 1) 2. 3 .2 ANTIMICROBIALS... suppression and losses in the SPE method, a method for 13 antimicrobials and the metabolite N4-acetyl-sulfamethoxazole in wastewater using five labeled internal standards was reported. 42 The method combined SPE (Oasis HLB) and LC-(+)ESI-MS/MS showing recoveries above 80% (Table 2. 1), with the exception of trimethoprim, where they ranged between 30 and 47%, probably because of the use of nonideal surrogate standard... 0.5–1 ng/ LWW: 1 2 ng/L IDL: C/MS: 1 20 ng/mL LC-ESI-MS: 0.1 20 ng/L LC-ESI-MS/MS: 0.1–10 ng/L MDL: 6.7 ng/L MQL: 20 ng/L Ref [54] [55] [48] [57] [58] [ 12] Fate of Pharmaceuticals in the Environment and in Water Treatment Systems TABLE 2. 1 (Continued) DW, SW, WW 4 ICM GW SPE: ENV+, pH 2. 8; MeOH [Optimization of stationary phases] SPE: ENV+ and Envi-Carb, pH 3.5; MeOH and acetonitrile /water (1:1) back... combination of LC-ESI-MS/MS and GC-MS after derivatization with methylchloromethanoate for the determination of selected pharmaceuticals, among them analgesics with emphasis on ibuprofen and its metabolites (hydroxy-ibuprofen and carboxy-ibuprofen), -blockers, antidepressants in wastewater and seawater, was reported The extraction procedure was performed in 6-mL glass cartridges with the same packing . of metamizole (formylaminoantipyridine and aminoantipyridine), and two micobio - l o gical metabolites (1,5-dimethyl-1 , 2- dehydro-3-pyrazolone, 4-( 2- methylethyl )-1 , 5- dimethyl-1 , 2- dehydro-3-pyrazole) were. groundwater. Thelatterapproachoffersadvantagesintermsofprovidingamorecomprehensive picture of the occurrence and fate of the contaminants in the environment. In addi - ti on,thesimultaneousdeterminationofalargenumberofanalytesbyasinglemethod represents. """!  FIGURE 2. 1 Pathways of pharmaceuticals and their metabolites in the environment. © 20 08 by Taylor & Francis Group, LLC 56 Fate of Pharmaceuticals in the Environment and in Water Treatment Systems Technologicalprogressintheeldsofsampleextractionanddetectionbymassspec- trometry