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14 SPE LC GC LC/G C MS : / (LLE) (SLE) SPE : SPE (PPT) LLE LLE SPE SPE : SPE SPE / ph, pk, log P 13

15 SPE SPE SOLA HRP,C18,C8 Phenyl Retain PEP SOLA HRP,C18,C8 Retain PEP SOLA HRP,Hypercard Retain PEP SCX SAX Aminopropyl & SOLA SCX,Verify CX Retain CX & SOLA SAX,Verify AX Retain AX SOLA HRP,C18,C8, Phenyl Retain PEP Silica,Aminopropyl Cyano, Diol Silica,Florisil,Aminopropyl Cyano, Diol SCX SAX Aminopropyl & Verify CX & Verify CX 14

16 Thermo SPE ( ) - HyperSep Retain PEP/SOLA HRP m 2 /g 40 60µm 55 90Å PH 1-14 C18 ( 3-5 ) HyperSep Retain-CX/SOLA SCX m 2 /g 40 60µm 55 90Å HyperSep Retain-AX/SOLA SAX m 2 /g 40 60µm 55 90Å - HyperSep C m 2 /g 40 60µm 60Å HyperSep C8 C m 2 /g 40 60µm 60Å C8 C18 HyperSep Phenyl m 2 /g 40 60µm 60Å Phenyl HyperSep Hypercarb 100% ph SPE 15

17 - - HyperSep Silica 530m 2 /g 40 60µm 60Å HyperSep Florisil 289m 2 /g 40 60µm 60Å Florisil AOAC EPA (PCB) HyperSep Cyano m 2 /g 40 60µm 60Å Cyano HyperSep Diol m 2 /g 40 60µm 60Å HyperSep Aminopropyl m 2 /g 40 60µm 60Å HyperSep SAX m 2 /g 40 60µm 60Å SAX HyperSep SCX m 2 /g 40 60µm 60Å SCX HyperSep Verify-CX m 2 /g 40 60µm 60Å Verify-CX C8 HyperSep Verify-AX m 2 /g 40 60µm 60Å Verify-AX C8 THC 16

18 SPE mm mm SPE PH SPE ph SPE SPE / SPE 1mL/min

19 SPE SPE SPE Silica,Florisil / C18 C8 PEP SOLA HRP / 1-2ml/min 1-2ml/min / 5:95 100% (Retain CX/SOLA SCX/Verify CX) / (Retain AX/SOLA SAX/Verify AX) / 1-2ml/min 1-2ml/min 2% 100% 2% 100% 2% 2% 18

20 SPE SPE SPE SPE HyperSep SPE ph 100% : ph - ph - ph - ph ph ph ph - pka 2 ph ) - pka 2 ph ph ph ph pka pka2ph pka2ph 19

21 2 HyperSep SPE pka 3.0 pka (1-position), 1.5 (4-position)

22 HyperSep Retain PEP 30mg 1mL SPE mL 1mL 2. 1mL 500ng/mL 3. 0% /100% 90% 10% 10% mL 100% 4 HyperSep Retain PEP / 30% /70% 1.5mL 500µL HyperSep lietain-cx 30mg 1mL mL 0.1% 1mL 0.1% 2. 1mL 0.1% 500ng/mL 3. 1mL 0.1% mL 0.1% % / 20% /80% 80% /20% 10% mL 5% 4 HyperSep lietain-cx / 50% 5% /50% 5% 1.5mL 500µL HyperSep Phenyl 100mg 1mL mL 1mL ng/mL 1mL 3. 10% 90% mL 100% 4 HyperSep Phenyl / 20% /80% 1.5 ml 500µL 21

23 / / ph H ph / / / (SLE) SLE 200mL SLE LLE LLE LLE LLE : (1) SLE 5-10 (2) (3) 22

24 - SPE-LC/MS 200mg 3mL HyperSep Retain-CX ( ) 2g 0.01g 50mL 0.2mol/L ph5.2 10mL - / 40µL 3min 37 16h 3min 10min 1mol/L 1mL 10min 50mL 3mL 3mL 3mL 0.5mol/L 3mL 3mL 3mL 5% 1. LC/MS/MS LC/MS Hypersil Gold 5µm mm A: 5mM B: 1. Time(min) A(%) B(%) µL 250µL/min MS ESI SRM 4500V LOQ 0.1µg/kg 0.5 µg/kg % 2. - LC/MS/MS 1ng/mL 23

25 SPE-LC/MS 60mg 3mL HyperSep Retain-AX ( ) 5g 50mL 10mL 1min 10min 5000r/min 15mL 1mL 50 3mL 0.1mol/L NaOH ph=11.0 5min 5000r/ min 3mL 3mL 3mL 5% 3mL 5% LC/MS Hypersil Gold 5µm mm % A : B 1. Time(min) A(%) B(%) LC/MS/MS µL 250µL/min MS ESI SRM 3500V LOQ µg/kg µg/kg % 2. 6 LC/MC/MS 1ng/mL 24

26 SPE-LC/MS 500mg 3mL HyperSep C18 ( ) 5g 10% 3mL 10mL 30s 10min r/min 10min 10mL 40 30% 5mL 3mL 3mL 2mL 30% 5mL LC/MS Hypersil Gold 5µm mm % A : B 1. Time(min) A(%) B(%) µL 250µL/min MS ESI SRM 4500V

27 1. LC/MS/MS 2.11 LC/MC/MS 1.0ng/mL 2. LOQ 0.1µg/kg 0.5µg/kg % 26

28 SPE-LC/MS 500mg 3mL HyperSep Silica ( ) g 50mL 15mL 3min 10min 4000rpm 50mL 0.1mol/L NaOH 10mL 15mL mL 5mL 5mL 5mL 5mL 6:4 v/v LC/MS Hypersil Gold 5µm mm % A : B 1. Time(min) A(%) B(%) LC/MS/MS 2 LC/MC 10µL 250µL/min MS ESI SRM 3500V LOQ 0.1µg/L 0.2 µg/kg 0.2µg/L 0.2µg/kg 0.2µg/L 0.5µg/kg %

29 SPE-LC/MS 200mg 6mL HyperSep Retain-PEP ( ) 5g 50mL 100µL 1µg/mL 13 C- 20g 20mL 25g r/min 10 10mL 30s 10mL 100mL 10mL mL 4mL 6mL 6mL 2mL 5min 6mL 6mL 1~2mL/min 1. LC/MS/MS LC/MS Hypersil Gold 5µm mm A: B:0.1% 2mM 1. (min) A B (µl/min) µL 250µL/min MS ESI SRM 4500V

30 µg/kg LC/MC/MS 2. 2µg/kg 3. 50%-114% 29

31 SPE-HPLC 200mg 3mL HyperSep Retain PEP ( ) 5.00g 25mL Mcllvaine 2mL50% 6000rpmin 10min 20mL ph 4.5 3mL 3mL 3mL 5% 3mL HPLC LC/MS Hypersil Gold C18 5µm mm % A : B %B 1.5mL/min UV@350nm 25 10µL (1) (2) (3) (4) (5) 30

32 SPE-HPLC,LC/MS 200mg 6mL HyperSep Retain-AX ( ) 2g 30mL 50mM ph7.4 6mL 6mL 5M NaOH 6mL 5mL 1-2mL/min 6mL5% 1mL 6mL 4% 0.4mL HPLC Hypersil Gold aq 5µm mm A10mM KH 2 PO 4 ph3b 20%A~35%A 0~30min 1mL/min UV@280nm 35 5µL HPLC LC/MS Thermo Scientific Hypersil GOLD mm 3µm A: 0.5% B: / 0.5% 1:1 V/V 20µL 300µL/min MS ESI SRM 3000V

33 1.16 LC/MS SRM 20µg/kg % 32

34 200mg 3mL HyperSep Retain-CX ( ) 2.0g 8mL 10mmol/L KH 2 PO 4 0.4mmol/LEDTA 2% )10,000 rpm 10min 1 10,000rpm 10min 3mL 3mL 1-2mL/min 5mL 5mL 5% LC/MS LC/MS Hypersil Gold aq 5µm mm %HFBA /0.1%HFBA 52:48 20µL 200µL/min MS ESI SRM 4500V 350 (1) C1a (2) C2 33

35 SPE-LC/MS 200mg 3mL HyperSep Retain PEP ( ) 4g 8mL 2min 4000rpm 5min 5mL 4mL 1min 4000rpm 3min 1mL 50:50 / 10mL 3mL 3mL 1-2mL/min 3mL 30% 3mL µg/kg LC/MS Hypersil GDLD TM 50 mm 2.1 mm 1.9µm A: B: (min) A% % % % 20µL 500µL/min MS ESI SRM 3000V LOQ LOQ 0.050µg/kg 3. 94%-104% 34

36 SPE-LC/MS 200mg 3mL HyperSep C18 ( ) 5g 5% 5mL 0.17mol/L 5mL 50mL5000rpm 10min 3mL 3mL 1-2mL/min 3mL 6mL 20% LC/MS BioBasic AX 5µm mm A:10mM B: 10%B 20µL 200µL/min MS ESI 3000V LC/MS/MS (1) (2) (3) (4) G 35

37 SPE-HPLC 200mg 3mL HyperSep C18 ( ) 5g 20mL 2min 10000rpm 5min 20mL 5mL 20% 3mL 3mL 1-2mL/min 3mL 2*2 ml LC/MS Hypersil GOLD TM mm 5µm A: 0.1% B: + 0.1% 35:65 20µL 1mL/min UV 235nm HPLC 2. 95%-99% 36

38 SPE-HPLC 60mg 3mL HyperSep Retain PEP ( ) 15mL 25mL Mcllvaine rpm 10min 3mL 3mL 3mL 5% 3mL LC/MS Hypersil Gold 5µm mm % A : B %B 1.5mL/min UV@350nm 25 10µL HPLC (1) (2) (3) (4) (5) 37

39 SPE-LC/MS 200mg 3mL HyperSep C18 ( ) 5mL 5% 5mL 0.17mol/L 5mL 50mL5000rpm 10min 3mL 3mL 1-2mL/min 3mL 6mL 20% LC/MS/MS LC/MS BioBasic AX 5µm mm A: 10mM B: 10%B 20µL 200µL/min MS ESI 3000V 300 (1) (2) (3) (4) 38

40 60mg 3mL HyperSep Retain PEP ( ) 0.5g d5-cap 0.3 ppb 1mL 1min 14000rpm 10min 2mL 30% 3mL 3mL 1-2mL/min 3mL 30% 2mL µg/kg LC/MS Hypersil GOLD TM 50 mm 2.1 mm 1.9µm A: B: (min) A% % % % 20µL 500µL/min MS ESI SRM 3000V µg/kg 39 2.LOQ LOQ 0.050µg/kg 3. 94%-104% CAP Spiking Level (µg/kg) Within-laboratory Reproducibility n=20 Meam(%) SD(µg/kg) %RSD % % % % % % % % Table 3: Recovery and Reproducibility Data

41 60mg 3mL HyperSep Retain PEP ( ) 15mL 1 2mL 1 1mL 4,000rpm HyperSep Retain PEP 1 5mL 1 5mL 1 2mL/min 1 10mL 1 5mL 1 2mL/min 5mL <40 1,000µL 5µL LC/MS HPLC Hypersil GOLD 5µm mm SPE-LC/MS 50mg 1mL HyperSep Hypercarb ( ) 1g 12mL 50% 2min 4000rpm 10min 5mL ph=3 3mL 3mL 1-2mL/min 3mL LC/MS/MS LC/MS BioBasic TM AX mm 5µm min - -50mM 85:10:5 5min-10min 9:1 / 20µL 400µL/min MS ESI SRM 3000V

42 SPE-LC/MS 60mg 3mL HyperSep Retain-CX ( ) 5g /10mL 1% 50mL 2mL 2% 20min 3mL 3mL 6mL 2mL/min 3mL 3mL 5min 5mL 5% 1-2mL/min1mL LC/MS Syncronis HILIC 5µm mm A: B:10mM / ph=3 85:15 10µL 250µL/min MS ESI MRM 3500V 350 LC/MS/MS 41

43 SPE-LC/MS 60mg 3mL HyperSep Retain-CX ( ) 1g 2mL 5mL1% 2min 3000rpm 2min 3mL 3mL 6mL 2mL/min 3mL 3mL 5min 5mL 5% 1-2mL/min1mL LC/MS Syncranis HILIC 5µm mm A: B:10mM / ph=3 85:15 10µL 250µL/min MS ESI MRM 3500V 350 LC/MS/MS 42

44 SPE-HPLC,GC/MS 1g 6mL HyperSep Silica ( ) 2.5g 10mL 6:4 30s 5min 4000r/min 5min 40 30min-40min mL 6mL 6mL 2mL 4mL 10mL : 2:3 HPLC Hypersil Green PAH 3µm mm A: 99:1 : B: 99:1 : 50:50-5min 50:50-25min 0:100 2mL/min 10µL UV 254nm 20 PAH HPLC (1) (2) (3) (4) (5) (6) (7) (8) (9) (a) (10) (11) (b) (12) (k) (13) (a) (14) (a,h) (15) (ghi) (16) (1,2,3-cd) 43

45 GC/MS TraceGOLD TG-5SiIMS 30m 0.25mm 0.25µm TG-17SiIMS 30m 0.25mm 0.25µm min 25 /min /min 320/min 2min TraceGOLD TG-5SiIMS 90 1 min 30 /min /min 330 5min TraceGOLD TG-17Si1 MS 1.2mL/min 1µL 250 1min 30mL/min MS ppb 18 TIC 1. TG-17SiIMS 30m 0.25mm 0.25µm 10µg/mL18 TIC 2. 5SiIMS 30m 0.25mm 0.25µm 10µg/mL18 PAH TIC (a) (b) (j) (k) (a) (e) (1,2,3-cd) (1,2,3-cd) (g,h,i) 44

46 SPE-GC/MS,LC/MS QueChERS ( ) g 25mL 10mL 6g MgSO 4 1.5g Thermo QueChERS PN: min 4000r/min 2min 40 5mL g 50mL 5mL 15mL 6g MgSO 4 1.5g Thermo QueChERS PN: min 4000r/min 2min 40 5mL 50mg PSA 50mg C18 150mg MgSO 4 Thermo QueChERS clean-up PN: min 4000r/min 2min GC/MS TG-5MS 30m 0.25mm 0.25µm min 15 /min 230 1min 5 /min 280 4min 1mL/min 10µL min 50mL/min MS GC/MS 1. 25ppb

47 LC/MS Thermo AccuCore RP-MS 2.6m 100mm 2.1mm A: B: (min ) A: B: 0 60% 40% % % 0.3mL/min 3µL MS ESI SRM 3700V % 46

48 200mg 3mL HyperSep Retain-CX ( ) 1 5g 10 25mL / 50:50 5 5mL 1mL 100mM HCI 1mL 5 2mL 5 SPE HyperSep Retain-CX 1 3mL 1 3mL 3 1 2mL/min 1 3mL 100mM 1 1mL 10 5 ) 1 2mL 5%NH 4 OH 1 3mL 5%NH 4 OH 1 2mL/min 40 Syncronis HILIC 5µm mm A: B:10mM / ph=3 85:15 10µL 250µL/min MS ESI MRM 3500V 350 LC/MS/MS 47

49 LC/MS/MS 200mg 6mL HyperSep Verify-CX ( ) 200mg 6mL HyperSep Retain-AX ( ) 1 5g 10 25mL CH 3 CN/ 50:50 5 5mL 1mL 100mM HCI 1mL CH 2 CI 2 5 2mL CH 2 CI 2 5 SPE HyperSep Verify CX 1 3mL CH 3 OH 1 3mL 3 1 2mL/min HyperSep Retain-AX HyperSep Verify-CX 1 1mL HyperSep Retain-AX HyperSep Retain-AX 1 3mL100mM HCI 1 1mL CH 3 OH mL 5% NH 4 OH CH 3 OH 1 3mL 5% NH 4 OH CH 3 OH 1 2mL/min 40 1,000µL CH 3 CN * 5µL LC/MS HyperSep Retain-AX ph 7** HyperSep Retain-AX 1 3mL CH 3 OH 1 3mL 3 PH 7 1 2mL/min HyperSep Retain -AX 1 3mL 1 1mLCH 3 OH 1 3mL CH 3 OH 1% HCI 1 2mL CH 3 OH 1% HCI 1 2mL/min µL * 5 LC/MS MRM 127.1/85.1 2, 4 6- * 127.1/ /84.9 * 2, 4 6- ** µL5%(V/V)(aq)NH 4 OH PH HPLC Hypersil GOLD HILIC 3µm mm

