Pesticides Screening in Drinking Water by 2D LC/QToF
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1 Pesticides Screening in Drinking Water by 2D LC/QToF Jacob Samuel, Boston University School of Medicine, Biomedical Forensic Sciences Sabra Botch-Jones, Boston University School of Medicine, Biomedical Forensic Sciences Claude Mallet, Workflow Integration Group, Waters Corporation
2 Enrichment Extraction Protocol Class A (Circa 199) Traditional During Graduate Years Enrichment Extraction Protocol Class A (Circa 213) During Industrial Years 1:1 enrichment from sample protocol 1 of final extract use for analysis
3 The dreadful Extraction Procedure Prepare Sample SPE Barrels Condition Cartridge What if? 1 ml Load Sample 3 hrs drinking water 15 hrs surface water A Wash 1, 2 etc 1 ml Elute IPA, MeOH, ACN etc SPE Extraction Manifold 1. ml (1:1).1 ml (1 ) Solvent Conversion Inject 1-2 hrs N 2 Evaporation Initial LC condition - RP 95 Aqueous/5 organic B C
4 Macro vs Micro Extraction Protocol Macro Extraction Protocol min Condition 5 ml ACN/Dichloromethane (1:1) Condition 5 ml Methanol 6 min Condition 5 ml Water Micro Extraction Protocol min Condition 52 ml ACN/Dichloromethane (1:1) 6 min Condition 52 ml Methanol Condition 52 ml Water 12 min 6 min Load Sample ml/min Milli Q: 2-3 hrs Tap Water: 5-6 hrs Surface Water: hrs Wash 5 ml Water Air Dry 1 min Elute 35 ml ACN/Dichloromethane (1:1) Evaporate to dryness.1 ml (N, 35 2 C, ~3min ) Reconstitute 1 ml ACN/Water (1:1) (1:9) 16 min Load Sample 2 ml up to 2 ml Wash2 ml water Elute 1mL ACN Inject from 1 to 1 L 3 min 2:1 enrichment from sample protocol 1:1 enrichment from injection volume 2:1 final enrichment Up to 1 of final extract use for analysis Inject 1 L 3. to 15 hrs 1:1 enrichment from sample protocol 1 of final extract use for analysis
5 2D LC AT Column Dilution Hardware waste ACD Mixer D1 D2 MS Injector Loop A B Bin Pump Loader & Dilutor Bin Pump Elution
6 1D vs 2D Workflow Parameters D LC 1: MRM of 2 Channels ES+ TIC (Amphetamine) 3.11e5 1 µl MeOH 1 2D LC : MRM of 2 Channels ES+ TIC (Amphetamine) 1.56e6 1 µl ACN : MRM of 2 Channels ES TIC (Amphetamine) 7.2e4 1 µl 5/5 Water/MeOH : MRM of 2 Channels ES TIC (Amphetamine) 1.42e6 1 µl MeOH : MRM of 2 Channels ES TIC (Amphetamine) 2.56e4 1 µl Water : MRM of 2 Channels ES TIC (Amphetamine) 1.67e6 1 µl Water
