Bletsou, Anna

Digital Repository of Hellenic Managing Authority of the Operational Programme &quot;Education and Lifelong Learning&quot; (EDULLL)

Target screening of 2327 emerging pollutants in environmental samples by RP-LC-QTOF-HR-MS/MS with an accurate-mass database°Anna BletsouLaboratory of Analytical ChemistryDepartment of ChemistryUniversity of AthensEmerging Pollutants (EPs)• Pesticides• Pharmaceuticals• Illicit drugs• Personal care products• Endocrine disruptive compounds (EDCs)• Flame retardants• Food additives• Disinfection by-products +Metabolites     &Transformation Products (TPs)> 700 pesticides> 200 pharmaceuticals, illicit, DoA~ 300 steroids & doping compounds~ 100 compounds like industrial chemicals, food additives, dies and natural occurring compounds (aminoacids)~ 300 metabolites & TPsdatabase of 2327 EPsWastewater:Potentially tens of thousands of substancesWorkflowReportData ProcessingScreener bbCID DatabaseAccurate mass database:Qualifier Ions and RTIdentificationConfirmationSampleSample ExtractionLC-QTOFUHPLC + Bruker QTOFChromatographic separationwith accurate mass bbCID data acquisition 100 mL wastewater (GFF filtration) IS spiking (100 ng/L) SPE           Mixed-bed cartridges Extraction: Neutral, Basic & Acidic CompoundsStrata-XMixture:Strata-XCW,Strata-XAW,ENVI+ 100 times preconcentrationLocation: WWTP of Athens, GreecePeriod: March 2014Samples: 24-h composite flow-proportional samples ofinfluent wastewaters & effluent wastewatersover a weekSample Preparation:Sampling & Sample preparation*Kern et al. EST (2009) 43(18):7039Column: Acclaim RSLC 120 C18 2.2 μm, 2.1 × 100 mm Pre-column: VanGuard (Waters): Acquity UPLC BEH C18 1.7 μm, 2.1 × 5 mm  20 min chromatogram Gradient elution program in the M.P.  Gradient elution program in the flow rate Positive & Negative ESI bbCID mode(MS & MS/MS simultaneously)InstrumentationbbCID modeLow CE (4 ev) (pass all)MS spectraHigh CE (25 ev) (fragment all)MS/MS spectraMaXis ImpactUltra High ResolutionTime-of-Flight Mass SpectrometerRapidly alternating MS & MS/MS acquisitionsMSbbCID MS/MS  Confirmation Detection2224 compounds in (+) ESI580 compounds in (-) ESI Retention time Molecular Formula Adducts In-source fragments bbCID fragments Ion RatiosDatabaseOptimization-ValidationOptimization of the evaluation methodFindArea 1000 (+)/ 600 (-)Intensity 250(+)/ 150 (-)Scoringmin maxret. Time (min) 0.1 0.4accuracy (ppm) 2.5 5mSigma threshold 100 200validation dataset  195 compounds 10% of the compounds of the total database Representative retention time Representative physicochemical properties Representative ionization behavior Compounds from every class of EPsScreening method:The method used to identify the non-compliant samples (true positive) with a β-error less than 5%C spiked samples (μg/L)% ‘missed compounds due to Area / Intensity thresholdFalse Negative results(compounds)Successful Identification Rate (%)1 0 1 990.5 0 2 990.25 1.3 2 970.05 3.3 3 950.025 5.2 5 92Successful Identification Rate >95%Validation Calibration curves (solvent, matrix & spiked samples) (8 levels of concentration) Repeatability, recoveries and matrix effect The screening detection limit (SDL) and the limit of identification (LOI):• SDL: the lowest concentration level tested for which a compound was detected in all samples; 2 Identification Points (IPs)• LOI: the lowest concentration tested for which a compound was satisfactorily identified in all spiked samples; 4 Identification Points (IPs) Decision limit (CCα) & Detection capability (CCβ)Propose an HRMS oriented validation scheme,according to 2002/657/EC, SANCO 12571/2013 & literature Parameter Thresholds Identification Points (IP)Precursor ion (accuracy) +tR2 ppm 2.5Isotopic fit (ratio of M+1, M+2)200 mSigma 0.5Fragments (accuracy) 5 ppm 2.5Validation results% Recoveries % Matrix Effect91.7 % analytes signal suppression8.3 % analytes signal enhancement% Repeatability (n=6)94.5 % analytes <20% RSD05101520253035< 60% 60-80% 80-100% 100-120% > 120%% AnalytesScreening Detection Limits (SDL) –Limits of Identification (LOI)01020304050600.0025 0.005 0.01 0.025 0.05 0.25 0.5 1% AnalytesC (μg/L)SDLLOI01020304050<0.005 0.005-0.010.01-0.0250.025-0.050.05-0.250.25-0.50.5 - 1 >1% AnalytesC (μg/L)CCβDetection capability CCβResultsInfluent wastewaterEffluent wastewaterScreening:  ≥2IPs (tR + precursor ion)Identification: ≥4IPs (tR + precursor ion + fragment ion)Effluent wastewaters:301 compoundsInfluent wastewaters:338 compoundsScreened compounds219Identified compounds192188 109371 compounds detectedResults• 61 pesticides • 205 licit & illicit drugs• 4 sweeteners• 10 PFCs• 8 Aminoacids• 47 TPs• 51 pesticides• 191 licit & illicit drugs• 4 sweeteners• 11 PFCs• 4 Aminoacids• 49 TPs80.048789.0593102.0545114.1277116.0703134.0964+bbCID MS, 25.0eV, 4.80-4.82min #557-559, Background Subtracted, Background Subtracted0.00.51.01.55x10Intens.50 60 70 80 90 100 110 120 130 m/z135.0689136.0504137.0588138.0368140.1394141.0394+MS, 4.79-4.83min #556-560, Background Subtracted, Background Subtracted0500100015002000Intens.135 136 137 138 139 140 141 m/z136.1082137.1095138.0949 139.1260140.1468+MS, 4.78min #5540123454x10Intens.135.5 136.0 136.5 137.0 137.5 138.0 138.5 139.0 139.5 140.0 m/zScreening of 1-OH-BTRSample Spiked SampleMS spectraMS/MS spectraAccuracy 1.3 ppm: 2.5 IPmSigma: -2.5 IPsXIC (± 0.005mDa) 0.05 μg/L90.781891.041292.0228+bbCID MS, 25.0eV, 4.80-4.82min #557-559, Background Subtracted, Background Subtracted0500100015002000Intens.90.5 91.0 91.5 92.0 92.5 93.0 93.5 94.0 m/zno MS/MS spectra for the sampleResults112.1121129.1385242.0469370.1782+bbCID MS, 25.0eV, 5.25min #611, Background Subtracted, Background Subtracted0123455x10Intens.50 100 150 200 250 300 350 400 m/z370.1779+MS, 5.24-5.28min #610-614, Background Subtracted, Background Subtracted0123455x10Intens.350 375 400 425 450 475 500 525 m/z57.0335101.0598113.0294126.0369145.0860189.1115242.0478357.2118370.1798+bbCID MS, 25.0eV, 5.24-5.25min #609-611, Background Subtracted, Background Subtracted02465x10Intens.50 100 150 200 250 300 350 m/z370.1796374.2390379.1939388.2546+MS, 5.23-5.26min #608-612, Background Subtracted, Background Subtracted0.000.250.500.751.001.256x10Intens.360 370 380 390 400 410 m/zIdentification of AmisulprideSample Spiked SampleMS spectraMS/MS spectraAccuracy 0.1 ppm: 2.5 IP32 mSigma: 0.5 IPXIC (± 0.005mDa) 0.05 μg/L8 IPs2.5 IPs2.5 IPsResultsResultsQuantitation010203040< 1 ng/L 1-10 ng/L 10-100 ng/L100-1000 ng/L1000-10000 ng/L (1-10 μg/L)> 10000 ng/L (> 10 μg/L)% AnalytesConcentration rangeinfluenteffluent051015202530Benzoic acidIsoleucineMetforminGlutamic acid CaffeineTramadolClarithromycinVenlafaxineParacetamolDiethyl …Carbamazepine LamotrigineProlinamideMetoprololEthyl sulfateTolyltriazoleDi-n-butyl …Dimethachlo…AcamprosatAtenololallo-THF O-…CyclamateCitalopramRanitidineC (μg/L)Most abundant compoundsin effluent WW020406080100MetforminBenzoic acidValproic acidCyclamateAlanine Caffeineg-aminobutiric …TheophylineSalicylic acidParacetamolSaccharineClarithromycinRanitidineMethionine PrednisolonDiethyl phthalateAcesulfame DesvenlafaxineAtenololIopromidAtorvastatinSalbutamolIbuprofenCitalopramTramadolC (μg/L)Most abundant compounds in influent WWResults QuantitationResults PesticidesFluometuron, Azoxystrobin & Dimethachlor met.Influent: 0.3 ng/L- 2.04 μg/LEffluent: 0.2 ng/L- 13.6 μg/L DrugsMetformin →most abundant Haloperidol → less abundantInfluent: 0.14 ng/L - 93 μg/L Effluent: 0.5 ng/L - 35 μg/L SweetenersInfluent: 0.6 μg/L sucralose - 24 μg/L cyclamateEffluent: removal >60 % Other chemicalsBenzoic acid: 49 μg/L (influent) - 29 μg/L (effluent) Ethyl sulfate: 3.6 μg/L (influent) – 1.3 μg/L (effluent)  AminoacidsInfluent: Conc. >2 μg/L (8 AAs) Valine: most abundantEffluent: significant removal rates (4 AAs), Val & GABA > 10 μg/L QuantitationConclusions In-house database with information for 2327 compounds Mixed-bed SPE covers a wide range of compounds Optimization & validation of the target screening method HR-MS oriented validation scheme & IPs system Application in influent & effluent wastewater samples Screening, Identification & Quantification of the analytes Continuously increasing database with new reference standards → Retrospective target screening Dimitris DamalasKaterina PsomaPablo Gago Ferrero Nikolaos ThomaidisEmma SchymanskiJuliane HollenderHeinz Singer&Thank you for your attention! ! !Special thanks to…This research has been co-financed by the European Union and Greek national funds through the Operational Program "Education and Lifelong Learning" of the National Strategic Reference Framework (NSRF) ARISTEIA 624 (TREMEPOL project).

Target screening of 2327 emerging pollutants in environmental samples by RP-LC-QTOF-HR-MS/MS with an accurate-mass database

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Target screening of 2327 emerging pollutants in environmental samples by RP-LC-QTOF-HR-MS/MS with an accurate-mass database

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Digital Repository of Hellenic Managing Authority of the Operational Programme "Education and Lifelong Learning" (EDULLL)

Target screening of 2327 emerging pollutants in environmental samples by RP-LC-QTOF-HR-MS/MS with an accurate-mass database

Abstract

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Full text

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Digital Repository of Hellenic Managing Authority of the Operational Programme &quot;Education and Lifelong Learning&quot; (EDULLL)

Digital Repository of Hellenic Managing Authority of the Operational Programme "Education and Lifelong Learning" (EDULLL)