50 SPE-LC/MS 200mg 3mL HyperSep Retain PEP ( ) 60mg 3mL HyperSep Retain-CX ( ) 2g 10mL 20min 4000rpm 10min 5mL 5min 5mL 15mL mL SPE 3mL 3mL 1-2mL/min HyperSep Retain PEP 3mL 3mL1% HyperSep Retain CX 1mL LC/MS Thermo Scientific Hypercarb mm 5µm µL 400µL/min MS ESI SRM 3500V ng/mL LC/MS 2. 1g 10% ng %~ 97.4% RSD 10% 3. Cereal 1 Cereal 2 Potato Chip 1 Potato Chip 2 Injection ng/mL 55.93ng/mL 57.11ng/mL 29.18ng/mL Injection ng/mL 56.18ng/mL 56.52ng/mL 29.14ng/mL Mean 17.09ng/mL 56.06ng/mL 56.82ng/mL 29.16ng/mL Extraction Vol. 20.0mL 20.0mL 20.0 ml 20.0mL Mass Sample 2.003g 2.007g Acrylamide Conc 171ng/g 559ng/g 562ng/g 292ng/g Table 2: Results of acrylamide assay from food samples 49

51 SPE-LC/MS 200mg 3mL HyperSep Retain-AX ( ) 2g 20mL 5% 6mL 6mL 5M NaOH 6mL 5mL 1-2mL/min 3mL 5% 3mL 3mL 4% 0.4mL % LC/MS Thermo Scientific Hypersil GOLD mm 3µm A: 0.5% B: / 0.5% 1:1 V/V 20µL 300µL/min MS ESI SRM 3000V 350 SPE-HPLC 500mg 6mL HyperSep C18 ( ) 5g 25mL 6mL 6mL 1-2mL/min 6mL 6mL 1mL LC/MS/MS 100µL N- - 1:1 30s, 150µL - 1:1 30s 750µL 15min HPLC LC/MS Hypersil Gold 5µm mm /0.1% =75:25 10µL 250µL/min MS APCI HPLC Hypersil GDLD TM 150mm 4.6mm 5µm / =92:8 20µL 1mL/min 365nm475nm 25 50

52 SPE-LC/MS 200mg 6mL HyperSep Retain-PEP ( ) 5g 15mL 20mL 100mL 20mL mL 4mL 6mL 6mL 6mL 5min 6mL 6mL 1-2mL/min 1. LC/MS/MS LC/MS Hypersil Gold 5µm mm A: B:0.1% 2mM 1. (min) A B (µl/min) µL 250µL/min MS ESI SRM 4500V

53 1.17 2µg/kg LC/MS/MS 2. 2µg/kg 3. 60%-120% 52

54 SPE-HPLC 60mg 3mL HyperSep Retain PEP ( ) 5mL 25mL Mcllvaine ph rpm 10min 3mL 3mL 3mL 5% 3mL HPLC Hypersil Gold 5µm mm % A : B %B 1.5mL/min UV@350nm 25 10µL HPLC (1) (2) (3) (4) (5) 53

55 SPE-LC/MS 200mg 3mL HyperSep SCX ( ) 200mg 3mL HyperSep C18 ( ) 10g 20mL 0.1 5min HyperSep SCX 3mL 3mL 5mL 0.1% 5mL 5mL 0.2mol/L ph=8 ph=2 HyperSep C18 HyperSep C18 3mL 3mL 5mL 5mL 1mL LC/MS LC/MS Hypersil Syncronis HILIC 5µm mm A:100mM 100mM 0.2% B:100mM A(%) B(%) Time(min) mL/min 10µL 200µL/min MS ESI SRM 4500V

56 SPE-HPLC 200mg 6mL HyperSep Retain-PEP ( ) 5g 25mL ph mol/L 6mL 6mL 2mL/min 6mL 5min 6mL 1-2mL/min1mL HPLC Betasil CN 5µm mm A: B:20mM +20mM +2mM 20µL 1.25mL/min UV 210nm 45 HPLC 55

57 SPE-LC/MS 60mg 200mg 3mL HyperSep Retain PEP ( , ) 5g 20mL 100g 60mL /1% 3:2 15mL /1% 3:2 3mL 3mL 1-2mL/min 3mL30% 3mL µg/kg LC/MS Hypersil GOLD TM 50 mm 2.1mm 1.9µm A: B: min A% % % % 20µL 500µL/min MS ESI SRM 3000V µg/kg 2.LOQ LOQ 0.1µg/kg 3. 93%-99% 56

58 60mg 3mL HyperSep Retain-CX ( ) 100µg/mL (NOR-D5) 1µg/mL 5g 50mL 50µL *MNZDMZ (HMMNI) IPZ2-(2- ) (IPZOH)RNZ 10mL 0.5mol/L 15 10mL 30 2,500rpm 3 50mL 10mL 40 10% 5mL ACN HyperSep Retain-CX 1 3mL 1 3mL 1 2mL/min 1 3mL mL 1 2mL/min 25µL LC/MS 0.20mL/min A 5mmol/L B HPLC Hypersil GOLD 3µm mm SPE-HPLC 60mg 3mL HyperSep Retain-PEP ( ) 5.00g 100mL 50µL MC-RR YR LR LW LF 1µg r/min 1min 10mL 2min 15min 5min 3000r/min 0.5 ml 10mL 3mL 3 ml 6mL 3mL LC/MS Hypersil Gold 5µm mm % A : B 0.00min 20% 80% 4.00min 95% 5% 8.00 min 95% 5% 8.10min 20% 80% min 20% 80% 10µL 250µL/min MS ESI SRM 3500V

59 1. LC/MS/MS 1.5 MCs 5µg/kg LC/MS/MS 2. MC-RR YR LR LW LF µg/kg 2. n=6 Compounds RR YR LR LW LF spiked level µg/kg Recovery % RSD % % 58

60 SPE-LC/MS 200mg 6mL HyperSep Retain-CX ( ) 2g Mcllvaines ph2.6 / =50/50 10mL 5000rmp 20 SPE 6mL 5mL 5mL Mcllvaines ph mL/min 6mL 0.1 NHCI 6mL 50% / 6mL 10mL 50% 45% 5% V/V/V LC/MS Thermo Scientific Hypersil GOLD mm 5µm A: 0.1% B: 0.1% 40%A-5%A 0-5min 20µL 300µL/min MS ESI SRM 3500V LOQ µg/kg 1. LOD µg/kg % RSD N=3 LOQ µg/kg %RSD N=3 R ng/kg ppt 50ng/kg ppt 59

61 SPE-LC/MS 60mg 3mL HyperSep Retain PEP ( ) 2g 4mL 0.5mL 0.5M HCL 200µL 50mM DMSO 1 min mL 0.1M K 2 HPO M NaOH PH mL 3mL 1-2mL/min 5mL 2mL 30% 5mL LC/MS Thermo Scientific Hypersil GOLD 5µm mm A: 0.5mM B: min %A %B µL 250µL/min MS ESI SRM 5000V

62 µg/kg 4 LC/MS 2.LOQ AHD AOZ SEM AMOZ LOQ0.05µg/kg %-110% RSD 3 22% n=3 Fortification Level (µg/kg) AHD AOZ SEM AMOZ % % 89 15% 98 14% % % % 95 6% % 86 18% 79 19% 100 3% 61

63 SPE-HPLC 200mg 6mL HyperSep Retain-PEP ( ) 5g 10mL 2min 10000rpm 5min 20mL 35 10mL ph=8 6mL 6mL 10mL 2mL/min 6mL 5min 6mL 1-2mL/min1mL LC/MS Betasil CN 5µm mm A: B: 20mM +20mM +2mM A/B 35:65 20µL 1.25mL/min UV 210nm 45 HPLC 62

64 SPE-HPLC,GC/MS 1g 6mL HyperSep Silica ( ) 6000r/min 10min 2.5g 10mL 6:4 30s 5min 4000r/min 5min 40 30min-40min mL 6mL 6mL 2mL 4mL 10mL : 2:3 HPLC 16 PAH Hypersil Green PAH 3µm mm A: 99:1 : B: 99:1 : 50:50-5min 50:50-25min 0:100 2mL/min 10µL UV 254nm PAH HPLC (1) (2) (3) (4) (5) (6) (7) (8) (9) (a) (10) (11) (b) (12) (k) (13) (a) (14) (a,h) (15) (ghi) (16) (1,2,3-cd) 63

65 HPLC 3 PAH Accucore C18 2.6µm mm % :45% 0.5 ml/min 2µL UV 254nm 50 3 PAH HPLC GC/MS 18 PAH TraceGOLD TG-5SiIMS 30m 0.25mm 0.25µm min 35 /min /min 320 2min 18.43min He 1 ml/min 300 1µL 1min 30mL/min MS ppb PAH 1. (j) 2. (b) 3. (k) (a) (b) (j) (k) (a) (e) (1,2,3-cd) (a,h) (g,h,i)

66 SPE-LC/MS 200mg 3mL HyperCarb ( ) 2g 15mL 10min 6000rmp 5min 3mL 1mL 1mL 3mL 1mL/min 500µL20% ppb LC/MS 2. 50ppb 121.8% LC/MS Hypercarb 5µm mm A: +5mM B: /min %A %B mL/min 25 10µL MS HESI SRM 3700V

67 SPE-LC/MS 200mg 3mL HyperSep Retain-CX ( ) 2g 0.01g 50mL 2% / 20/80 20mL 20min 4000rpm 5min 40mL mL 2% 3mL 3mL 3mL 2% 3mL 3mL 5% LC/MS Hypersil Gold 5µm mm A: 5mM B: 1. Time(min) A(%) B(%) LOQ 0.1µg/kg % 10µL 250µL/min MS ESI SRM 4500V

68 SPE-LC/MS 200mg 3mL HyperSep Retain-CX ( ) 0.5g 0.1% 10mL 1min 1mL 2% 20min 8000rpm 10min 3mL 3mL 3mL 3mL 5min 5mL 5% / 1-2mL/min LC/MS Syncronis HILIC 5µm mm A: B: 10mM / ph=3 85:15 10µL 250µL/min MS ESI MRM 3500V 350 LC/MS/MS 67

69 SPE-LC/MS 200mg 6mL HyperSep Retain PEP ( ) 4g 10mL 15min, 4000rpm 5min 10mL 4mL 4% 30:70 4mL 1min 4000rpm 3min 10mL 6mL 6mL 1-2mL/min 6mL 30% 6mL µg/kg LC/MS Hypersil GOLD TM 50mm 2.1mm 1.9µm A: B: min A% % % % 20µL 500µL/min MS ESI SRM 3000V L00 L0Q 0.05µg/kg 3. 85%-90% 68

70 SPE-HPLC 200mg 3mL HyperSep Retain-CX ( ) 5g 10mL 10min 20mL / 3:2 2min, 6000rpm 5min 20mL / 3:2 4mL / 30: mL 3mL 1-2mL/min 3mL 0.1mol/L 3mL 3mL 5% LC/MS Hypersil GOLD TM 150mm 4.6mm 5µm / / = 30:70:0.5 v/v 20µL 1mL/min UV 220nm SPE-HPLC 200mg 6mL HyperSep Retain-CX ( ) 5g 25mL 5min 6000rpm 5min 25mL 50mL 10mL 50mL 6mL 6mL 1-2mL/min 6mL 5% 5mL 6mL 5% HPLC Hypersil GOLD TM 150 mm 4.6 mm 5µm /0.05mo1/L =75/25 20µL 1mL/min UV 230nm 69

71 SPE-LC/MS 200mg 3mL HyperSep C18 ( ) 5g 30mL 10% ph 10 4g Nacl 40mL - 7:3 2min 10000rpm 5min 20mL - 7:3 5mL10% 3mL 3mL 1-2mL/min 3mL 2*2 ml HPLC Hypersil GOLD TM mm 5µm A: 0.1% B +0.1% 35:65 20µL 1mL/min UV 235nm HPLC 2. 75%-84% 70

72 SPE-LC/MS 200mg 6mL HyperSep Retain-PEP ( ) 5g 50mL 20mL r/min mL 4mL 6mL 6mL 6mL 5min 6mL 6mL 1-2mL/min LC/MS Hypersil Gold 5µm mm A: B: 0.1% 2mM 1. (min) A B (µl/min) µL 250µL/min MS SRM ESI 4500V µg/kg 2. 78%-89% 71

73 SPE-GC/MS QueChERS ( ) 20g 20mL Thermo QueChERS extraction 1min 2min 1mL 1mL Thermo QueChERS clean-up 0.5min 2min GC/MS TG-5MS 30M 0.25mm 0.25µm min7 /min min He 1.2 ml/min 20µL 10 1,290 MS GC/MS 1.naphthalene-d8 2dichlorvos 3.acenaphthene-d10 4.methacrifos 5. -BHC 6.hexachlorohenxene 7. -BHC(lindane) 8. -BHC 9.phenanthrene-d10 10.diaxinon 11.etrimphos 12.phosphamindon 13.chlorpyrifos-methyl 14.heptachlor 15.fenitrothion 16.pifimphos methyl 17.malathion 18.aldrin 19.chlorpyrifas 20.heptachlor epaxide 21.oxychlordane 22.trans-chlordane 23.endosulfan 24.cis-chlordane DDT 26.dieldrin 27.endrin 28.endosulfan 29.4,4'DDE 30.4,4'DDD 31.endosulfan sulfate 32.4,4-DDT 33.chrysene-d12 34.perylene-d12 72

74 SPE-LC/MS QueChERS ( ) 15g 30mL ACN Thermo QueChERS extraction 1min 2min 1mL 1mL Thermo QueChERS clean-up 0.5min 2min LC/MS Hypersil Gold mm 5µm A: B: 0.1% 30%A-100%A-100%A min 20µL 300µL/min MS ESI SRM 4500V 550 SPE-LC/MS QueChERS ( ) 15g 30mL ACN 30s 2min Thermo QueChERS extraction 1min 2min 5mL 5mL Thermo QueChERS clean-up 0.5min 2min HPLC HPLC Acclaim Carbamate 3µm mm A: B: min 14% 2.0min 20% 8.0min 40% min 70% 50 10µL 0.9mL/min 330nm 465nm : 1.Aldicarb sulfoxide 2.Aldicarb sulfone 3. Oxamyl 4. Methomyl 5. 3-Hydroxy carbofuran 6.Aldicarb 7.Propoxur 8.Carbofuran 9.Carbaryl 10.1-Naphthol 11.Methiocarb 12.BDMC(I.S.) 73

75 SPE-GC/MS 500mg 6mL HyperSep Florisil ( ) 10g 5g 30mL 2min 6000rpm 5min 30mL 6mL / 1:1 5mL / 1:1 10mL / 1:1 1mL GC/MS HPLC TG-5MS 30m 0.25mm 0.25µm min 25 /min 150 8min 7 /min /min min 1.0mL/min 2.0µL 290 MS MS eV SPE-HPLC 200mg 3mL HyperSep Retain-CX ( ) 5g 5mL 0.1mol/L NaOH ph mL 1min, 6000rpm 5min 20mL 10mL 0.1 mol/l HCL 3mL 3mL 1-2mL/min 3mL 2% 0.1 mol/l HCL 3mL 3mL 5% HPLC Hypersil GOLD TM 150mm 4.6mm 5µm /20mmol ph=3 =25/75 20µL 1mL/min UV 288nm 30 74

76 SPE-HPLC 500mg 3mL HyperSep Florisil ( ) 10g 5g 20mL 5min 10000rpm 5min 10mL / 10:90 3mL 1-2mL/min 6mL / 10:90 10mL / 20:80 HPLC Hypersil GOLD TM 150mm 4.6mm 5µm / =25/75 20µL 1mL/min UV 260nm 40 QueChERS ( ) 15g 80 15mL ACN / mL 1mL mL GC/MS 2 TPP 1mL mL 1mL 8µL GC/MS LC/MS mL LC TPP 0.86mL 6.7mM LC/MS 75

77 SPE-GC/MS QueChERS ( ) 15g 15mL 1% Therma QueChERSextraction 2min 3000rpm 5min, 11mL 11mL Thermo QueChERS initial clean-up 2min 3000rpm 5min 5mL mL / 9:1 1mL Thermo QueChERS final clean-up 2min 3000rpm 5min 1.GC/MS GC/MS Thermo Scientific TraceGOLD TG-5MS 30m 0.25mm 0.25µm min25 /min /min /min min He 1mL/min 2µL 290 MS Figure 2: TIC for the GC/MS analysis of grapes spiked with 1ng/µL of each pesticide % RSD n=3 21 Pesticide (peak number, name) Rt (min) Linearity (R 2 ) Average%recovery(n=3) Recovery%RSD 1. Chlorpyrifos methyl Metalaxyl Malathion Chlorpyrifos Penconazole Procymidon Dicofol Permethrin Isomer a Permethrin Isomer b Table 1: Summary of results 76