7 What s next? How to optimize? OP s Carbamate Phenyl Urea Triazole Triazine Azinphos-methyl Azinphos-ethyl Mevinphos Parathion Parathion-methyl Propetamphos Dichlorovos Dimethoate Malathion Diazinon Fenitrothion Dicrotophos Chlorpyrifos 13 Others Carbofuran Carbofuran 3-keto Carboruran 3-hydroxy Methiocarb Prosulfocarb Cabetamide Aminocarb Carbaryl Fenubucarb Oxamyl Aldicarb Aldicarb-Sulfoxide Aldicarb-Sulfone Fenoxycarb 14 Diuron Linuron Monolinuron Monuron Chlortoluron Isoproturon Metoxuron Metsulfuron-Methyl Thifensulfuron-methyl Buturon Chlorbromuron Chloroxuron Difenoxuron Dimefuron Fenuron Fluometuron Metobromuron Siduron Thidiazuron 2 Carbendazim Cyproconazole Propiconazole Tebuconazole Difenoconazole Ketoconazole Itraconazole Voriconazole Posaconazole Ravuconazole 1 Atrazine Atrazine Desethyl Atrazine Desisopropyl Atrazine Desisopropyl 2- hydroxy Atrazine Desethyl desisopropyl Atrazine Desethyl Desisopropyl 2- hydroxy Propazine Simazine Terbutryn Trietazine Ametryn Prometryn Terbuthylazine Simetryn 14 Bromoxynil Propyzamide Diflufenican Flufenacet Chloridazon Flusilazole Asulam Bentazon Florasulam Pendimethalin Total of 91 1
8 Triazines Tri Organophosphorous Phenyl Urea Triazoles Carbamates Wide selection of chemical and physical properties
9 2D LC &Trap Column - Hardware 2D UPLC 2.1 x 2 mm 2.1 x 3 mm Trap Column > 1 µm particle 2. µm Fritz Analytical Column 2.1 x 5 mm UPLC 1.7 µm particle.2 µm Fritz.1 1 ml/min flow rate
10 2D Workflow Method Optimization Loading ph Condition Loading Chemistries Eluting Chemistries Eluting Organic & ph Condition ph 3 (2 FA) ph 7 (no additive) ph 1 (2 NH 4 OH) Polymer (2 µm) (high retention) Si-C 18 (1 µm) (intermediate retention) Si-C 8 (1 u m) (low retention) C 18 (1.7 um) C 8 (1.7 um) HSS T3 Low ph mobile phase (.5 FA, ph 3.2) Hi ph mobile phase (.5 NH 4 OH, ph 1) Ion Pair mobile phase 2 mm NH 4 Formate ph mm NH 4 Acetate ph mm NH 4 Bicarbonate ph 1 Elution Strength Methanol, Acetonitrile, Acetone > 4 possible permutations: Start with top 36 methods
11 Fig 4 6x6 Method Optimization (36 permutations) 1 min LC Run / 18 hours 3x3 Method Optimization (9 permutations) 1 min LC Run / 4.5 hours 3x6 Method Optimization (18 permutations) 1 min LC Run / 9 hours Loading ph 7 C8 ph 1 Loading ph 3 ACN Elution C8 4 ph 7 Loading ph 3 ACN Elution C8 1 ph 3 Loading ph 3 ACN Elution 8 C18 ph 1 Loading ph 3 ACN Elution C18 5 ph 7 Loading ph 3 ACN Elution C18 2 ph 3 Loading ph 3 ACN Elution 9 HLB ph 1 Loading ph 3 ACN Elution HLB 6 ph 7 Loading ph 3 ACN Elution 3 HLB ph 3 Loading ph 3 ACN Elution Trap Retention Strength Loading ph C8 16 ph 1 Loading ph 1 ACN Elution C8 13 ph 7 Loading ph 1 ACN Elution C8 1 ph 3 Loading ph 1 ACN Elution C18 17 ph 1 Loading ph 1 ACN Elution C18 14 ph 7 Loading ph 1 ACN Elution C18 11 ph 3 Loading ph 1 ACN Elution Elution ph HLB 18 ph 1 Loading ph 1 ACN Elution HLB 15 ph 7 Loading ph 1 ACN Elution HLB 12 ph 3 Loading ph 1ACN Elution Loading ph 25 C8 ph 1 Loading ph 3 MeOH Elution C8 22 ph 7 Loading ph 3 MeOH Elution 