Development of an extraction and clean-up method for suspect and non-target screening of emerging organic pollutants in biota using LC-HRMS
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1 Development of an extraction and clean-up method for suspect and non-target screening of emerging organic pollutants in biota using LC-HRMS WIEBKE DÜRIG 1 ; KARIN WIBERG 1 ; LUTZ AHRENS 1 1 Dept. of Aquatic Sciences and Assessment, Swedish University of Agricultural Sciences, Box 7050, SE Uppsala, Sweden, wiebke.durig@slu.se
2 Aims/Objectives Extraction and clean-up technique for nontarget purpose in biota Extraction and clean-up suitable for a broad range of compounds Spiked white-tailed sea eagle muscle tissue for evaluation with 277 compounds Evaluation by matrix effect and recovery
3 Modified from Krauss et al. (2010) Anal. Bioanal. Chem 397: Lutz Ahrens Sampling, extraction, clean-up, fractionation LC/GC separation and HR-MS analysis Target analysis Suspect Screening Compound library Non-target screening Exact mass filtering Molecular formula (MS isotope/adducts) Structure (Database search, MS/MS) Candidate selection (RT, MS/MS) Confirmation and quantification (MS/MS, RT, reference standards )
4 Sample preparation - Biota Homogenization Drying sample Extraction Non-destructive Non-selective (broad range of compounds) Fractionation Filtration Centrifugation Sulphuric acid Basic treatment Non-destructive Clean-up Non-selective (broad range of compounds) But still reduce interference (lipids, proteins, etc.) Fractionation Instrumental analysis
5 Sonication vs QuECHERS 1g White-tailed sea eagle muscle tissue Homogenization with 5mL ACN Sonication (30 minutes) QuECHERS 900 mg MgSO4 300 mg Z-Sep+ Freezer for >16 hours (denaturalize proteins)
6 Clean-up SPE (PRiME HLB) PRiME HLB (Waters) Process Robustness Improvements in Matrix effects Ease of use Reversed-phase SPE 1 ml extract + 4 ml Milli Q PRiME HLB Inject extract on cartidges Wash catridges with 5 ml of 5% MeOH Elute with 5 ml of 100% MeOH
7 Evaluation of extraction techniques Matrix effect 1g White-tailed sea eagle muscle tissue spiked 200ng native + IS Homogenization with 5mL ACN Sonication (30 minutes) QuECHERS 900 mg MgSO4 300 mg Z-Sep+ Freezer for >16 hours (denaturalize proteins) SPE SPE Solvent blank Matrix Effect > 1 -> Matrix enhancement Matrix Effect < 1 -> Matrix supression spiked 200ng native + IS spiked 200ng native + IS Final extract Final extract
8 Evaluation of extraction techniques spiked 200ng native + IS Recovery 1g White-tailed sea eagle muscle tissue spiked 200ng native + IS Solvent blank = 100 % Homogenization with 5mL ACN Sonication (30 minutes) QuECHERS 900 mg MgSO4 300 mg Z-Sep+ Freezer for >16 hours (denaturalize proteins) SPE SPE Final extract Final extract
9 Results Compounds (n= 277) Log K ow : -2,00 till 10,43 pos [PERCE NTAGE] n=196 neg [PERCE NTAGE] n=81 ESI + ESI - Molecular weight: 119,127 till 837,047 Benzotriazoles/ Benzothiazoles (n= 7) Contrast Media (n=1) Drugs (n=4) Endocrine Disrupting Compounds (n=4) Detected compounds Food Additives (n=4) Flame retardants (n=17) Personal Care Products (n=5) Pesticides (n=90) PFAS (n=14) Pharmaceuticals (n=77) Phthalates (n=3)
10 QuECHERS + SPE: Matrix effect for all compounds Average matrix effect: 0,90 Average matrix effect: 0,90
11 Sonication + SPE: Matrix effect for all compounds Average matrix effect: 0,91 Average matrix effect: 0,89
12 Matrix effect for each compound class QuECHERS Sonication
13 QuECHERS + SPE: % Recovery for all compounds Average recovery: 7% Average recovery: 13%
14 Sonication + SPE: % Recovery for all compounds Average recovery: 28% Average recovery: 27%
15 Conclusion Matrix effect: low for both extraction methods Recovery: poor, but better for sonication extraction than for QuECHERS Future: of recovery by another clean-up techniqimprovementsue, like GPC Investigate losses
16 Acknowledgement Associated Professor Lutz Ahrens Professor Karin Wiberg Professor Anders Bignert Professor Peter Haglund Dr. Pablo Gago-Ferrero Samples were provided by the Natural history museum Stockholm (Naturhistorisk riksmusset)
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