78 SPE-GC/MS QueChERS ( ) 15g 15mL 1% Thermo QueChERSextraction 5min 3000rpm 5min 11mL 11mL Thermo QueChERS initial clean-up 2min 3000rpm 5min 5mL mL / 9:1 1mL Thermo QueChERS final clean-up 5min 3000rpm 5min 1.GC/MS GC/MS Thermo Scientific TraceGOLD TG-35MS 30m 0.25mm ID 0.25µm min25 /min /min min25 /min min He 1mL/min 2µL 290 MS Figure 3 Total ion chromatogram of full scan data for 50ng/g of pesticidesin onion matrix Figure 4 Total ion chromatogram of the MS/MS analysis of 50ng/g pesticides in onion matrix 77

79 2.LOD LOQ 45 LOQ 5-78ng/g Japan 1 US-EPA 2 EU 3 EU 3 WHO 4 Component Ave.Conc. (ng/g) Std.Dev. % Recovery %RSD LOD (ng/g) LOQ (ng/g) MRL (ng/g) MRL (ng/g) MRL (ng/g) LOD 3 MRL (ng/g) Dichlorvos EPTC Mevinphos Etridazole Molinate Trifluralin Cyanophos(Thionazin) Ethoprophos Di-allate Propazine Atrazine Diazanon Gamma-BHC (Lindane) Disulfoton Heptachlor Vinclozolin Prometryn Metalaxyl Metribuzin Triadimefon Thiobencarb Dursban (Chlorpyrifos) Sevin (Carbaryl) Malathion Methiocarb Parathion Heptachlor-2,3-exo-epoxide Cyprodinil Cyanazine trans-chlordane Terbufos Sulfone cis-chlordane Endosulfan A Tetrachlorvinphos (Stirofos) p,p-dde Thiabendazole Dieldrin Chlorobenzilate Endrin Endosulfan B p,p-ddt Endosulfan Sulfate Bifenthrin Methoxychlor cis-permethrin * 100* 50* 50* trans-permethrin Average

80 % Component Avg Conc Theo Conc %Recovery %Difference %RSD Dichlorvos EPTC Mevinphos Etridazole Molinate Trifluralin Cyanophos(Thionazin) Ethoprophos Di-allate Propazine Atrazine Diazanon Gamma-BHC (Lindane) Disulfoton Heptachlor Vinclozolin Prometryn Metalaxyl Metribuzin Triadimefon Dursban (Chlorpyrifos) Thiobencard Sevin (Carbaryl) Malathion Methiocarb Parathion Heptachlor-2,3-exo-epoxide Cyprodinil Cyanazine trans-chlordane Terbufos Sulfone cis-chlordane Endosulfan A Tetrachlorvinphos (Stirofos) p,p-dde Thiabendazole Dieldrin Chlorobenzilate Endrin Endosulfan B p,p-ddt Endosulfan Sulfate Bifenthrin Methoxychlor cis-permethrin trans-permethrin Average

81 SPE-GC/MS QueChERS ( ) 10g 10mL 10mL Thermo QueChERSextraction 1min 3000rpm 5min 2mL 2mL Thermo QueChERS clean-up 1min 3000rpm 5min 1mL GC 10µL 5% 1µL 1g/mL % GC/MS Thermo ScientificTR-Pesticide 30m 0.25mm ID 0.25µm 26RF142F 60 1min15 /min 160 1min 2.2 /min min5 /min 290 5min He 1mL/min 2µL 280 MS Pesticide 0.01mg/kg mean CV 0.05mg/kg mean CV Pesticide 0.01mg/kg mean CV 0.05mg/kg mean CV Pesticide 0.01mg/kg mean CV 0.05mg/kg mean CV aldrin ethofumesate pirimiphos-ethyl benalaxyl ethoprophos pirimiphos-methyl bromopbos-ethyl etrimfos procymidone bromopropylate famoxadone propachlor bupirimate fenarimol propargite buprofezin fenazaquin propham cadusafos fenitrothion prothiofos carbofuran fenpropathrin pyrazophos chlordane-cis fenvalerate pyridaben chlordane-trans fludioxonil pyridaphenthion chlorobenzilate flusilazole pyrifenox chlorothalonil folpet quinalphos chlorpyrifos fonofos quintozene chlorpyrifos-methyl formothion simazine chlorthal-dimethyl furalaxyl tecnazene chlozolinate HCB tefluthrin DDD-pp HCH-gamma tetrachlorvinphos DDE-pp hexaconazole tetradifon DDT-op iprodione tetramethrin DDT-pp isocarbofos tolclofos-methyl deltametbrin isofenphos tolylfluanid diazinon isofenphos-methyl trifluralin dichlofluanid methacrifos vinclozolin dichlorvos methidathion dicloran methoxychlor mg/kg 0.01mg/kg dicrotophos nitrothai-isopropyl captan dieldrin oxychlordane cyfluthrin diflufenican parathion-ethyl mg/kg 0.25mg/kg endosulfan pendimethalin chlorpropham endosulfan parathion-methyl bitertanol endosulfan-sulfate permethrin cypermethrin EPN phosalone dicofol ethion phosmet diphenylamine

82 SPE-GC/MS QueChERS ( ) 10g 30mL ACN Thermo QueChERS extraction 1min 2min 5mL 5mL Thermo QueChERS clean-up 0.5min 2min GC/MS TraceGOLD TG-5MS 30m 0.25mm 0.25µm min20 /min 2805min He 1.5mL/min 1µL 280 MS GC/MS 81

83 SPE-GC/MS 500mg 6mL Carb/PSA ( ) 25.0g 50.0mL 2min 5g-7g 100mL 40mL-50mL 1min 10.00mL mL 5.0mL mL 50mL 25mL Carb/PSA mL 5.0mL 0.5mL 5.0mL GC/MS TraceGOLD TG-5MS 30m 0.25mm 0.25µm min20 /min 2805min He 1.5mL/min 1µL 280 MS GC/MS 1. 82

84 3. 5(a) (b) SIM (c) SRM 83

85 SPE-HPLC 500mg 3mL HyperSep C18 ( ) 1g +0.5mL ph4 10mL 3mL10% 10mL 2-3mL/min 5mL 15min : : =1:5:4 1:4:5 1:3:6 5mL mL 1. Rt=7.4min HPLC HPLC Hypersil Gold C18 5µm mm A : B =1:10 v/v 1mL/min UV@276nm 40 5µL % SPE-LC/MS 60mg 3mL HyperSep Retain-AX ( ) 10mL 1mL 5mL NaOH ph 11 3mL 3mL 0.1M NaOH 3mL 3mL70% 5% 3mL 1mL 3mL 89:9:2 / / 1. I-IV LC/MS 2. 73%-90% (3) (4) (5) (6) IV GC/MS Hypersil GOLD 1.9µm mm % / +0.1% 12/88 0.5mL/min 5µL 200µL/min MS ESI SRM 2500V

86 QueChERS ( ) ACN 2,000 5,000µg/mL -18 ACN MIX-1 MIX-2 10µg/mL 0.1% ACN 5µg/mL -d5-13c6 o,o - -d6 2.4-DDT -13C6 -HCH 13C6 -d10 QC -13C6 1 5µg/mL * 50mL mL 15mL 10mL ACN 200µL ISTD / mL 2mL / µL 30µL 1µg/mL QC 125µL 250 Mini-UniPrep 0.2µm PVDF LC/MS/MS 30µL QC LC GC/MS GC/MS GC/MS * 2.5g 3.5g 5.0g 5.0g A B QueChERS ( ) 5g 50mL 10mL ACN 85:15 50mL g 1g / 1 4,000rpm 10 1mL 2mL mg 50mg PSA 1 4,000rpm µm LC API 0.5mL/min %ACN% 0.1% [Na+M] m/z NIV 355 DON 319 DAS 389 HT2 447 T2 489 HPLC Betasil C18 250mm 4.6mm 5µm

87 SPE-GC/MS 1mg 6mL HyperSep Florisil SPE ( ) SPE 15mL 10mL 15mL 40mL 4min 20mL 20mL 2 1mL GC/MS TraceGOLD TG-WaxMS 30m 0.25mm 0.2µm min 5 /min /min min 1.0mL/min 10µL 250 MS eV B1 B2 G1 G2 SPE-HPLC Vicam Aflatest WB 25g 5g 100mL : 80:2 1 10mL 1 40mL 2 10mL SPE 10mL 10mL 10mL 2-4mL/min 10mL 10mL 10mL HPLC HPLC Hypercarb TM 5µm mm Dioxane/ 1:9 0.8mL/min ex=365nmem=418nm 25 5µL G2 G1 B2 B1 86

88 QueChERS ( ) 87 20mL ACN 250µL mL 2 4,500rpm 5 9.0mL / mL 1 4,500rpm 5 2.0mL µLACN 25µL 2.0µg/LQC 500µL 1 %ACN 20mM / 5 17mm 0.2µm F µL LC/MS MRM 184.0/ /126.1 S / / / / / B1b 876.6/553.4 B1a 890.7/ / / / / / / / / / / / / / / / / / / /72.1 MRM 286.9/ / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / / /131.4 A 732.6/142.2 D 746.6/ / / / / / / / /187.0 HPLC Hypersil GOLD 3µm mm

89 SPE-GC/MS 10mg 1mL SOLA HRP ( ) 1mL 1mL 200µL 200µL 200µL 200µL IS ( ) 1. 20ng/mL GC/MS 2. 20ng/mL 99.3% RSD = 2.7% n=6 Extraction Calculated Conc ng/ml Recovery% %Errorr Average Std Dev %BSD ng/mL-1000ng/mL R2= GC/MS TG-WAXMS A 30m 0.25mm 0.25µm /min min 1.2mL/min 1µL 250 1min 50mL/min MS eV Compound Abbrev SIM(start time/min) Vanillin Benzyl benzoate (Internal standard) ISTD 4.0 Quan ion (Qual Ion) 152 (151) 212 (105,91) Calculated Conc Extraction ng/ml Average 25.6 Std Dev 1.18 %BSD 4.6 Dwell time/s ng/mL RSD=4.6% 88

90 SPE-GC/MS 20mg 60mL HyperSep SLE ( ) 5 5mm 1.0g 50mL 20mL 20 min 16mL mol/L ph min 3mL 200mg/ ml min 1.5mL min 15min 20mL 4 SLE 1mL/min 35 1mL 2mL 1.GC/MS GC/MS TRACE TG-5MS 30m 0.25mm 0.25µm min 20 /min 150 8min 20 /min min 20 /min 280 1min He 1mL/min 1µL 10:1 MS µg/L 1) 2) 2,4-3) 2, 6 4) 4) 5) 3-6) 2,4,5-7) 4-8) 2,4-9) 2,4-10) 2-11) ) 4-13) 4,4'- 14) 15) 4,4'- 16) 17) 4,4'- 18) 18 ) 3,3'- -4,4' - 19) 3,3-20) 4,4'- 21) 3,3'- 22) 4,4'- - -(2- ) 23) 3,3'- 2. 2,4-58% 2,4-2,6-66% 63.8% 75% 78%-113% 89

91 AMPA SPE-LC/MS 500mg 6mL HyperSep Hypercard ( ) 6mL 6mL ph=3 5min 20mL HPLC-MS/MS LC/MS Hypercarb 5µm mm A 0.2% - B A:B=95:5 5µL 500µL/min MS ESI SRM 3500V HPLC-MS-MS 1g 6mL HyperSep Verify-CX ( ) ph 6.0 1L HyperSep Verify-AX 1 5mL 1 10mL 1 3mL/min 1 10mL mL 1mol/L HCI/ 4/1 1mL/min 50 50µL MTBSTFA* 50µL GC/MS * TS GC TraceGOLD TG-17MS 30m 0.25m 0.25µm

92 SPE-HPLC 500mg 6mL HyperSep Retain PEP ( ) 500mL 1% 6mL 6mL 1% 500mL 6mL1% 6mL HPLC HPLC Hypersil GOLD 5µm mm A: +0.1% B: + 0.1% 5%B 1.5 mins 19.5 mins 95%B 1.5 mins 0.6mL/min 10µL UV nm 60 (1) (2) 4- (3) 2,4- (4) 2- (5) 4- (6) 4- (7) 2- (8)2,4- (9) 2,4- (10) 2,4,6- (11) SPE-HPLC 500mg 6mL HyperSep Retain PEP ( ) 500mL 150g ph 6 3mL 3mL 500mL 1-2mL/min 5mL 5mL HPLC HPLC Acclaim Explosives E1 and E mm 5µm , / 5µL 1mL/min 254nm 91 1.HMX 2. RDX 3. 1,3,5-Trinitrobenzene 4.1,3-Dinitrobenzene 5. Nitrobenzene 6. Tetryl 7. 2,4,6-Trinitrotoluene 8. 4-Amino-2,6-Dinitrotoluene 9. 2-Amino-4,6-Dinitrotoluene 10.2,6-Dinitrotoluene Dinitrotoluene 12.2-Nitrotoluene 13.4-Nitrotoluene 14.3-Nitrotoluene

93 SPE-HPLC 500mg 6mL HyperSep Retain PEP ( ) 500mL + 5mL:5mL 500mL 1-2mL/min 10mL1:1 - HPLC (1) PCB-118 (2) PCB-80 (3) PCB127 (4) PCB-77 HPLC Hypercarb 7µm mm A: 30% /70% 10µL 1mL/min 254nm SPE-HPLC 500mg 6mL HyperSep Retain PEP ( ) 500mL 6mL 6mL 1% 500mL 6mL 6mL HPLC (1) (2) (3) (4) (5) (6) (7) (8) (9) (a) (10) (11) (b) (12) (k) (13) (a) (14) (a,h) (15) (ghi) (16) (1,2,3-cd) HPLC Hypersil Green PAH 3µm mm A: 99:1 : B:99:1 : 50:50-5min 50:50-25min 0:100 2mL/min 10µL UV 254nm 20 92

94 60mg 3mL HyperSep Retain PEP ( ) w/w 20% 10mL10ppm Retain PEP 2mL 2mL 1 2mL/min 4mL 1mL 5mL 0.5mL ACN/ 40:60 v/v 1mL/min 300C 214nm HPLC Hypersil GOLD 5µm mm N,N- 60mg 3mL HyperSep Retain PEP ( ) ppm N,N- Retain PEP 3mL 3mL 1 2mL/min 3mL 1mL N,N- 3mL / 95:5 v/v 1mL / 50:50 v/v HPLC Hypersil GOLD aq 5µm mm (ppm) (av) (av) Retain PEP (%)Retain PEP a b c N,N (ppm) (av) (av) Retain PEP (%)Retain PEP a b c

95 60mg 3mL HyperSep Retain PEP ( ) 10ppm 10mL 20µL Retain PEP 2mL 2mL 1 2mL/min 4mL 1mL 5mL 0.5mL ACN/0.2% 10:90 v/v 1mL/min 30 : 214nm HPLC Hypersil GOLD 5µm mm mg 6mL HyperSep Retain PEP ( ) 0.5 M H 2 SO 4 500mL ph 3 Retain PEP 3mL 5mL 5mL 5mL/min 500mL 5mL / 0.8mL PEP SPE 2 3mL 3mL 10mg 6mL HyperSep C18 ( ) 1L ph 1 C18 10mL / 50:50 10mL 5% HCI 10mL 8 10mL/min 1 HCI 10mL ph mL / 50:50 / 500µL DMF 500µL / 100µL TCTEF 1 2µL GC 2,4-D 2,4,5 Silvex 94

96 500mg 6mL HyperSep Retain PEP ( ) 500mL H 2 SO 4 ph 2 HyperSep Retain PEP 1 5mL 1 5mL 1 500mL 5mL/min 1 5mL 1 1mL 1 3mL THF 3mL HPLC HPLC Hypersil GOLD PFP 3µm mm ,4-500mg 6mL HyperSep Retain PEP ( ) H 2 SO 4 ph 2 HyperSep Retain PEP 1 5mL 1 5mL 1 500mL 5mL/min 1 5mL 1 1mL mL THF 3mL HPLC HPLC Hypersil GOLD PFP 3µm mm ,4-500mg 6mL HyperSep Retain PEP ( ) 0.5 M H 2 SO 4 500mL ph Retain PEP 3mL 5mL 5mL 5mL/min 500mL 5mL / 2,4-0.8mL PEP SPE 2 3mL THF 3mL 95

97 500mg 6mL HyperSep Retain PEP ( ) 497.5mL ph 2.5mL Retain PEP 3mL 5mL 5mL 5mL/min 500mL 5mL / 0.8mL PEP SPE 2 Retain PEP 5g 3mL 3mL 3mL 10mL / 90: mL HPLC HPLC Hypersil GOLD 3µm mm g 6mL HyperSep C18 ( ) C18 10mL / 50:50 10mL 10mL 8 10mL/min 1 20mL mL / 50:50 / 500µL DMF 500µL - 4,4'-DDE - 4,4'-DDD - 4.4'-DDT 1 60mg 3mL HyperSep Retain PEP ( ) 1 10% 2ppm 2 1% 0.2ppm Retain PEP 2mL 2mL 1 2mL/min 10mL 1mL 4mL 1% 85:15 v/v 1mL :1% 85:15 v/v 1 ml/min nm HPLC Hypersil GOLD aq 5µm mm