19 C8 ph 3 Loading ph 3 MeOH Elution 26 C18 ph 1 Loading ph 3 MeOH Elution C18 23 ph 7 Loading ph 3 MeOH Elution C18 2 ph 3 Loading ph 3 MeOH Elution 27 HLB ph 1 Loading ph 3 MeOH Elution HLB 24 ph 7 Loading ph 3 MeOH Elution 21 HLB ph 3 Loading ph 3 MeOH Elution Elution Polarity 34 C8 ph 1 Loading ph 1 MeOH Elution C8 31 ph 7 Loading ph 1 MeOH Elution C8 28 ph 3 Loading ph 1 MeOH Elution 35 C18 ph 1 Loading ph 1 MeOH Elution C18 32 ph 7 Loading ph 1 meoh Elution C18 29 ph 3 Loading ph 1 MeOH Elution 36 HLB ph 1 Loading ph 1 MeOH Elution HLB 33 ph 7 Loading ph 1 MeOH Elution HLB 3 ph 3 Loading ph 1 MeOH Elution
12 2D Workflow Method Optimization Analyte vs 36 methods Microcystins Azinphos-methyl(Water) e5 e6 e5 e5 e5 e4 e5 e5 e5 Azinphos-methyl(ACN) e6 e6 e5 e5 e5 e5 e6 e6 e5 Azinphos-methyl(MeOH) e6 e6 e5 e5 e5 e5 e6 e6 e Azinphos-methyl(Water) e5 e5 e5 e5 e5 e5 e5 e5 e5 Azinphos-methyl(ACN) e6 e6 e5 e5 e5 e5 e6 e5 e5 Azinphos-methyl(MeOH) e6 e6 e5 e6 e5 e5 e6 e6 e Azinphos-methyl(Water) e5 e5 tail e5 e5 blob e5 e5 tail Azinphos-methyl(ACN) e5 e6 tail e5 e6 blob e5 e6 tail Azinphos-methyl(MeOH) e5 e6 tail e5 e6 blob e5 e6 tail Azinphos-methyl(Water) e5 e5 tail e5 e5 tail e5 e5 RB Azinphos-methyl(ACN) e6 e6 tail e5 e6 tail e5 e6 tail Azinphos-methyl(MeOH) e6 e6 tail e5 e6 tail e5 e6 tail Meth 7 H 2 O spike 1 1: MRM of 2 Channels ES+ TIC (Azinphos Methyl) 1.2e6 Meth 1 MeOH spike 1 1: MRM of 2 Channels ES+ TIC (Azinphos Methyl) 1.2e Meth 21 ACN spike 1 1: MRM of 2 Channels ES+ TIC (Azinphos Methyl) 1.2e
13 2D Workflow Method Optimization Classes vs 36 methods Organophosphorous Azinphos-methyl(Water) e5 e6 e5 e5 e5 e4 e5 e5 e5 Azinphos-methyl(ACN) e6 e6 e5 e5 e5 e5 e6 e6 e5 Azinphos-methyl(MeOH) e6 e6 e5 e5 e5 e5 e6 e6 e5 Carbarmate Carbofuran(Water) e8 e8 e7 e8 e8 e8 e8 e8 e8 Carbofuran(ACN) e8 e8 e7 e8 e8 e8 e8 e8 e8 Carbofuran(MeOH) e8 e8 e8 e8 e8 e8 e8 e8 e8 Phenyl Urea Diuron(Water) e6 e6 e6 e7 e6 e7 e6 e7 e7 Diuron(ACN) e6 e6 e6 e7 e7 e7 e7 e7 e7 Diuron(MeOH) e6 e6 e6 e7 e7 e7 e7 e7 e7 Triazole Carbendazim(Water) e5 e6 e5 e6 e5 e4 e6 Carbendazim(ACN) e5 e5 e6 e6 e6 Carbendazim(MeOH) e5 e6 e6 e6 e4 e4 e6 Triazine Atrazine(Water) e6 e6 e6 e8 e8 e8 e7 e8 e7 Atrazine(ACN) e6 e6 e6 e8 e8 e8 e7 e8 e7 Atrazine(MeOH) e6 e6 e6 e8 e8 e8 e7 e8 e7 Others Bromoxynil(Water) e4 MP split e4 MP Bromoxynil(ACN) e4 MP split e4 MP Bromoxynil(MeOH) e4 MP split e4 MP