98 60mg 3mL HyperSep Retain PEP ( ) HyperSep Retain PEP 1 3mL 10:90 v/v 1 3mL 1 3mL 20mL/min 1L mL mL 1 2mL 10:90 v/v 1mL HPLC HPLC Hypersil GOLD PFP 3µm mm mg 3mL HyperSep Retain PEP ( ) 10ppm 10mL / 95:5 v/v 20µL Retain PEP 2mL 2mL 1 2mL/min 4mL 1mL / 5/95 v/v 4mL ACN/ 40:60 v/v 1mL/min nm HPLC GC/MS 500mg 6mL HyperSep Retain PEP ( ) Hypersil GOLD PFP 5µm mm L 6N HCI ph 2 HyperSep Retain PEP 1 3mL CH 3 CI 1 3mL 1 3mL 0.05N HCI 20mL/min 1L mL mL CH 3 CI 1 3mL CH 3 CI mL CH 3 CI 1.0mL GC/MS GC TraceGOLD TG-5MS 30m 0.25m 0.25µm

99 SPE-HPLC LC/MS 500mg 6mL HyperSep Retain PEP ( ) 0.2% 3mL 3mL 500mL 1-2mL/min 5mL 5mL HPLC Hypersil GOLD C8 5µm mm A: B: 0minsB=20% 15minsB=23% 25minsB=75% 10µL 1.5mL/min 240nm 1. HPLC LC/MS Hypersil GOLD 50mm x 2.1mm ID 3µm / 0.1% 0minsB=20% 15minsB=23% 25minsB=75% 10µL 400 µl/min MS ESI SRM 4000V 300 (1) Simazine (2) Monuron (3) Chlorotoluron (4) Atrazine (5) Diuron (6) Propazine (7) Linuron % 98

100 SPE-LC/MS 200mg 6mL HyperSep Retain PEP ( ) 500mL 6mL 6mL 5mL/min 5mL 4mL LC/MS/MS 2. 80% LC/MS Hypersil GOLD mm 3µm A: 5mM B: min A% B% µL 250µL/min MS ESI SRM 3500V

101 60mg 3mL HyperSep Retain PEP ( ) 1% 0.5 1ppm Retain PEP 2mL 2mL1% 1 2mL/min 10mL 1mL 1% 2mL 1% 1:1 1mL /1% 1:1 v/v HPLC Hypersil GOLD aq 5µm mm n= nitro phenol m , ,4, g 15mL HyperSep C18 ( ) 0.45µm 200mL 2mL 2 C mL 15 20mL 10mL 8 10mL/min 200mL 20mL mL / / 100µL 1 2µL GC

102 500mg 3mL HyperSep NH 2 ( ) 0.45µm 200mL 2mL 2 NH 2 20mL / (TCTFE) 15 20mLTCTFE mL 20mL 8 10mL/min 200mL 20mL 30 20mLTCTFE / / 100uL CTFE 1 2 GC (e) (b) (k) (a) (a,h) (g,hi) (a) (1,2,3-cd) (SPE-LC/MS) 200mg 6mL HyperSep Retain Ax ( ) 200mL 6mL 6ml 5mL/min 5mL 2% 5mL 5% LOQ 13 (LOQs) 0.5-5pg/ml LC/MS Thermo Scientific HypersiIGOLD mm, 3pm A: 0.5% +0.04% B: 0.5% +0.04% 20µL 200µL/min MS ( ESI) (SRM) 4000V

103 (SPE-LC/MS) 200mg 6mL HyperSep Retain PEP ( ) 50mL 200mL ph 4 6mL 6mL 5mL/min 5mL 4mL LC/MS/MS LC/MS Hypersil GOLD mm, 3pm / 0.1 % FA OminsB=20%, 15mins =23%, 25minsB=75% 20µL 200µL/min MS (ESI) (SRM) 4000V ng/l LC-ESI(+)-MS/MS SRM 2. LOQ:5 LOQ 11-55ng/L 3. : 72% 3.1% 19.0% 102

104 (SPE-LC/MS) 200mg 3mL HyperSep C18 ( ) 4 0.7µm 0.45µm 24 3mLMTBE, 3mL 3mL 1L 5mL/mi 3mL 10/90 MeOH/MT E3ML 3mL 1. LOQ: (LOQs) ng/L 2. : 72%~94% (%RSD) 3%~11% LC/MS Thermo Scientific HypersiIGOLD mm 3 µm A 0.1% B : T=0, A=90%, B=10% T=1, A=90%, B=10% T=15, A=1%, B=99% T=16.5, A=1%, B=99% T=17, A=90%, B=10% T=22, A=90%, B=10% 20 µl 500 µl/min MS APCI CID -10v 3 RSD (min ) LOD*(ng/L) R2** ***(%) RSD(%) a CC *LOO( )Mille Iles River R2** ng/L *** 50ng/L, n=6 103

105 (SPE-LC/MS) 200mg 3mL HyperSep C18 HyperSep Retain-CX ( , ) 1.2pm 45 µm 250mL ( 50mM) 1 ml5% Na2EDTA(w/v ) 1.0 M ph 3 3mL 3mL 1L 5mL/min HyperSepC18 3mL 2 2.5mL : (1:1), HyperSep Retain-CX, 2 2.5mL 5% : ( 1:1) 1. LOQ: ng /L 2. : 54%-105% LC/MS ThermaScientific Hypersil GOLD A (0.1% ) (min) A% B% mm, 3 µm 20pL 200pL/min MS (ESI) (SRM) 3500V mg 3mL HyperSep Retain PEP ( ) (ph 2.5) Retain PEP 5mL 5mL (ph 2.5) 1 2mL/min 10mL 3mL (ph 2.5) 5mL ACN/ (ph7.8)(9:1,v/v) 1mL :ACN- - (0.2% ) :1 ml/min :30 : (%) Retain PEP HPLC Hypersil GOLD 5pm, mm

106 LC/MS/MS 10g 75mL HyperSep C18 ( ) % H 2 SO 4 ph Z HyperSep C18 1 5mL 1 5mL 10mL/min 2 5mL 1 2mL/min <40 100µL µL LC/MS : 0.1 M (A): (B) %B ve m z-ve m z D MCPP ,4,5-T ,4,5-TP Silvex ,4-DB ,4-D, ,4-T, T, D, GC ODS-Hypersil C18 5µm, 100 2mm MDS2-Hypersil C18 3µm, 100 2mm EPA mg 6mL HyperSep Hypercarb ( ) 250mL HyperSep Hypercarb 1 20mL / 1 30mL 1 3mL 10 15mL/min 1 5mL 1 15mL / 5mL/min mL5mM / LC/MS/MS MRM OA 206/134 OA 224/152 ESA 256/80 ESA 274/80 OA 270/198 ESA 320/80 OA 264/160 OA 264/146 ESA 314/80 OA 278/206 ESA 314/80 ESA 328/80 ESA (sur) 300/80 ESA (IS) 356/80 GC Hypersil GOLD 3µm, mm

107 EPA 8330B mg 6mL HyperSep Retain PEP ( ) 2.0g 25mL 2g 0.1mL ( / ) LCS, LCSD 0.5mL 10mLACN / 1 18 (<100C) CaCI 2 (5.Og/L) 5mL 1gm 3mL 3mL 12mL 1mL 5.OmL 1 5mL ACN 1 15mL IPA 1 15mL CH 3 OH 1 5mL ACN 1 15mL ACN HyperSep Hypercarb 1 20mL ACN 1 20mL ACN 1 50mL 1 50mL 1 15mL ACN 1 30mL 10mL/min 1 4mL ACN 1 5mL ACN mL 0.7mL 100µL LC/MS : 0.50mL/min : 50:50 : GC Hypersil GOLD 5µm, mm Hypersil GOLD C8 3µm, 150 4mm Hypersil GOLD CN 5µm, mm Betasil Phenyl Hexyl 5µm, 250 3mm

108 EPA g 75mL HyperSep C18 ( ) 1L MgCI 2 ( 10mg/L ) HyperSep C ml 1:1 EtAc/CH 3 Cl mL 1 10mL 1 5mL 50uL 1 2mL/min 1 10mL EtAc 1 10mL CH 3 Cl mL 1mL 1 2µL GC GC TraceGOLD TG-OCP I 30m 0.25m 0.25µm mL EtAc/CH 3 Cl 2 SPE HyperSep Aminopropyl, HyperSep Retain-AX, HyperSep SAX HyperSep Verify-AX HyperSep 1mL 1 3mL 1 3mL 1 3mL/mi 10 50mL 1 10mL ( >10 5 ) 100mM 1 3mL (100mM) 1 3mL/min 1 3mL 1 3mL/min 107

109 4' 4-100mg 1mL HyperSep C18 ( ) Verify-CX 3mL 3mL 1 ml/mi 1mL 4' 4 1M 0.25mL 10µL HPLC 100µg/mL :3.162 : / (50:50) :1.2mL/min :10µL : (254nm) GC Hypersil GOLD 5pm, mm mg 3mL HyperSep Retain PEP ( ) 100mL 10mg 100ppm 25mL (100ppm) 50mL ( 50ppm) 25mL (100ppm) 50mL 1%H 3 PO 4 ( 50% 1 %H 3 PO 4 ) Retain PEP 2mL 2mL 2mL 2mL / (5/95, v/v) (5 10 >10 / ) 2mL <50 1mL 60mg 3mL HyperSep Retain-CX ( ) 100mL 10mg 100ppm 25mL (100ppm) 50mL ( 50ppm) 25mL (100ppm) 50mL 1%H 3 PO 4 ( 50% 1 %H 3 PO 4 ) Retain PEP 2mL 2mL 2mL 2mL / (5/95, v/v) (5 10 >10 / ) 2mL <50 1mL 108

110 Hyclone 60mg 6mL Retain PEP CX ( ) 100mL 10 - (100ppm) 25mL - 50mL 25mL - 50mL 1 2mL CH 3 OH 1 2mL H 2 O 2mL 2mL 1 2mL 5% CH 3 OH 1 2mL CH 3 OH SPE (1%) 1 2 HyperSep Retain PEP+50% HyperSep Retain-CX+50% HPLC 200mg 6mL HyperSep Verify-CX ( ) 1 ml 100mM (ph 6.0) * 1mL / ph mM ph HyperSep Verify-CX 1 3mL 1 3mL 1 1 ml 100mM (ph= 6.0) <3 1mL/min 1 2mL 1 2mL 20% 100mM (ph= 6.0) ( >10 10 ) 1 2mL 1 5mL 2% 1 2mL/min <40 HPLC * -D5 Alpha- - -D5 -D4 -D5 -D5 GC BETASIL Phenyl/Hexyl 5µm, mm

111 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) - 2mL 1mL mM (ph=5.0) F /ml Patella Vulgata / ,000rpm 10 HyperSep Verify-CX 1 3mL 1 3mL 1 1 ml 100mM (ph= 6.0) <3 1mL/min 1 2mL 1 2mL20% 100mM (ph 6.0) ( >10 5 ) 1 2mL 1 5mL 4% 1 2mL/min <40 50NL 50}L STFA** ( 1%TMCS}** / BSTFA GC/MS 1-2µL *** -D5-TBDMS TBDMS D5-TBDMS TBDMS Serax Serax D5-TBDMS TBDMS Restoril TBDMS Ativan Klonopin TBM Valium TBDMS * TBDMS Versed TBDMS TBDMS Xanax D5-TBDMS TBDMS Halcion TMS * GC/MS -D5 ** TS *** GC TraceGOLD TG-5MS 30m 0.25m 0.25µm

112 : GC-GC/MS 200mg 6mL HyperSep Verify-CX ( ) 1 ml 100mM (ph= 6) * 1mL / 1g}1:4) / 3mL 100mM (ph=6) ph mM ph / 1mL 5000 F /m L b- (ph=5.0) * 1mL / ,000rpm 10 3mL 100mM (ph 6.0) HyperSep Verify-CX 1 3mL 1 3mL100m M (ph 6) <3 1 2mL/min 1 3mL 5% p/v) 100mM (ph6) ( >10 5 ) 1 3mL ( >10 5 ) 1 3mL (98:2 v/v) 1 2mL/min <40 500L 50µL STFA( 1 % TMCS**) GC/MS D* Alpha Alpha- - -D5* D5* D4* D5* D5* D5* * -D Alpha- - -D5 -D4 -D5 -D5 -D5 ** TS GC TraceGOLD TG-5MS 30m 0.25m 0.25µm

113 : LC/MS/MS 200mg 6mL HyperSep Verify-CX ( ) 1mL100m M (ph=6) * 1mL / 1g(1:4) / 3mL 100mM (ph=6) ph mM ph / 1mL 5000 F /ml b- (ph=5.0) 1mL / 650C 3 2,000rpm 10 3mL 100mM (ph 6.0) HyperSep Verify-CX 1 3mL 1 3mL 100mM (ph=6) <3 1 2mL/min 1 3mL 5% w/v) 100mM (ph6) ( >10 5 ) 1 3mL ( >10 5 ) 1 3mL ; (98:2 v/v) 1 ZmL/min <40 50µL 0.02% ( ) LC/MS/MS 30:70( :0.02% ) :0.35mL/min : : 5µL MS MRM 309.1/ D* 314.1/286.2 Alpha /242.9 Alpha- - -D5* 330.1/ / /192.5 * 292.2/ / D4* 325.1/ / D5* 275.6/ / D5* 290.2/ / D5* 306.1/260.1 * -D Alpha- - -D5 -D4 -D5 -D5 -D5 GC Hypersil GOLD PFP 3pm, mm

114 Beta GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1 ml 100mM (ph 4.5) 1mL 2mL 100mM (ph 4.5) / HyperSep Verify-CX 1 3mL 1 3mL 1 3mL 100mM (ph 4.7) <3 1 2mL/min 2 1mL / (1:1) ( >10 5 ) Beta 1 1mL / (78:20:2) 1 2mL/min ( ) IPA/NH 4 0H (ph 11-12) <40 : ( ) 5mg/mL -20 ( ) 100µL ( ) / µL ( ) GC/MS GC TraceGOLD TG-5MS 30m 0.25m 0.25µm GC/MS 200mg 10mL HyperSep Verify-CX ( ) :B- 2mL 1mL - 100mM (ph5.0) F L Patella Vulgata / 65 3 HyperSep Verify-CX 1 3mL 1 3mL 1 1 ml 100mM (ph 6.0) <3 1 2mL/min 1 2mL 1 2mL 20% 100mM (ph=6.0) ( >10 5 ) 1 2mL /7-1 3mL 2% NHqOH 1 2mL/min <40 50µL 50uLMTBSTFA( 1% TBDMCS) / MTBSTFA 1 2µL GC/MS *** -TBDMS TBDMS D4-TBDMS D4-TBDMS * GC/MS -D4 7- -D4 ** TS *** GC TraceGOLD TG-5MS 30m 0.25m 0.25µm

115 Beta GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL (ph 4.5) 1mL 2mL (ph=4.5) / HyperSep Verify-CX 1 3mL 1 3mL 1 3mL100m M (ph 4.5) <3 1 2mL/ 2 1mL / (1 :1) ( >10 5 ) Beta 1 1mL / / (78:20:2) <40 : ( ) 5mg/mL 200C( ) 100µL ( ) / GC/MS GC TraceGOLD TG-5MS 30m 0.25m 0.25µm IPA/NH 4 0H (ph 11-12) / 200mg 10mL HyperSep Verify-CX ( ) 1 ml 100mM * 1mL / 2mL 100mM / HyperSep Verify-CX 1 3mL 1 3mL 1 1 ml 100mM <3 1 2mL/min 1 3mL 1 3mL10 mm ( >10 5 ) / / 1 3mL : (90:10) 1 2mL/min < µL 0.1% ( ) 20µL LC * 8- GC Hypercarb 3µm, mm

116 DHEA, GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 5mL ph ,000rpm 5 HyperSep Verify-CX 1 3mL 1 3mL 1 1 ml 100mM (ph 6.0) 1 3mL ( >10mm ) 1 3mL 1 2mL/min 2mL 200mM (P 7.0) 250 p / 500C 1 1; 1 NaHC0 3 /Na 2 C0 3 ph mL n rpm pLMSTFA/NH41/ (1,000:2:5, V/W/W) rpm 1 GC 1 2µL GC/MS ** DHEA a * * GC/MS 20ng/mL 16a ** GC TraceGOLD TG-5MS 30m 0.25m 0.25µm GC GC/MS 100mg 1 ml HyperSep Diol ( ) 1mLp 6 2mL / 5m1 ph rpm 15 HyperSep Diol 1 3mL 1 3mL 1 3mL 1 3mL 0.1mM (ph= 6.0) 1 3mL 1 3mL5% 0.1 mm (ph=6.0) > mL 2 3mL µL 100µL 1 % t-bdmcs MTBSTFA GC/MS-NCI 1µL MTBSTFA * -D5 -D4 GC TraceGOLD TG-5MS 30m 0.25m 0.25µm

117 60mg 3mL HyperSep Retain PEP ( ) 100mL 10mL (20mg/L) 30mL 0.5% 100mL 2ppm Retain PEP 2mL 2mL 2mL 2mL 5% 0.5% 2mL1% ACN:20mmol (ph 4) (40:60, v/v) 1mL :ACN:20mmol (ph 4) (40:60, v/v) :1.0mL/min :10µL :30 :290nm GC Hypersil GOLD 5pm, mm mg 3mL HyperSep Retain PEP ( ) HyperSep Retain -PEP 1 3mL 1 3mL S E 1 3mL 1 3mL 5% 1 3mL (%) GC/MS 500mg 6mL HyperSep SAX ( ) 1mL 100pL D3-MMA 1mL / rpm 5 1 3mL 1 3mL SPE 1 10mL 3 1 5mL 3 1 2mL MTBE* 3 1 5mL3% MTBE 1 2 ml/min <35 25µL MSTFA+1%TMCS 256L µL GC/MS GC TraceGOLD TG-5MS 30m 0.25m 0.25µm

118 / GC GC/MS 100mg 1mL HyperSep C18 ( ) 500µL 20% * / 500µL / / HyperSep C18 1 3mL 1 3mL 1 1 ml 100mM HCI <3 1 2mL/ 1 3mL 1 3mL 1 3mL ( >10 5 ) 1 1mL2% <40 50µL 50µL BSTFA( 1 % TMCS ) **500L µL GC/MS -TMS D10-TMS* * (FID): -D10(GC/MS) ** TS GC TraceGOLD TG-5MS 30m 0.25m 0.25µm / LC/MS 200mg 6mL HyperSep Verify-CX ( ) mL 1mL ( ) * / 3mL 100mM HCI / HyperSep Verify-CX 1 3mL 1 3mL 1 1 ml 100mM HCI <3 1 2mL/min 1 3mL 1 3mL 1 3mL ( >10 10 ) 1 3mL 2% 1 2mL/min < GC/MS * -D10 - GC Hypersil GOLD 3pm, mm

119 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL 100mM (ph =6.0) * 2mL ph mM ph / HyperSep Verify-CX 1 3mL 1 3mL 1 1 ml 100mM (ph 6.0) <3 1mL/min 1 3mL 1 2mL200mM HCI ( >10 5 ) 1 2mL 1 3mL ( >10 5 ) 1 3mL /IPA/NH 4 OH (78:20:2) 1 ml/min IPA/NH 4 0H (ph 11-12) <40 100µL 1 2µL GC/MS *** D4* D3* * -D4 -D3 ** GC TraceGOLD TG-5MS 30m 0.25m 0.25µm mg 3mL HyperSep Cyano ( ) 1mL 1mL 8mL / HyperSep Cyano 1 3mL 1 3mL <3 1 2mL/min 1 3mL1% ( ) ( >10 5 ) 2 3mL1% 1 2mL/min <40 100µL 0.1% ( ) 50µL LC-UV/260nm) * GC Hypersil GOLD 3pm, mm

120 200mg 3mL HyperSep Retain-AX ( ) 3% 500µL HyperSep Retain-AX 1 1mL 1 1mL 0.5mL 1 1 ml 1% CH403S 1mL 1 1mL ACN(1% ) LC/MS/MS ACN:3mmol/L 85:15) MRM 281.2/ /315.2 C /269.2( C17 ) GC Hypersil GOLD 5µm, mm GC GC/MS 200mg 6mL HyperSep Verify-CX ( ) 2mL 400µL 200µL 10% / ,000rpm µl 50% / 50% ph mL 1mL mM (ph 5.0) F /ml Patella Vulgata ,000rpm N NaOH ph mL 1 3mL 1 1 ml 100mM (ph 6.0) <3 1 2mL/min 1 3mL 1 3mL 100mM (ph 4.5) 1 3mL ( >10 5 ) 1 3mL / / (84:12:4) <40 200µL1:1 / <40 50µL / (70:30) 1 2µL GC/MS *** D3* D3* * GC/MS -D3 -D3 ** GC TraceGOLD TG-5MS 30m 0.25m 0.25µm

121 - GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL 400µL 200µL 10% / ,000rpm pL50% / 50% ph 5-6 2mL / 60 ( ) 200µL10% ph 5-6 2,000rpm mL 1 3mL 1 3mL 100mM (ph 6.0) <3 1 2mL/min 1 3mL 1 3mL 100mM (ph 4.5) 1 3mL ( >10 5 ) 1 3mL /IPA/NH 4 OH (76:20:4) 1 2mL/min IPA/NH 4 0H (ph 11-12) <40 100µL 100µL BSTFA ( 1%TMCS}** / MTBSTFA 1 2µL GC/MS *** -D D4 TMS* TMS* TMS O- TMS N- TMS TMS D3 TMS* D6 TMS* TMS TMS TMS D3 TMS* D6 TMS* TMS TMS D3 TMS D6 TMS TMS D3 TMS TMS D3 TMS D6 TMS TMS D3 TMS TMS * GC/MS D4- D4- D3- D3- D6- D6- D6- D6- LOD/ LOQ ** TS

122 GC/MS 200mg 6mL HyperSep Retain-CX ( ) ( ) 1mL 1.0mL - ( - 100mM (ph 5.0) F /ml Patella Vulgata ) 600C T N NaOH ph ( ) 1mL 1mL 100m M (ph 6.0) 1 2mL/min 1 1mL 1 1 ml 100mM (ph 4.5) 1 1mL ( >10 3 ) 2 0.5mL / IPA/NH 4 OH (78/20/2) 1 2mL/min <40 50µL 50µL BSTFA( 1 % TMCS**), / MTBSTFA 1 2µL GC/MS : ( ) -TMS , 343 -D6-TMS* , 349 -TMS , 414 -D6-TMS* , TMS , D6-TMS , 343 * -D6-TMS -D6-TMS ** TS GC TraceGOLD TG-5MS 30m 0.25m 0.25µm / LC/MS/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL100m M (ph=6) * 1mL / 2mL 100mM (ph=6) / HyperSep Verify-CX 1 3mL 1 3mL 1 3mL 100mM (ph=6) <3 1 2mL/min 1 3mL 1 3mL 100mM 1 3mL ( >10 5 ) 1 3mL : : (78:20:2) 1 2mL/min <40 100µL : 5 LC/ MS MRM 330.0/90.1 -D6* 336.0/76.1 * -D6 GC Hypersil GOLD 3µm, mm

123 60mg 3mL HyperSep Retain PEP ( ) 100mL 10mL (100mg/L) 30mL 0.5% 100mL 10ppm Retain PEP 2mL 2mL 2mL 2mL 5% 0.5% 2mL 1% ACN:20mmol (ph 4) (30:70, v/v) 1mL :ACN:20mmol (ph 4) (40:60,v/v) :1.0mL/min :1µL :30 : (290nm) GC Hypersil GOLD 5µm, mm / HPLC 200mg 10mL HyperSep Verify-CX ( ) 4mL 2mL 100m M (PH=6.0) 1mL / / 2,000rpm 10 ph mM ph HyperSep Verify-CX 1 3mL 1 3mL 1 1mL100m M (ph=6.0} <3 1 ml/min 1 3mL 1 1mL 100mM 1 3mL ( >10 5 ) 1 3mL /IPA/NH 4 OH (78:20:2) 1 ml/min IPA/NH 4 0H (ph 11-12) <40 200µL : (1:3) U / µL LC 235nm : 30% 0.25 M (ph 2.7) :2mL/min * : 122

124 200mg 6mL HyperSep Retain PEP ( ) 50mL 50mL 0.1 M ZnS0 4 / 500mL * / 500mL / HyperSep Retain PEP 1 2mL 1 2mL <3 1 2mL/min 1 2mL ( >10 20 ) 750mL 1 2mL/min LC * -D, GC Hypersil GOLD 3µm, mm / HPLC 200mg 10mL HyperSep Verify-CX ( ) 2mL 100mM (ph=6.0) * 1mL / / 2,000rpm 10 ph mM ph 1 3mL 1 3mL 1 1 ml 100mM (ph= 6.0) <3 1 ml/min 1 3mL 1 1 ml 100mM 1 3mL ( >10 5 ) 1 3mL /IPA/NH 4 OH (78:20:2) 1mL/min IPA/NH 4 0H (ph 11-12) <40 200µL : (1:3) / µL HPLC * 123

125 (SPE-LC/MS ) 10mg 2mL SOLA HRP 96- ( ) : 1mL 1 ml 1mL 200µL / (80; 20) 200µL µL : 1. 25ng/mL LC/MS LC/MS : Accucore RP-MS 2.6um mm : : A: ++0.1% B: +0.1% : /min % A %B : 0.6 ml/min : 25 : 2.5µL MS : ( HESI) (SRM) 3000V (1) (2) (3) (4) IS(5) % RSD<12 %(n=6) 3.1ng/mL-1000ng/mL R2>

126 - (SPE-LC/MS ) 10mg 1 ml SOLA SCX ( ) : 500µL 500µL 200µL 250µL +0.1% 250gL + 0.1% 250µL DCM/IPA+S% (80; 20) 40 : ( 80; 20 ) 200µL : 1.0.5mg/mL 5 - HPLC LC/MS : Accucore C18 2.6µm mm : : A: +0.1% : +0.1% : /Min %A %B : 0.7 ml/min : 45 : 1µL UV: 220nm (1) (2) (3) (4) (5) mg/mL5-80% RSD<5 %(n=6) 125

127 (SPE- HPLC) 10mg 1 ml SOLA SCX ( ) 1 ml 1 ml 350µL 350µL +2% 1 350µL 2 350µL +5% µL 350µL HPLC : Accucore R MS 2.6um50 3 mm : : A: 20mM : : /min % A %B : 0.8 ml/min : 25 : 10µL UV: 254nm : 1. 10µg/mL HPLC IS IS 126

128 2. 10µg/mL HPLC IS IS % RSD<11.8%(n=3) 127

129 (SPE- HPLC) 10mg 1 ml SOLA SCX ( ) 1 ml 1 ml 250µL ++1S 250µL +2% 250gL +2% 250µL +5% µL : 1. 10µg/mL IS LC 1) 2) % RSD = 4.03%(n=6) HPLC : Syncronis C18 50m mm : : A: 20mM B: : /min % A %B : 1 ml/min : 30 : 10µL UV: 277nm 6 - (SLE-LC/MS) 200mg 3mL HyperSep SLE ( ) : 400µL +IS( -D3) : 5min : 1.4mL 0.5-1% 1 ml/min µL : % RSD <12% LC/MS : Hypersil Gold 5pm mm : : A: +0.05% B: +0.05% : /min %A %B : 1mL/min : 40 : 10µL MS : 128

130 25-OH Vit D2 25-OH Vit D3 (SPE- LC/MS) 10mg 2mL SOLA HRP 96 ( ) µL 2% 200µL 200µL / (60:40) 200µL 2 40 : ( 30; 70) 100µL LC/MS : Syncronis C18 1.7µm mm : : A: +0.1% B: +0.1% : 2min %B : 0.8mL/min : 40 : 50µL MS : : OH Vit D2 25-OH Vit D3 >94% RSD<4.0% LOD 5ng/mL Vit D2 25- Vit D ng/mL r 2 =

131 (SPE- LC/MS) 10mg 2mL SOLA SCX 96 ( ) 500µL 500µL 2% 2000L +IS( ) 200µL 2% 200µL 2% / (70.30) 200µL 5% µL : 1. 30ng/mL LC/MS LC/MS : Accucore RP-MS 2.6µm mm : : A: +0.1% B. +0.1% : /min % A %B : 0.35 ml/min : 40 : 5µL MS : (ESI) (SRM) 3000V ng/mL r 2 = %,RSD=6.81% 130

132 (SPE- LC/MS) 10mg 1 ml SOLA HRP ( ) 0.5mL 0.5 ml 100µL 200µL / (80; 20) 250µL µL LC/MS : Accucore PFP 2.6µm mm : : Accucore PFP 2.6µm mm : : A: B: : /min % A %B : 1mL/min : 40 : 10µL MS : ( ESI) (SRM) 2500V 300 : ng/mL LC/MS %, RSD=2.3%/n=3) 3. 10µg/mL-1000ng/mL R=

133 (SPE- LC/MS) 10mg 2mL SOLA HRP 96 ( ) 1 ml 1 ml 100µL 500µL +0.1% 500µL 200µL 90% 2 40 : ( 80; 20) 200µL LC/MS : Accucore RP-MS 2.6µm mm : : Accucore RP-MS 2.6µm mm : : A: +0.1% B: +0.1% : %B 0min 5 0.1min 5 1.0min min min 5 3.6min 5 : 0.75mL/min : 60 t: 15µL MS : (ESI) (SRM) 3000V

134 : 1. 1ng/mL LC/MS %, RSD=4.88(n=3) 3. MF % RSD<3.72(n=6) 4. 1ng/mL-1000ng/mL R 2 = ng/mL 133

135 (SPE- LC/MS) 10mg 1 ml SOLA SCX ( ) 1 ml 1 ml 100µL +1S1 ) 200µL +0.1% 250µL +3% 40 : ( 80; 20) 100µL : ng/mL LC/MS LC/MS : Accucore aq 2.6µm mm : : A: +0.1% B: +0.1% : 2min 20%-70%B : 0.4mL/min : 40 : 2.58L MS : ( ESI) ( ) (SRM) 3000V ng/mL LC/MS % 65.8% RSD<6.11n=3) ng/mL - 500ng/mL R 2 =

136 (SPE-LC/MS) 10mg 1 ml SOLA HRP ( ) 0.5mL 0.5mL 200µL 200µL / (90; 10 ) 200µL 2 40 : ( 50; 50) 100µL : 1. 1ng/mL LC/MS LC/MS : Accucore RP-MS 2.6µm mm : : A: +0.1% B: +0.1% : /min % A %B : 0.6mL/min : 40 t: 20µL MS : ( HESI) (SRM) 3000V % RSD<5 %(n=3) 3. 1ng/mL-1000ng/mL R 2 =

137 (SLE-LC/MS) 500mg 3mL HyperSep SLE ( ) : 300gL +300µL 0.1% +20µL IS(d6- ) : 5min : C 200µL : ( 80:20) : ng/mL LC/MS LC/MS : Accucore C18 2.6µm mm : : A: +0.1% B: +0.1% : /min % A % B : 0.4mL/min : 40 : 100L MS : (HESI-II) (SRM) 3500V %, RSD=1.86%(n=6) ng/mL-5ng/mL R 2 =

138 (SPE- HPLC) 10mg 1 ml SOLA HRP ( ) 0.5mL 0.5mL 200µL +IS( ) 200µL / (90:10 ) 200µL +0.1% : 1. 20ng/mL ( ) HPLC HPLC : Hypersil GOLD 3µm mm : : A: 25mM B: : /min % A %B : 1.5mL/min : 20 : 25µL : :480nm :560nm IS( ) 3. 1 ng/ml-100ng/ml R 2 = ng/mL ( ) 101.2%, RSD=6.9%(n=6) 137

139 (SPE-LC/MS ) 10mg 2mL SO A R ( ) 0.5mL 0.5 ml 200µL 200µL / (70; 30) 250µL +1% 2 40 : ( 50; 50) +0.5% 20µm 200µL : ng/mL LC/MS LC/MS : Accucore RP-MS 2.6µm mm : : A: +0.1% B: +0.1% : /min % A % B : 0.6mL/min : 30 : 2.5µL MS : ( HESI) (SRM) 4000V % RSD<9 %(n=6) ng/mL-10ng/mL,R 2 =

140 6-200mg 10mL HyperSep Verify-CX ( ) 1 x 2mL 100mM (ph= 6.0) * / 4mL 2000rpm 10 ph mM ph 6 HyperSep Verify-CX 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph=6.0) <3 1mL/min 1 x 3mL 1 x 2mL 100mM (ph 4.5) 1 x 3mL ( >10 10 ) 1 x 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 50µL / 50µL BSTFA 1% TMCS ** / BSTFA 1 2µL GC/MS *** D6-6-AM-TMS* AM-TMS * GC/MS ** TS GC TraceGOLD TG-5SilMS 30m 0.25m 0.25µm mg 10mL HyperSep Verify-CX ( ) 139 1mL 1.0mL 100mM (ph 6.0) * 1mL / ph mM ph 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph= 6.0) <3 1mL/min 1 x 2mL 1 x 2mL 20% ACN 100mM (ph 6.0) ( >10 10 ) 1 x 2mL 1 x 5mL 2% 1 2mL/min <40 HPLC * -D5 Alpha- - -D5 -D4 -D5 -D5 HPLC BETASIL Phenyl/Hexyl 5µm, mm