14 2D Workflow Method Optimization One Class vs 36 methods Azinphos-Methyl Azinphos-Ethyl Mevinphos Parathion Parathion-Methyl Propetamphos Dichlorvos ACN ph 3 MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN MeOH ACN 1- C8 ph 3 e6 e6 e6 e6 e6 e7 e6 e6 e5 e5 e6 e6 e6 e6 e6 e6 e7 e7 e8 e8 e6 e6 e7 e8 e7 e C18 ph 3 e6 e6 e6 e6 e6 e6 e6 e6 e5 e5 e6 e6 e6 e6 e7 e7 e7 e7 e8 e8 e5 e5 e7 e8 e7 e HLB ph3 e5 e5 e5 e6 e7 e6 e6 e5 e5 e5 e5 e6 e6 e6 e7 e7 e7 e7 e8 e8 e5 e5 e7 e6 e C8 ph 7 e5 e5 e6 e6 e6 e7 e6 e6 e6 e5 e6 e6 e6 e6 e6 e6 e7 e8 e8 e8 e6 e6 e7 e8 e7 e C18 ph 7 e5 e5 e6 e6 e6 e6 e6 e6 e5 e6 e6 e6 e7 e7 e7 e7 e7 e8 e8 e6 e6 e8 e8 e8 e7 e HLB ph 7 e5 e5 e5 e5 e6 e7 e6 e6 e5 e5 e5 e5 e6 e6 e6 e7 e7 e7 e8 e8 e5 e5 e7 e7 e6 e C8 ph 1 e6 e6 e6 e6 e6 e7 e6 e6 e5 e5 e6 e6 e6 e6 e7 e6 e8 e8 e8 e8 e6 e6 e7 e8 e7 e C18 ph 1 e6 e6 e6 e6 e6 e6 e6 e6 e5 e5 e6 e6 e6 e6 e7 e7 e8 e8 e8 e8 e6 e6 e7 e8 e7 e HLB ph 1 e5 e5 e5 e5 e6 e7 e6 e6 e5 e5 e5 e5 e6 e6 e7 e7 e7 e7 e8 e8 e5 e5 e7 e7 e7 e7 13 ACN ph 1 1- C8 ph 3 e6 e6 e6 e6 e7 e7 e6 e6 e5 e5 e6 e6 e6 e6 e6 e6 e8 e8 e8 e8 e5 e5 e7 e7 e7 e C18 ph 3 e6 e6 e6 e6 e7 e7 e6 e6 e5 e5 e6 e6 e6 e6 e7 e7 e8 e8 e8 e8 e5 e5 e8 e8 e7 e HLB ph 3 e5 e5 e6 e6 e7 e7 e5 e5 e4 e4 e6 e6 e6 e6 e7 e7 e7 e7 e8 e8 e4 e5 e8 e8 e7 e C8 ph 7 e5 e6 e6 e6 e7 e7 e6 e6 e4 e4 e6 e6 e6 e6 e7 e7 e8 e8 e8 e8 e5 e5 e7 e8 e7 e C18 ph 7 e5 e5 e6 e6 e7 e7 e6 e6 e5 e5 e6 e6 e6 e7 e7 e7 e8 e8 e8 e8 e5 e5 e7 e8 e7 e HLB ph 7 e5 e5 e5 e5 e7 e7 e5 e5 e4 e4 e6 e6 e6 e6 e7 e7 e7 e7 e8 e8 e4 e5 e7 e8 e7 e C8 ph 1 e6 e6 e6 e6 e6 e7 e6 e6 e4 e4 e6 e6 e6 e6 e6 e7 e8 e8 e8 e8 e5 e5 e7 e7 e7 e C18 ph 1 e5 e6 e6 e6 e7 e7 e6 e6 e4 e5 e6 e6 e5 e6 e7 e7 e8 e8 e8 e8 e5 e5 e7 e8 e7 e HLB ph 1 e5 e5 e5 e5 e6 e7 e6 e6 e4 e4 e6 e6 e6 e6 e7 e7 e7 e7 e8 e8 e5 e7 e8 e7 e7 12 MeOH ph C8 ph 3 e5 e5 e5 e5 e6 e6 e7 e7 e6 e6 e6 e6 e7 e7 e6 e6 e7 e7 e8 e8 e6 e6 e7 e7 e7 e C18 ph 3 e6 e6 e6 e6 e7 e7 e6 e6 e6 e6 e7 e7 e6 e8 e7 e8 e8 e6 e6 e7 e7 e8 e HLB ph3 e5 e5 e6 e6 e C8 ph 7 e5 e5 e5 e5 e6 e6 e7 e7 e6 e6 e5 e5 e7 e7 e6 e6 e7 e7 e8 e8 e6 e6 e7 e7 e7 e C18 ph 7 e6 e6 e6 e6 e6 e7 e7 e7 e6 e6 e6 e6 e7 e7 e7 e7 e7 e8 e8 e6 e6 e7 e7 e8 e HLB ph 7 e7 e7 e7 e C8 ph 1 e5 e5 e5 e5 e6 e6 e7 e6 e6 e6 e5 e5 e7 e7 e6 e7 e7 e8 e8 e6 e6 e7 e7 e7 e C18 ph 1 e6 e6 e6 e6 e6 e7 e7 e7 e6 e6 e6 e6 e7 e7 e7 e7 e8 e8 e8 e6 e6 e7 e7 e8 e HLB ph 1 e7 e7 e7 2 MeOH ph C8 ph 3 e6 e6 e5 e5 e6 e7 e6 e6 e5 e5 e6 e6 e7 e7 e6 e6 e7 e7 e8 e8 e6 e6 e8 e7 e7 e C18 ph 3 e6 e6 e6 e6 e6 e7 e7 e7 e5 e5 e6 e6 e7 e7 e6 e7 e7 e7 e8 e8 e6 e6 e7 e7 e7 e HLB ph C8 ph 7 e5 e5 e5 e5 e6 e7 e6 e6 e5 e5 e6 e6 e7 e7 e6 e6 e7 e7 e8 e8 e5 e5 e7 e7 e7 e C18 ph 7 e6 e6 e6 e6 e7 e7 e6 e6 e5 e5 e6 e6 e7 e7 e7 e7 e7 e8 e8 e6 e6 e7 e7 e7 e HLB ph C8 ph 1 e5 e5 e5 e5 e6 e7 e6 e6 e5 e5 e5 e5 e7 e7 e7 e6 e7 e7 e7 e7 e5 e5 e7 e7 e7 e C18 ph 1 e6 e6 e6 e6 e6 e7 e6 e6 e5 e5 e6 e6 e7 e7 e7 e5 e6 e8 e8 e6 e6 e7 e7 e7 e HLB ph 1 Dimethoate Malathion Diazinon Fenitrothion Dicrotophos Chlorpyrifos