141 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL * 1mL 100mM (ph 6.0) 1mL 0.35 M / 20 ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 1mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) 30µL DMF*** <40 30µL 50µL PFPA (PFAA)**** 50µL PFPOH <40 1 2µL (TMS ) *** D 5 - * D 5 - * * ** *** TS mL **** TS x 1mL GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 500mg 6mL HyperSep Diol ( ) 1mL ph 6 * 2mL / 5mL ph rpm 15 1 x 3mL 1 x 3mL 1 x 3mL 1 x 3mL 0.1mM (ph= 6.0) 1 x 3mL 1 x 3mL 5% ACN 0.1mM (ph 6.0) > x 3mL 2 x 3mL 55 * 100µL ACN 100µL 1% t-bdmcs MTBSTFA µL GC/MS-NCI MTBSTFA GC/MS * -D5 -D4 GC TraceGOLD TG-5SilMS 30m 0.25m 0.25µm

142 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 5mL 2mL - - :100mM (ph=5.0) 5,000 F /ml Patella Vulgata / ,000rpm µL 1.0N NaOH ph mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 10% (v/v) ( >10 5 ) 1mL / (50:50) a 3mL /IPA/NH OH (78:20:2) 1 4 2mL/min IPA/NH OH 4 (ph 11-12) b 3mL /IPA (80:20) c 3mL d 3mL <40 50µL 50µL MSTFA( 3% ) / MSTFA 1 2µL (GC/MS) (TMS ) * -TMS TMS ,143-2TMS TMS # # TMS Alpha TMS * GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 141

143 / 60mg 3mL HyperSep Retain-CX ( ) 1mL 100mM (ph= 6) * 1mL 2mL 100mM (ph= 6) / ph mM ph / 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph= 6) <3 1 2mL/min 1 x 3mL 1 x 3mL 1% 1 x 3mL ( >10 5 ) 1 x 3mL ACN (78:20:2 v/v) 1 2mL/min <40 100µL 5µL GC/MS MRM -TMS TMS TMS TMS 418 #1 640 # TMS 405 Alpha TMS * HPLC Hypersil GOLD 3µm, mm SPE-HPLC 30mg 1mL HyperSep Retain- CX ( ) 1mL 1mL 1mL, 20µl 5N HCl 2mL 0.1N HCl, 2mL 2mL 2% 40:60 HPLC Hypercarb 5µm,100x4.6mm A: (ph9.0): (4:6) 1mL/min 10µL 220nm 30 (1) (2) -3- (3) -6- (4) (5) 142

144 / LC/MS/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6.0) * 1mL 2mL 100mM (ph 6.0) / ph mM ph / 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 3mL 1% 3mL ( >10 5 ) 3mL (78:20:2, v/v/v) 1 2mL/min <40 100µL 5µL LC 0.1% (33:67, v/v) 0.35mL/min HPLC Hypersil GOLD 3µm, mm /91.1 -D3* 281.2/ / D3* 259.2/ /107.1 EDDP 278.2/234.2 EDDP-D3* 281.4/ / D9* 319.2/ / /44.1 -D5* 331.1/ /58.1 -D11* 351.3/ / /58.1 -D3* 268.2/ / /235 * 143

145 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL * 1mL 100mM (ph 5.0) / ph mM ph 3mL 3mL 1mL 100mM (ph 5.0) <3 1 2mL/min 3mL 1mL 100mM ( >10 5 ) 2mL 3mL / (50:50) 1 2mL/min <40 100µL 25 50µL 0.2M TMPAH*** 1 2µL (GC/MS) *** * *** D C D * ** *** TS MethElut 12x1mL GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 144

146 200mg 10mL HyperSep Verify-CX ( ) 2mL 1mL mM (ph=5.0) 5,000 F /ml Patella Vulgata / ,000rpm 10 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph= 6.0) <3 1mL/min 1 x 2mL 1 x 2mL 20% ACN 100mM (ph= 6.0) ( >10 5 ) 1 x 2mL 1 x 5mL 4% 1 2mL/min <40 50µL 50µL BSTFA** ( 1% TMCS)** / BSTFA 1 2µL GC/MS *** Xananx TMS Librium Klonopin Valium TMS TMS TMS Ativan TMS TMS Serax * TMS Restoril Halcion TMS * GC/MS -D5 ** TS GC TraceGOLD TG-5SilMS 30m 0.25m 0.25µm

147 200mg 10mL HyperSep Verify-CX ( ) 2mL * 1mL - - :100mM (ph=5.0) 5,000 F /ml Patella Vulgata / ,000rpm µL GC/MS *** -D5-TBDMS TBDMS D5-TBDMS TBDMS Serax D5-TBDMS TBDMS Restoril Ativan Klonopin TBMS Valium TBDMS * TBDMS Versed TBDMS TBDMS Xanax D5-TBDMS TBDMS Halcion TBDMS * ** GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 146

148 200mg 3mL HyperSep Diol ) 1mL 100mM (ph= 6) * 1mL / 1g (1:4) / 3mL 100mM (ph= 6) ph mM ph / 1mL 5000 F /ml - (ph 5.0) * 1mL / ,000rpm 10 3mL 100mM (ph 6.0) HyperSep Diol 1 x 3mL 1 x 3mL 100mM (ph 6) <3 1 2mL/min 1 x 3mL 5% (v/v) ACN 100mM (ph6) ( >10 5 ) 1 x 3mL ( >10 5 ) 1 x 3mL (98:2 v/v) 1 2mL/min <40 50µL ACN 50µL BSTFA( 1% TMCS**) µL GC/MS D* Alpha Alpha- - -D5* D5* D4* D5* D5* D5* * -D Alpha- - -D5 -D5 -D4 -D5 -D5 -D5 GC TraceGOLD TG-5SilMS 30m 0.25m 0.25µm

149 Beta 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 4.5) 1mL 2mL 100mM (ph 4.5) / 1 x 3mL 1 x 3mL 1 x 3mL 100mM (ph 4.7) <3 1 2mL/min 2 x 1mL / (1:1) ( >10 5 ) 1 x 1mL / (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 5mg/mL ( ) -20 ( ) 100µL ( ) / µL Beta GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 4.5) 1mL 2mL 100mM (ph 4.5) / 1 x 3mL 1 x 3mL 1 x 3mL 100mM (ph 4.7) <3 1 2mL/min 2 x 1mL / (1:1) ( >10 5 ) 1mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 5mg/mL ( ) -20 ( ) 100µL ( ) / µL 148

150 - SPE-HPLC 30mg 1mL HyperSep Retain- PEP ( ) 1mL 1mL 1mL, 0.5% 1mL 0.5% 5% 1mL 1% - HPLC (1) (2) (3) (4) (5) (6) (7) HPLC Hypersil GOLD 5µm, 150x4.6mm A: + 0.1% ; B: +0.1% ; 15 min 5-55%B 1mL/min 10µL 220nm 25 GHB 200mg 3mL HyperSep Retain-AX ( ) 1mL * 0.5mL 100mM (ph 6.0) / rpm 10 HyperSep Retain-AX 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph 6.0) < x 2mL /NH OH (99:1) 4 1 / µL 1mL rpm µL 25µL BSTFA ( 1% TMCS**) / µL GC/MS * GHB-D6-di-TMS* GHB-di-TMS * GC/MS D6-GHB ** TS GC TraceGOLD TG-5SilMS 30m 0.25m 0.25µm

151 SPE- HPLC 10mg 1mL SOLA HRP ml, 1 ml 200 µl +IS 1mL / % 0.2mL + 0.1% 40 / µL Accucore C µm 50x2.1 mm A: + 0.1% B: + 0.1% /min % A %B ml/min 40 10µL 245nm 1. 10µg/ml IS HPLC RSD 5.6%(n=5) Extracted sample # Doxepin Peak Area Protryptyline (IS) Peak Area Response ratio Calculated Amount/ ug.ml-1 Recovery Mean RSD 56% Calibration Standard (5.2 ug/ml)

152 SPE- HPLC 200mg 3mL HyperSep SLE µL 5min 1.4mL 0.5-1% 1mL/min µL Hypersil Gold 5µm,150x4.6mm,Uniguard Hypersil gold & holder , , A: 0.05% ; B: 0.05 : %B,20min 1 ml/min 10µL 40 MS 500mg 6mL HyperSep Aminopropyl ( ) 151 1mL 100mM (ph= 5) * / 1mL / 2mL 100mM (ph= 5) / ph mM ph 1mL 100mM * 1 5mL / 2mL 100mM (ph= 5) helix pomatia 5,000 /ml x 3mL 1 x 3mL 1 x 1mL 100mM (ph= 5.0) <3 1 2mL/min 1 x 2mL 1 x 3mL 100mM (ph= 5.0) 1 x 3mL 5 10 >10 / ) 1 2mL/min BSTFA <40 50µL 50µL BSTFA( 1% TMCS)** BSTFA 1 2µL GC/MS -D4-TMS* TMS TMS D3-TMS* * -D4-TMS -D3-TMS ** TS GC TraceGOLD TG-5SilMS 30m 0.25m 0.25µm

153 GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 5.0) * / 1mL 2mL 100mM (ph 5.0) / ph mM ph 1mL 100mM * 1 5mL / 2mL 100mM (ph 5.0) helix pomatia 5,000 /ml 60 3 Verify-CX 3mL 3mL 3mL 100mM (ph 5.0) <3 1 2mL/min 2mL 3mL 100mM (ph 5.0) 3mL (5 10 >10 / ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) BSTFA <40 50µL 50µL BSTFA ( 1% TMCS)*** BSTFA 1 2µL GC/MS -D4-TMS* TMS TMS D3-TMS* * ** ** TS x 1mL GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 152

154 / HPLC 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 5.0) * 1mL / 2mL 100mM (ph 6.0) / ( / ) 1mL 5000 F /ml b- (ph 5.0) * 1mL / mL 100mM (ph 6.0) 2,000rpm 10 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 2mL 3mL 100mM 3mL (5 10 >10 / ) 3mL /IPA/NH OH (78:20:2, v/v/v) 4 3mL / / (78:20:2, v/v/v) 1 2mL/min <40 50µL 5µL GC/MS * HPLC 468.4/55.1 -D4* 472.4/ /83.1 -D3* 417.4/ % (50:50, v/v) 0.35mL/min Hypersil GOLD Phenyl 3µm mm SPE-HPLC 30mg 1mL HyperSep Retain- CX ( ) 1mL 1mL 1mL, 20µl 5N HCl 2mL 0.1N HCl, 2 ml 100% 2mL 5% MDA,MDMA 95%-101% Hypersil GOLD aq 5µm,150x4.6mm /95 /20mM K PO,pH mL/min 10µL 214nm

155 /LC/PDA 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM 1mL / 2mL 100mM 3mL 3mL 3mL 100mM <3 1 2mL/min 3mL 3mL 100mM ( >10 5 ) 3mL / (90:10, v/v) 1 2mL/min <40 1,000µL 0.1% ( ) 20µL LC/PDA 0.1% (10:90, v/v) 0.1mL/min ( nm) HPLC Hypersil GOLD 3µm mm delta 9-THC 500mg 6mL HyperSep Aminopropyl ( ) 2mL * 100µL 10 M NaOH / mL ph x 3mL 1 x 3mL 1 x 1mL (ph= 3.0) <3 1 2mL/min 1 x 2mL 1 x 2mL 100mM HCI/ACN (95:5) ( > ) 1 x 200µL 1 x 3mL / (50:50) 1 2mL/min BSTFA <40 50µL 50µL BSTFA 1% TMCS ** / BSTFA 1 2µL GC/MS (TMS ) *** -delta 9-THC-D3* delta 9-THC-D9* delta 9-THC * GC/MS - -delta 9-THC-D9 9-THC-D3 ** TS GC TraceGOLD TG-35MS 30m 0.25m 0.25µm

156 -delta 9-THC (pka 4.5) GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL * 100µL 10 M NaOH / mL ph 3.0( ph ph 3.0) 3mL 3mL 1mL (ph 3.0) <3 1 2mL/min 2mL 2mL 100mM HCl/ (95:5) (5 10 >10 / ) 200µL ( ) 3mL / (50:50) 1 2mL/min BSTFA <40 50µL 50µL BSTFA( 1% TMCS)*** / BSTFA 1 2µL GC/MS (TMS ) *** D 3 - -delta 9 -THC D 3 - -delta 9 -THC* delta 9 -THC * ** ** TS x 1mL GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 200mg 10mL HyperSep Verify-CX ( ) 4mL 1mL ( ) / 5 2,000rpm 10 2mL 100mM (ph =6.0) / ph mM ph 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph 6.0) <3 1 2mL/min 1 x 3mL 1 x 2mL 100mM HCl 1 x 3mL ( >10 5 ) 1 x 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 HPLC HPLC Hypersil GOLD 3µm mm

157 -delta 9-THC Delta-9-THC() Delta-9- THC GC GC/MS 200mg 6mL HyperSep Verify-CX ( ) 1 2mL * / 1mL 2.0mL 100mM ph ( ph ph 3.0) 3mL 3mL 1mL (ph 3.0) <3 1 2mL/min 3mL 2mL 100mM HCl/ (95:5) (5 10 >10 / ) 200µL ( 200µL ) (5 >10 / ) delta-9-thc( ) / 200µL ( 200µL ) 2mL ( delta-9-thc) 3mL / (50:50) 1 2mL/min ( BSTFA ) <40 50µL 50µL BSTFA( 1% TMCS)*** / BSTFA 1 2µL (TMS ) *** D 3 - -delta-9-thc* D 9 - -delta-9-thc* delta-9-thc D 3 - -delta-9-thc* delta-9-thc D 3 -delta-9-thc* Delta-9-THC (303,315,330,343)** * ** delta-9-thc ** TS-38831(10 x 1mL ) GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 156

158 THC 30mg 3mL HyperSep Retain-CX ( ) 2mL * 100µL 10N NaOH / mL ph mL/min 1 x 1mL 1 x 1mL 0.1M HCI/ACN (70:30) ( >10 3 ) 1 x 200µL 2 x 0.5mL / (50:50) 1 2mL/min <40 50µL / 50µL BSTFA 1% TMCS ** / BSTFA 1 2µL GC/MS (TMS ) ( ) -THC-TMS THC-D3-TMS* * -THC-D3-TMS ** TS GC TraceGOLD TG-35MS 30m 0.25m 0.25µm /GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 3.0) * 1mL / 1g (1:4) 2mL 100mM (ph 3.0) / 3mL 3mL 1mL 100mM (ph 3.0) <3 1 2mL/min 4mL 2mL 100mM HCl ( >10 5 ) 3mL 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 100µL 1 2µL ** * D7* * ** GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 157

159 7- GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL * 1mL - - :100mM (ph=5.0) 5,000 F /ml Patella Vulgata / mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 2mL 2mL 20% 100mM (ph= 6.0) ( >10 5 ) 2mL 3mL 2% NH 4 OH 1 2mL/min <40 50µL 50µL MTBSTFA( 1% TBDMCS)*** / MTBSTFA 1 2µL (GC/MS) ** -TBDMS TBDMS D4-TBDMS D4-TBDMS * ** *** TS-48927(10 x 1mL ) GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P / HPLC 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6.0) / 1 5mL 1mL 1g (1:4) / 2mL 100mM (ph 6.0) ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 1mL 100mM 3mL >10 5 2mL CH 4 OH/NH 4 OH (98:2) 1 2mL/min 2.0mL 500µL / 2,000rpm 10 <40 HPLC 158

160 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6) * 1mL / 1g (1:4) 2mL 100mM (ph 6) / 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph 6) <3 1 2mL/min 1 x 3mL 1 x 3mL 100mM HCl 1 x 3mL ( >10 5 ) 1 x 3mL ACN (78:20:2 v/v) 1 2mL/min 50µL 5µL LC/MS MRM 304.2/ D3* 307.2/ / D8* 298.2/ / D8* 326.2/204.2 * -D3 -D8 -D8 HPLC Hypersil GOLD 3µm mm / GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 159 1mL 100mM (ph 6.0) * 1 5mL / 1g (1:4) / 2mL 100mM (ph 6.0) ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 1mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH OH (78:20:2) 4 1 2mL/min IPA/NH OH 4 (ph 11-12) <40 50µL 1 2µL ** D 5 - * * ** GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P