15 2D Workflow Method Optimization Top 4 methods Total Coverage Group 1 Method 9 Group 2 - Method 5 (14) & 9 (12) Group 3 - Method 14 & 5 Group 4 - Method 14 Group 5 - Method 14 Group 6 - Method 12 (9) Green box: Blue box: Methods: highest signal 1/1 lower Group 1 (13) Group 2 (14) ACN ph 3 1- C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph ACN ph 1 1- C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph MeOH ph C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph MeOH ph C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph C8 ph C18 ph HLB ph Group 3 (2) Group 4 (12) Group 5 (14) Group 6 (1)
16 2D Workflow Instrument LOD QQQ (MRM s) Group 1 LOD Chlorpyrifos Diazinon Dichlorvos 5 ppt Dicrotophos Dimethoate Fenitrothion 2 ppt Malathion 2 ppt Mevinphhos 5 ppt Parathion Parathion Methyl Propetamphos 5 ppt Group 4 LOD Fluconazole Tebuconazole Propiconazole Voriconazole Difenoconazole Ravuconazole Ketoconazole Posaconazole Itraconazole 5 ppt Group 2 Carbaryl Fenobucarb Aminocarb Aldicarb Carbofuran Aldicarb Sulfone Methiocarb Carbetamide Carbofuran 3 OH Oxamyl Prosulfocarb Fenoxycarb Group 5 Prometryn Terbutryn Propazine Trietazine Terbuthylazine Ametryn Simetryn Simazine Atrazine Desethyl Atrazine Desisopropyl LOD 5 ppt 2 ppt 5 ppt 5 ppt LOD 5 ppt 5 ppt Group 3 Monuron Thidiazuron Fluometuron Siduron Metobromuron Difenoxurone Chloroxuron Chlorobromuron Dimefuron Tribenuron methyl Fenuron Isoproturon Chlortoluron Monolinuron Metoxuron Diuron Buturon Linuron Metsulfuron Methyl Group 6 Chloridazon Asulam Bentazon Propysamide Pendimethallin Flusilazole Florasulam Flufenacet Diflufenican LOD 2 ppt 5 ppt 2 ppt 5 ppt LOD 2 ppt 2 ppt 2 ppt
17 2D Detection Workflow Limits Instrument LOD QToF (Scan) Group 1 Chlorpyrifos Malathion Diazinon Parathion Propetamphos Fenitrothion Parathion Methyl Dicrotophos Dimethoate Mevinphos Dichlorvos Group 4 Fluconazole Tebuconazole Propiconazole Voriconazole Difenoconazole Ravuconazole Ketoconazole Posaconazole Itraconazole LOD LOD Group 2 Carbaryl Fenobucarb Aminocarb Aldicarb Carbofuran Aldicarb Sulfone Methiocarb Carbetamide Carbofuran 3 OH Oxamyl Prosulfocarb Fenoxycarb LOD Group 5 LOD Prometryn Terbutryn Propazine Trietazine Terbuthylazine Ametryn Simetryn Simazine Atrazine Desethyl Atrazine Desisopropyl Group 3 Monuron Thidiazuron Fluometuron Siduron Metobromuron Difenoxurone Chloroxuron Chlorobromuron Dimefuron Tribenuron methyl Fenuron Isoproturon Chlortoluron Monolinuron Metoxuron Diuron Buturon Linuron Metsulfuron Methyl Group 6 Chloridazon Asulam Bentazon Propyzamide Pendimethallin Flusilazole Florasulam Flufenacet Diflufenican LOD LOD