161 / GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6.0) * 2mL / 1g (1:4) / 2mL 100mM (ph 6.0) / ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 2mL 100mM HCl 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 50µL 50µL BSTFA** ( 1% TMCS)*** / BSTFA 1 2µL (GC/MS) ** D 3 - * D 3 - -TMS* TMS * ** ** TS x 1mL GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 160

162 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL 0.5 1g 3mL 100mM (ph 6.0) 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 2mL 100mM HCl 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) 50µL 50µL BSTFA**( 1% TMCS***) / BSTFA 1 2µL (GC/MS) ** D 3 - * D 3 - -TMS* TMS * ** ** TS x 1mL GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 161

163 Delta 9-THC Delta 9-THC -Delta-9-THC 200mg 6mL HyperSep Verify-CX ( ) 1 2mL * 2mL ACN ACN ACN 200µL 2mL (ph~ ) ACN 1mL (84/15/1) /ACN/NH 4 OH 1 x 3mL / / (49:49:2) 1 2mL/min <40 50µL / 50µL BSTFA 1% TMCS *** BSTFA 2µL GC/MS BSTFA (374,346,389) * GC/MS D9- -delta 9-THC D3- - delta 9-THC D3-delta 9-THC ** delta-9 THC *** TS (374,462,477) (380,479,497) GC TraceGOLD TG-35MS 30m 0.25m 0.25µm

164 DHEA GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 5mL * ph ,000rpm 5 3mL 3mL 1mL 100mM (ph 6.0) 3mL > mL 1 2mL/min 2mL 0.2M (ph 7.0) / :1 NaHCO 3 / Na 2 CO 3 ph mL n rpm µL MSTFA***/NH l/ (1000:2:5, 4 v/w/w) rpm 1 GC 1 2µL ** DHEA * * GC/MS 20ng/mL ** *** TS-48910(10 x 1mL ) GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph= 6) * 1mL 2mL 100mM (ph= 6) / ph mM ph / 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph= 6) <3 1 2mL/min 1 x 3mL 1 x 3mL 100mM 1 x 3mL ( >10 5 ) 1 x 3mL / / (78:20:2 v/v) 1 2mL/min <40 200µL 0.1% 5µL LC/MS * 314.2/ /44.1 * 163

165 GC GC/MS 100mg 1mL HyperSep C18 ( : ) 500µL * 500µL20% 3mL 3mL 1 ml 100mM HCI <3 1 2mL/min 3mL 3mL 3mL ( >10 5 ) 1mL2% <40 50µL 50µL BSTFA( 1% MCS)** 50µL MTBSTFA( 1% TMCS)** 50µL µL : ** D10* * ** TS-38831(10 x 1mL ) *** TS-48927(10 x 1mL ) GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P Wolf, C.E. II, Sady J., and Pokalis, A. (1996). Determination of gabapentin in serum using solid phase extraction and gas chromatography. Journal of Analytical Toxicology GC GC/MS 200mg 10mL HyperSep Verify-CX ( : ) 2mL * 1mL - - :100mM (ph 5.0) 5,000 F /ml Patella Vulgata / ,000rpm 10 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 2mL 20% 100mM (ph= 6.0) ( >10 5 ) 2mL 7-3mL 2% NH 4 OH 1 2mL/min <40 50µL 50µL MTBSTFA( 1 % TBDMCS)** / M BS FA 1 2µL : ** D 5 * * ** *** TS-48927(10 x 1mL ) GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 164

166 Gamma-(GHB) 200mg 6mL Retain-AX ( ) GHB 200µg/mL GHB - D6 100µg/mL 10µL 200µL 10µg/mL 25µL 200µL 25µg/mL 50µL 200µL 50µg/mL 100µL 200µL 100µg/mL 200µL QC 1.5mL 25µL * 1mL / N 2 200µL 100mM (ph 6.0) / 15 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph 6.0) Eppendorf 1mL /NH 4 OH (99:1) / 1 N µL 100 L BSTFA( 1% TMCS**) / µL GC/MS * GHB-D6-di-TMS* GHB-di-TMS * GC/MS D6-GHB ** TS GC TraceGOLD TG-35MS 30m 0.25m 0.25µm mg 6mL HyperSep Retain-CX ( ) 1mL 100mM (ph 6) * 1mL 2mL 100mM (ph 6) / 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph 6) <3 1 2mL/min 1 x 3mL 1 x 3mL 100mM 1 x 3mL ( >10 10 ) 1 x 3mL ACN (78:20:2) 1 2mL/min <40 50µL 5µL LC/MS * -D3 MRM PCP 244.3/159.2 PCP-D / / D3* 275.1/131.0 HPLC Hypersil GOLD 3µm mm

167 Gamma- (GBL) Gamma(GHB) 200mg 6mL Retain-AX ( ) 200µL * 100µL 100mM ( ph 6.0 ) / 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph= 6.0) 1 1mL /NH 4 OH (99:1) / N µL 1mL 5 3,000rpm 5 N < µL 100µL BSTFA( 1% TMCS)** / 1 2µL GC/MS * GHB-D6-di-TMS* GHB-di-TMS * GC/MS D6-GHB ** TS GC TraceGOLD TG-35MS 30m 0.25m 0.25µm /GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6.0) * 1 2mL / / 2mL 100mM (ph 6.0) ph mM ph Verify-CX 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL 3mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 100µL 1 2µL * D 4 - * * ** GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 166

168 //LC/MS/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6.0) * 1mL / 2mL 100mM (ph 6.0) / 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL 1mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 100µL 5µL LC/MS/MS * MRM 333.2/ D 5 * 342.3/ / D 5 * 238.3/ % (50:50, v/v) 0.35mL/min HPLC Hypersil GOLD PFP 3µm mm HPLC 200mg 10mL HyperSep Verify-CX ( ) 1mL * 2mL 100mM (ph 6.0) / 2,000rpm 10 ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL 1mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 200µL (1:3) / A 0.1% B:ACN + 0.1% 15 B 30% 50% 1mL/min (254nm) HPLC Hypersil GOLD 5µm mm

169 (LSD) GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 4mL * / 5 2,000rpm 10 2mL 100mM (ph 6.0) / ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL 3mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 20µL 20µL BSTFA**( 1% TMCS)*** / BSTFA 1 2µL * D 3 -LSD-TMS* LSD-TMS * ** ** TS-38831(10 x 1mL ) GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 168

170 (LSD) GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 5mL 2mL 100mM ( ph6.0 ) / ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) <3 1 2mL/min 3mL 3mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 20µL 20µL BSTFA** ( 1% TMCS)*** / BSTFA 1 2µL * D 3 -LSD-TMS* LSD-TMS * ** ** TS x 1mL GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P (SPE - HPLC) 30mg 1mL HyperSep Retain- PEP ( ) 1mL 1mL 1mL, 20µL 1mL 5% 1mL 50:50 HPLC (1) 11-Ketotestosterone (2) (3) (4) Hypersil GOLD 5µm, 150 x 4.6 mm /, 43/57 1 ml / min 10µL 254 nm

171 LSD 200mg 3mL HyperSep Aminopropyl ( ) 1mL 100mM (ph 6) * 1mL / 2mL 100mM (ph 6) / 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph 6) 1 2mL/min 1 x 3mL 1 x 3mL 100mM 1 x 3mL ( >10 5 ) 1 x 3mL ACN (78:20:2 v/v) 1 2mL/min 50µL 5µL GC/MS ACN mL/min :Thermo Scientific TSQ MRM LSD 324.2/223.1 lso-lsd 324.2/281(223.1) Nor-LSD 310.2/209.1 OH-LSD 356.2/338.1 LSD-D3* 327.2/226.1 * LSD-D3 >90% (N=10) 0.1ng/mL HPLC Hypersil GOLD PFP 3µm mm mg 3mL HyperSep Aminopropyl ( ) 1mL 100mM (ph 6) * 1mL / 2mL 100mM (ph 6) / 1 x 3mL 1 x 3mL 1 x 1mL 100mM (ph 6) < 3 1 2mL/min 1 x 3mL 1 x 3mL 100mM 1 x 3mL > x 3mL ACN (78:20:2 v/v) 1 2mL/min 50µL 5µL GC/MS ACN mL/min :Thermo Scientific TSQ MRM 248.2/ D4 * 252.2/ / D4 * 238.1/164.0 * -D4 -D4 >90% (n=10) 10ng/mL HPLC Hypersil GOLD 3µm mm

172 /( ) GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6.0) * 1mL / 1g (1:4) / 5 2mL 100mM (ph 6.0) / ph mM ph 2,000rpm 10 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL 3mL 100mM (ph 4.5) 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 50µL 50µL BSTFA ( 1% TMCS)*** / BSTFA 1 2µL ** - D3 - TMS * TMS D3 - TMS * TMS * ** *** TS-38831(10 x 1mL ) GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 171

173 / /EDDP GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL 100mM (ph 6.0) * 1mL / 1g (1:4) 2mL 100mM (ph 6.0) ph mM ph / 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL 1mL 100mM 3mL ( >10 5 ) 3mL / / (78:20:2, v/v/v) 1 2mL/min <40 100µL 5µL GC/MS * ( ) % 0.1% mL/min MRM 310.2/ D9* 319.2/268.3 EDDP 278.2/234.2 EDDP-D3* 281.4/234.3 HPLC Hypersil GOLD aq 3µm mm

174 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL * 1mL 100mM ( ph 6. 0 ) / ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL 1mL 100mM 3mL ( >10 5 ) 3mL /IPA/NH 4 OH (78:20:2) 1 2mL/min IPA/NH 4 OH (ph 11-12) <40 100µL 1 2µL ** D 9 - * * ** GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P / GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL / * 1mL 100mM (ph 6.0) / 2mL 100mM (ph 6.0) / ph mM ph 3mL 3mL 1mL 100mM (ph 6.0) < 3 1 2mL/min 3mL ( >10 5 ) 2mL 3mL / (50:50) <40 100µL 1 2µL ** * D7 * * ** GC TRACE TR-DoA5 15m 0.25mm 0.25µm TRACE TR-DoA35 15m 0.20mm 0.33µm 26AF130P 26AC497P 173

175 / LC/MS 200mg 10mL HyperSep Verify-CX ( ) 1mL100mm (PH 6.0) * 1mL / 2mL 100mm (PH 6.0) Verify-CX 3mL 3mL 1mL100mm (PH 6.0) : <3 1 2mL/min 3mL ( >10 5 ) 2mL 3mL / (50:50) 1 2mL/min <40 100µL 5µL GC/MS : MRM 251.2/ D7* 258.2/138.2 * : 0.1 % (30:70,v/v) :0.35mL/min GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL * 2mL100mm (ph 6.0) ph mm ph / Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 ZmL/min 3mL 2mL200mm HCI ( >10 5 2mL 3mL ( >10 5 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/NH 4 0H (ph 11-12) <40 100µL 1 2µL (GC/MS) ** D4* D3* * D,- D 4 - ** GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm 26AF130P TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AC497P 174

176 GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 2mL 400µL HCI 200µL 10% / ,000rpm µL 50% / 50% ph 5-6 2mL / 60 ( ) 200µL 10% ph 5-6 2,000rpm 10 Verify-CX 3mL 3mL 1mL 100mm (ph 6.0) : <3 1 2mL/min 3mL 3mL 100mm (ph 4.5) 3mL ( >10 5 ) 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/N H 4 0H (ph 11-12) <40 100µL 100µL BSTFA ( 1%TMCS)*** / : BSTFA 1 2µL : ** D 4 - * D 4 - TMS* TMS* TMS TMS N- TMS TMS D 3 - TMS* D 6 - TMS* TMS TMS TMS D 3 - TMS* D 6 - TMS* TMS TMS D 3 - TMS D 6 - TMS* TMS D 3 - TMS TMS D 3 - TMS D 6 - TMS* TMS D 3 - TMS TMS * GC/MS D 6 - D 6 - L0D/L0Q ** *** TS38831(10x 1mL ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P 175

177 - GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) - 2mL 400µL HCI 200µL 10% / ,000rpm µL 50% / 50% ph mL * 1mL - ( F /ml) ,000rpm M Na0H ph 5-6 Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 3mL100mm (ph 4.5) 3mL ( >10 5 ) 3mL /IPA/NH 4 0H (84:12:4) 1 2mL/min : ; IPA/NH 4 0 H (ph 11-12) <40 200µL 1:1 : / <40 50µL / (70:30, v/v) 1 2µL : ** D3* D3* * GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm 26AF130P TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AC497P 176

178 / LC/MS/MS 200mg 10mL HyperSep Verify-CX ( : ) 1mL 100mm (ph 6.0) * 1mL / 2mL 100mm (ph 6.0) Verify-CX 3mL 3mL 3mL 100mm (ph 6.0) : <3 1 2mL/min 3mL 3mL 100mm 3mL ( >10 5 ) 3mL / / (78:20:2,v/v/v) 1 2mL/min <40 100µL 5µL GC/MS/MS : MRM 330.0/ D6* 336.0/76.1 * : ( ) % % 0.1% mL/min HPLC Hypersil GOLD Phenyl 3µm, 150 x 2.1 mm mg 3mL HyperSep Retain-CX ( : ) 1mL 1mL 100mM (ph 6.0) / HyperSep Retain-CX 1 ZmL/min 1x1mL 1 x 1 ml 100mm 1x1mL ( >10 3 ) 2 x 0.5mL /IPA/NH 4 0H(78/20/2) 1 ZmL/min 1 1% <40 100µL 1 2µL GC/MS : ( ) D5* * -D5 GC TraceGOLD TG-5SiIMS 30m x 0.25mm x 0.25µm

179 / GC GC/MS 200mg 10mL HyperSep Verify-CX ( : ) 1mL 100mm (ph 6.0) * 1mL / 1g (1:4) / ph mm ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 3mL 100mm 3mL ( >10 5 ) 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/NH 4 0H (ph 11-12) <40 100µL 1 2µL. : ** D5- * * ** GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P / GC/MS 200mg 10mL HyperSep Verify-CX ( : ) 1mL 100mm (ph 6.0) * 1mL / 2mL 100mm (ph 6.0) / ph mm ph / Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 3mL 100mm 3mL ( >10 5 ) 3mL / / (78:20:2,v/v/v) 1 2mL/min : <40 100µL 5µL GC/MS/MS : MRM 330.0/ D6* 336.0/76.1 * : /0.1% (33:67,v/v) :0.35mL/min : HPLC Hypersil GOLD 3µm, 150 x 2.1 mm

180 / GC GC/MS 200mg 10mL HyperSep Verify-CX ( : ) 1mL 100mm (ph 6.0) * 1mL / 2mL 1g (1 :4) / p mM ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 3mL 100mm 3mL ( >10 5 ) 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; }PA/N H40 H (ph 11-12/ <40 100µL 1 2µL : ** D5- * * ** : ( ) 1 35% ph 6 SP ( ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P / LC/PDA 200mg 10mL HyperSep Verify-CX ( : ) 1mL 100mm (ph 6.0) * 1mL / 1g(1:4) 2mL100mm (ph 6.0) ph mm ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 3mL 100mm 3mL ( >10 5 ) 3mL ( >10 5 ) 3mL / / (78:20:2, v/v/v) 1 2mL/min : <40 100µL 0.1% ( ) 50µL LC/MS/MS : * * :1 ml/min : : 0.1% (25:75) : (250nm) HPLC Hypersil GOLD 3µm, 150 x 4.6 mm

181 / LC/MS/MS 200mg 10mL HyperSep Verify-CX ( : ) 1mL 100mm (ph 6. ) 1mL / 1g}1:4) 2mL 100mM (ph 6.0) Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/mi 3mL 3mL 100mm 3mL ( >>1 5 ) / 3mL / / (78:20:2,v/v/v) 3mL /IPA/NH 4 0H(78:20:2,v/v/v) 1 2mL/mi : ; IPA/NH 4 0H (ph11-12) <400C 100µL 0.1% / <40 100µL 5µL LC/MS/MS : MRM 340.0/58.0 -D11* 351.2/ / D5* 331.0/ mL/min : ( ) % % 0.1% HPLC Hypersil GOLD Phenyl 3µm, 50 x 2.1 mm (SPE-LC/MS ) 30mg 3mL HyperSep Retain-CX ( : ) : 1mL 1mL 1 ml 30mm : 1mL 0.5% : 1mL 1mL : 1mL5% LC/MS : Hypersil GOLD 3µm, 50 x 2.1 mm : : 100mm : =70:30 : 200µL/min MS : (ESI) (SRM) 4500V 350 : : 97-99% 180