18 2D Workflow - LOD QQQ TIC (Diazinon) e8 1 pg/ml 6.42 TIC (Propetamphos) 1.2e TIC (Diazinon) e TIC (Diazinon) e5 1 pg/ml 6.73 TIC (Diazinon) 6.61e TIC (Propetamphos) e TIC (Propetamphos) e TIC (Propetamphos) e4 1 pg/ml 1 pg/ml
19 2D Workflow - LOD QToF 1 1 ng/ml 4.1 Florasulam 2.35e Bentazon 6.18e Florasulam 5.6e Bentazon e Florasulam 8.49e Bentazon e3 1 ng/ml 1 ng/ml
20 2D Workflow RP Extraction Few Facts 1-1 ml drinking 1 ml/min flow rate 2-1 minutes loading minimum risk of clogging 3- quick wash step 4- Elute with 2 ml final 5:1 enrichment 5- inject 25 ul if more sensitivity is needed: Option 1: 2 ml loading at 2 min for 1:1 enrichment Option 2: Inject 5 ul
21 2D Workflow Extraction Strategy Wash: 1 ph 1 Basic Functionality (green) - High organic elution at high ph - low organic elution at low ph Polarity/solubility (yellow) OP Dichlorvos ACN ph ACN ph MeOH ph MeOH ph Mevinphos ACN ph ACN ph MeOH ph MeOH ph Dimethoate ACN ph ACN ph MeOH ph MeOH ph Dicrotophos ACN ph ACN ph MeOH ph MeOH ph Elute: 3 ph 3 Objectives: 1- Maximum Organic for wash for zero analyte breakthrough 2- Minimum Organic for total elution and total coverage
22 2D Workflow Extraction Strategy Red box Minimum for Elution Blue Box Maximum for Wash OP Dichlorvos ACN ph ACN ph MeOH ph MeOH ph Mevinphos ACN ph ACN ph MeOH ph MeOH ph Dimethoate ACN ph ACN ph MeOH ph MeOH ph Dicrotophos ACN ph ACN ph MeOH ph MeOH ph Parathion Early ACN ph1 Eluters ACN ph MeOH ph MeOH ph Fenitrothion ACN ph ACN ph MeOH ph MeOH ph Propetamphos ACN ph ACN ph MeOH ph MeOH ph Late Eluters
23 2D Workflow Extraction Strategy RP W1 (low ph) W2 (high ph) Elution Organophosphorous 1 MeOH 2 MeOH 1 MeOH low ph Carbamates x 2 MeOH 1 MeOH low ph Triazole x 2 MeOH 1 MeOH low ph Pheny urea (A) 4 MeOH 1 MeOH 1 MeOH low ph Phenyl urea (B) 2 MeOH x 1 MeOH low ph Others 1 MeOH x 9 MeOH low ph Triazines 2 MeOH 2 MeOH 9 MeOH low ph Six distinct extraction protocol for each classes
24 RP Extraction QQQ 5 ppt ) Close to saturation point suggesting lower detect limit is possible 1 Stable baseline 6.19 TIC (Prometryn) 2.68e TIC (Ametryn) 2.13e TIC (Terbutryn) 2.68e TIC (Atrazine) 9.71e TIC (Propazine) 2.59e TIC (Trietazine) 1.82e TIC (Simetryn) 2.48e TIC (Simazine) 2.4e TIC (Terbuthylazine) 1.84e TIC (Atrazine Desisopropyl) 2.21e
25 RP Extraction QQQ (OP 5 ppt) Close to background noise reaching lower detect limit would be difficult TIC (Chlorpyrifos) 3.4e TIC (Propetamphos) 3.65e TIC (Malathion) 1.45e TIC (Diazinon) 2.68e TIC (Parathion) 2.8e TIC (Dimethoate) 4.33e Stable & clean baseline 6.35 TIC (Fenitrothion) 3.63e TIC (Parathion Methyl) 4.96e TIC (Dicrotophos) 2.2e TIC (Dichlorvos) 1.75e
26 RP Extraction TOF ) e6 Prometryn e6 Amteryn e6 Terbutryn e4 Atrazine e Simetryn 1.18e5 Propazine Trietazine e e Terbuthylazine Atrazine Desisiopropyl Simazine e e
27 2D Workflow RP/IE Extraction Step 1: Load 2 ml Step 2: disconnect barrel RP / Anion Exchange RP/Anion RP/Cation Wash 1: 1 H 2 O + 2 NH 4 OH Elute 1: 1 MeOH + NH 4 OH Elute 2: 1 MeOH + 2 FA Wash 1: 1 H 2 O + 2 FA Elute 1: 1 MeOH + 2 FA Elute 2: 1 MeOH + NH 4 OH RP / Cation Exchange Elute 1: Neutral and Basic moities Elute 2: Acidics moities Elute 1: Neutral and Acidic moities Elute 2: Basic moities One Protocol two fractions Basic behavior Zwitter behavior MAX MAX MCX MCX Elute 1 Elute 2 Elute 1 Elute 2 Dichlorvos Mevinphos Dimethoate Dicrotophos Parathion Methyl Fenitrothion Propetamphos Parathion Diazinon Chlorpyrifos Malathion
28 2D Workflow RP/IE Extraction MAX MAX MCX MCX Others Elute 1 Elute 2 Elute 1 Elute 2 Chloridazon Asulam Propyzamide Bentazon Pendimethallin Flusilazole Florasulam Flufenacet Diflufenican Acidic behavior MAX MAX MCX MCX Triazines Elute 1 Elute 2 Elute 1 Elute 2 Prometryn Terbutryn Propazine Trietazine Terbuthylazine Ametryn Atrazine Simetryn Simazine Atrazine Desethyl Atrazine Desisopropyl MAX MAX MCX MCX Phenyl Urea Elute 1 Elute 2 Elute 1 Elute 2 Tribenuron Methyl Dimefuron Chlorobromuron Chloroxuron Difenoxuron Metobromuron Siduron Fluometuron Thidiazuron Monuron Fenuron Isoproturon Chlortoluron Monolinuron Metoxuron Diruon Buturon Linuron Metsulfuron Methyl Basic behavior Breakthrough behavior 6 mg SPE bed not enough
29 MAX/MCX Extraction QQQ (Triazines at 5 ppt) Elute 1 on MAX Close to saturation point suggesting lower detect limit is possible TIC (Prometryn) 2.68e TIC (Ametryn) 2.68e TIC (Terbutryn) 2.68e TIC (Atrazine) 1.43e TIC (Propazine) 2.68e TIC (Simetryn) 2.68e TIC (Trietazine) 2.68e TIC (Terbuthylazine) e TIC (Simazine) 4.49e TIC (Atrazine Desisopropyl) 3.77e
30 MAX/MCX Extraction QQQ (OP 5 ppt) Elute 1 on MAX Higher signal than RP protocol reaching lower detect limit would be possible signal increase TIC (Chlorpyrifos) 8.48e TIC (Malathion) 8.65e signal increase 6.41 TIC (Propetamphos) 3.73e6 TIC (Fenitrothion) 6.57e TIC (Diazinon) 2.68e8 1 5 signal increase TIC (Parathion Methyl) 1.38e TIC (Parathion) 3.44e TIC (Dimethoate) e TIC (Dicrotophos) 2.24e TIC (Dichlorvos) 2.1e
31 MCX/MAX Extraction TOF ) 1 Prometryn e e6 Amteryn e Terbutryn e5 Atrazine e6 1.56e Propazine Trietazine Terbuthylazine e e Atrazine Desisiopropyl 5.16 Simetryn Simazine e e
32 Conclusions - Fast sample preparation protocol > 3 min - 1:1 enrichment factor - No Evaporation to Dryness or Reconstitution - No evaporative loss - Large volume injection 25 ul injection - Sub ppb detection limits with ToF
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