182 -4- GC GC/MS 200mg 10mL HyperSep Verify-CX ( ) 5mL *( -4- -D10-TMS) 2mL 100mm (ph 6.0) / 12,500 25,000 b- / ,000rpm 10 ( ) SPE Verify-CX 3mL 3mL 1mL100mm (ph 6.D) : <3 1 2mL/min 3mL 2mL 20% 1 ml 100mm ( >10 3 ) 2mL 3mL / (50:50) 3mL ( >10 3 ) -4-3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; }PA/N QOH (ph 11-12) <35 5µL / 5µL MSTFA*** : MSTFA 1 2µL : ** -4- -TMS (291) -4- -D10-TMS* (301) * ** TS ( 10x1mL ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P Greishaber A. Moore, K. Levine, B, and Smith M.(1999, Novembe). The detection of psilocin in human urine. Presented at the TRI Services Meeting 181

183 ( ) 200mg 10mL HyperSep Verify-CX ( ) 1mL 4mL (ph 5 7) / 5 2,000rpm 10 2mL 100mm (ph 6.0) / ph mm ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <c3 1 2mL/min 3mL 3mL ( >10 5 ) 2mL 3mL / (50:50) <5mL/min 3mL ( >10 5 ) 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/NH 4 0H (ph 11-12) <40 100µL 90µL ( 1.0mL) H PLC 200mg 10mL HyperSep Verify-CX ( : ) 4mL 2mL 100mm (ph 6.0) 1mL / / 2,000rpm 10 ph mm ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 1 ml 100mm 3mL ( >10 5 ) 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/N H 4 0H (ph 11-12) <40 200µL : (1:3) / 30 10µL LC 235nm : 30% 0.25 M (ph 2.7) :2mL/min HPLC Hypersil GOLD C8 3µm, 150 x 4.6 mm

184 -4-200mg 10mL HyperSep Verify-CX ( ) 1 ml 100mm (ph 6) 1mL 2mL 100mm (ph 6) / 1mL 1mL (p :100mm (ph=5.0) F /ml Patella Vulgata) / ,000rpm 10 HyperSep Verify-CX 1x3mL 1x3mL 1 x 1 ml 100mm (ph 6) : <3 1 2mL/min 1x3mL 1 x 3mL 100mm 1x3mL ( >10 5 ) 1x3mL ;ACN; (78:20:2 v/v) 1 2mL/min 5µL 5µL GC/MS : %ACN 0.1% mL/min :Thermo Scientific TSQ MRM / D10* 215.2/68.2 * -4- -D10 >90% (n=10) :10ng/mL HPLC Hypersil GOLD PFP 3µm, 100 x 2.1 mm (SPE-HPLC) 200mg 10mL HyperSep Verify-CX ( ) : 1mL 1mL2% : 1mL 2% : 1mL2% 1mL : 1mL 5% : : 72%-98% HPLC : Hypersil GOLD 5µL, 150 x 4.6mm : : A.0.05M ph3; B: (85:15) : 1 ml/min t: 10µL UV; 210nm :

185 / GC GC/MS PFPA (PFAA) - 200mg 10mL HyperSep Verify-CX ( : ) 1mL100mm (ph 6.0) * 1mL / 1g(1:4) / ph mm ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 1 ml 100mm 3mL ( >10 5 ) SMA 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/NH 4 0H (ph 11-12) 30µL DMF*** <40 30µL PFPA (PFAA) - 50µL PFPA(PFAA)**** 50µL PFPOHt <40 100µL 1 2µL : ** D 5 - * D 5 - * * GC/MS D 5 - D 5 - ** *** TS-20672(50mL ) *** TS-65193(10x1mL ) *** TS-65195(10x1mL ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P 184

186 / GC GC/MS TMS - 200mg 10mL HyperSep Verify-CX ( : ) 2mL * 1 ml 100mm (ph 6.0) / ph mm ph Verify-CX 3mL 3mL 1mL 100mm (ph 6.0) : <3 1 2mL/min 3mL 1 ml 100mm 3mL ( >10 5 ) SMA 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/N H 4 0H (ph 11-12) 3µL DMF <40 30µL TMS - 50µL BSTFA( 1 % TMCS) 5µL µL : ** D 5 - * D 6 - * D 10 - * D 11 - * D 5 - * D 8 - * D 9 - * * GC/MS ** *** TS-20672(50mL ) *** TS-38831(10x1mL ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm 26AF130P TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AC497P 185

187 / GC GC/MS - 4-CB(4- ) 200mg 10mL HyperSep Verify-CX ( : ) 2mL * 1 ml 100mm (ph 6.0) / ph mM ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <c3 1 2mL/min 3mL 1 ml 100mm 3mL ( >>10 5 ) SMA 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/NH 4 0H (PH11-12) 30µL DMF*** <40 30µL 4-CB(4- ) - 20µL 4-CB 00µL µL : (4-CB) ** D 5 - * D 5 - * D 5 - * D 5 - * D 6 - * * ** *** TS-20672(50mL ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P (cs), (CN) ((0C) GC/MS 200mg 10mL HyperSep Verify-CX ( : ) Kimwipe Verify-CX 3mL 3mL 1 ml 100mM (ph 6.0) : : <3 <3 1 ml/mi 3mL 3mL ( >10 5 ) 1mL <40 200µL / GC/MS

188 / / GC GC/MS 200mg 10mL HyperSep Verify-CX ( : ) : 2mL 1 ml 100mm (ph 6.0) / ph mm ph : 1mL * 1mL (ph 6.0) 4mL / 5 2,000rpm 10 2mL 100mm (ph 6.0) / ph mm ph Verify-CX 3mL 3mL 1 ml 100mm (ph 6.0) : <3 1 2mL/min 3mL 1mL100mm ( >10 5 ) 2mL ( 1 ) 3mL / (50:50) <2mL/min <40 100µL 1 2µL 3mL ( >10 5 ) ( 2 ) 3mL /IPA/NH 4 0H (78:20:2) 1 2mL/min : ; IPA/NH 4 0H (PH 11-12) <40 100µL 1 2µL GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm 26AF130P TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AC497P : (1) 1 ( ) 2 ( ) (2) ( DMF) ; 30 50µm DMF( 2 ) (3) 1% 2 100µm 1 187

189 60mg 6mL HyperSep Retain-CX ( ) µL µL 100mm (ph= 6.0) / 10 PH mm ph HyperSep Retain-CX 1 x 200µL 1x200µL 1 x 200µL 100mm (ph=6.0) 1 ml/min 1 x 500µL 1 x 500µL 100mm 1 x 500µL ( >10 5 ) 1 x 800µL /IPA/NH 4 0H (70:26:4) 1 ml/min : ; IPA/NH 4 0H (ph 11-12) PCP 5 100µL 5% 5 <40 *: 50µL PFPA(PFAA) / <40 50µL : 200µL 1:1 / / < µL * GC/MS 50mg 1mL HyperSep Verify-CX ( : ) µL µL 100mm (ph 6.0) / ph mm ph Verify-CX 200µL 200µL 200µL 100mm (ph 6.0) 1mL/min ( ) 500µL 500µL 100mm 500µL ( > µL /IPA/NH 4 0H (70:26:4) 1mL/min ( ) : ; IPA/N H 4 0H (ph 11-12) PCP 5 10µL 5% (5 ) <40 *: 50µL PFPA(PFAA)** <40 50µL * 200µL 1:1 PFPA(PFAA)** (10x1mL TS-65193) µL 1 µl * ** TS-65193(10X1ML ) : GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P 188

190 60mg 6mL HyperSep Retain-CX ( ) µL * µL 100mm (ph=6.0) / 10 ph mm ph HyperSep Retain-CX 1 x 200µL 1 x 200µL 1 x 200µL 100mm HCI 1mL/min 1 x 500µL 1 x 500µL 100mm 1 x 500µL / (50:50) ( >10 5 ) 1 x 800µL /IPA/NH 4 0H (70:26:4) 1mL/min : ; IPA/NH 4 0H (ph 11-12) <40 : 100µL PFPA (PFAA) HFIP <40 50µL TMS: 25µL BSTFA( 1 % TMCS)** 25µL / : BSTFA 2µL GC/MS : MRM ,303 -D3* ,306 -TMS ,361 -D8-TMS* ,369 * -D3 -D8-TMS ** TS-3B831 HPLC Hypersil GOLD TG-5MS 30mx0.25m x0.25µm

191 / 200mg 10mL HyperSep Verify-CX ( : ) ng / 100µL / µL (ph 6) * 1 100µL 10 3µL 100mm (ph 6) / HyperSep Verify-CX 1x3mL 1x3mL 1 x 1µL 100mm (ph 6) : <3 1 2mL/min 1x3mL 1x3mL1% 1x3mL ( >10 5 ) 1x3mL ACN; (78:20:2 v ) 1 2mL/min <<400C 2 L 5 LC/MS MRM 333.2/ D5* 342.2/ /84.1 -D5* 238.3/84.1 * -D5 -D5* HPLC Hypersil GOLD aq 5µm, 50 x 3mm (THC) GC/MS 200mg 10mL HyperSep Verify-CX ( : ) 1mL 50ng/mL (THCA D9-TMS) mL 10 Verify-CX 3mL 3mL 1mL 0.1N HCI 1 ml/min ( ) 2mL 2mL 0.1N HCI/ (70:30) ( >10 5 ) 200µL THC 2mL / (50:50) 1 ml/min ( ) <40 50µL MSTFA*** µL 1 2µL : ** THCA-TMS THCA D9-TMS* * ** *** TS ( 10x1mL ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P Janet Putnam, Assistant Laboratory DirectorlRP Advanced Toxicology Network, Memphis, TN 190

192 GC/MS 50mg 1 ml HyperSep Verify-CX ( ) µL 800µL 100mm (ph 6.0) / 10 ( ph ) 100mm ph Verify-CX 200µL 200µL 200µL 0.1N HCI 1 ml/min ( ) 500µL 500µL 100mm HCI 500µL / (50:50) ( >10 5 ) 800µL /IPA/NH 4 0H (70:26:4) 1mL/min ( ) : ; IPA/NH 4 0H (ph 11-12) <40 : 100µL PFPA (PFAA) HFIP <40 50µL TMS: 25µL BSTFA( 1 % TMCS)** 25µL / : BSTFA 1 2µL : ( ) D3* D8-TMS* * ** TS x1mL *** TS x1mL GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P 191

193 (THC) GC/MS 50mg 1mL HyperSep Verify-CX ( : ) µL * mL / 10 Verify-CX 200µL 200µL 200µL 100mm HCI 1mL/min ( ) 500µL 500µL 0.2N HCI 500µL 100mm HCI/ (70:30) ( >10 5 ) THC 800µL / (75:25) 1mL/min ( ) <40 25µL BSTFA( 1 % TMCS}*** 25µL : BS FA 1 2µL : ** THCA-TMS THCA D9-TMS* THC * ** *** TS-38831(10x 1mL ) GC TRACE TR-DoA5 15m x 0.25mm x 0.25µm TRACE TR-DoA35 15m x 0.20mm x 0.33µm 26AF130P 26AC497P THC 60mg 6mL HyperSep Retain-CX ( : ) µL * µL / 10 Verify-CX 1x200µL 1x200µL 1x200µL 100mm HCI 1mL/min 1x500µL 1x500µL 0.2N HCI 1x500µL 100mm HCI/ACN(70:30) ( >10 5 ) 1X800µL : (25:75) 25µL BSTFA( 1 % TMCS}** 25µL / : BSTFA 2µL GC/MS : ** THCA-TMS THCA D3-TMS* * THC-D3-TMS ** TS GC Trace GOLD TG-35MS 30m x 0.25m x 0.25µm mL/min <40 192

194 SOLA SPE 10mg/1 ml 10mg/2mL 96 SOLA SPE (ml) SOLA HRP 10mg 1mL / SOLA SCX 10mg 1mL / SOLA SAX 10mg 1mL / SOLA 96 (ml) SOLA HRP 10mg 2mL / SOLA SCX 10mg 2mL / SOLA SAX 10mg 2mL / SOLA SPE 1 SOLA SPE SOLA SPE 2 SOLA 30 SPE (i) SOLA. 3 SOLA 193

195 HyperSep SPE Thermo Scientific SPE HyperSepSPE 96- HyperSep SPE HyperSep MEPS 1mL 75mL 25mg 10g SPE 3.6µL HyperSep-96 HyperSep SPE 96 HPLC HPLC HyperSep Hypercarb HyperSep Hypercarb SPE 25mg 1mL / 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / HyperSep Hypercarb SPE 50mg 1mL / 100mg 1mL / 200mg 1mL / 500mg 1mL g 1mL g 1mL HyperSep Retain PEP HyperSep Retain PEP SPE 30mg 1mL / 30mg 3mL / 60mg 3mL / 60mg 6mL / 100mg 6mL / 150mg 6mL / 200mg 3mL / 200mg 6mL / 500mg 3mL / 500mg 6mL / 1g 25mL / 2g 25mL / HyperSep-96Retain PEP 5mg 1mL / 10mg 1mL mg 1mL / 60mg 1mL / 5mg 1mL mg 1mL mg 1mL mg 1mL

196 HyperSep Retain-CX HyperSep Retain-CX SPE 30mg 1mL / 30mg 3mL / 60mg 3mL / 60mg 6mL / 100mg 6mL / 150mg 6mL / 200mg 3mL / 200mg 6mL / 500mg 3mL / 500mg 6mL / 1g 25mL / 2g 25mL / HyperSep-96Retain PEP 5mg 1mL / 10mg 1mL / 30mg 1mL / 60mg 1mL / 5mg 1mL mg 1mL mg 1mL mg 1mL HyperSep Retain-AX HyperSep Retain-AX SPE 30mg 1mL / 30mg 3mL / 60mg 3mL / 60mg 6mL / 100mg 6mL / 150mg 6mL / 200mg 3mL / 200mg 6mL / 500mg 3mL / 500mg 6mL / 1g 25mL / 2g 25mL / HyperSep-96Retain-AX 5mg 1mL / 10mg 1mL / 30mg 1mL / 60mg 1mL / 5mg 1mL mg 1mL mg 1mL mg 1mL HyperSep C18 HyperSep Silica C18 SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 C18 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL

197 HyperSep C8 HyperSep C8 SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 C18 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL HyperSep Phenyl HyperSep Phenyl SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 Phenyl 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL HyperSep Silica HyperSep Silica SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 Silica 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL

198 HyperSep SAX HyperSep SAX SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 SAX 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL HyperSep SCX HyperSep SCX SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 SCX 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL HyperSep Verify AX HyperSep Verify-AX SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 Verify-AX 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL

199 HyperSep Verify-CX HyperSep Verify-CX SPE 130mg 1mL / 200mg 6mL / 300mg 3mL / 500mg 3mL / 500mg 6mL / 1g 10mL / 2g 6mL / HyperSep-96 Verify-CX 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL HyperSep Florisil HyperSep Florisil SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 Florisil 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL HyperSep Aminopropyl HyperSep Aminopropyl SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 Aminopropyl 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL

200 HyperSep Cyano HyperSep Cyano SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 Cyano 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL HyperSep Diol HyperSep Diol SPE 50mg 1mL / 100mg 1mL / 200mg 3mL / 500mg 3mL / 500mg 6mL / 1g 6mL / 2g 15mL / 5g 25mL / 10g 75mL / HyperSep-96 Diol 10mg 1mL / 25mg 1mL / 50mg 1mL / 100mg 1mL / 10mg 1mL mg 1mL mg 1mL mg 1mL

201 HyperSep SPE HyperSep Javelin SPE Retain PEP Retain-CX Retain-AX Hypercarb 2.1mm 10mm / 3.0mm 10mm / HyperSep UNIGUARD SPE Retain PEP Retain-CX Retain-AX Hypercarb 2.1mm 10mm / 3.0mm 10mm / SPE HyperSep HPLC Retain PEP Retain-CX Retain-AX Hypercarb 2.1mm 20mm / 3.0mm 20mm / HyperSep HyperSep SPE 96- / / / / HyperSep-96 HyperSep HyperSep HyperSep HyperSep / 1mL / 2mL / 1mL 3mL 6mL SPE / 1mL / 200

202 HyperSep 16 Corian0 16 Teflon0 Luer Corian0 24 Teflon0 Luer / / / Luer / / Luer Lock / 201

203 HyperSep SLE 96 / (SLE) SLE 200mL, SLE : LLE LLE LL LLE : (1) SLE 5-10 (2) (3) HyperSep SLE 96 HyperSep SLE SLE(pH=7) 200mg mg mg g g g g HyperSep SLE SLE(pH=9) 200mg mg mg g g g g HyperSep SLE 96 SLE(pH=7) 200mg W 1/ 300mg W 1/ 400mg W 1/ 500mg W 1/ HyperSep SLE SLE(pH=9) 200mg W 1/ 300mg W 1/ 400mg W 1/ 500mg W 1/ 202

204 HyperSep SPE -QuEChERS Thermo Scientific QuEChERS( ) SPE QuEChERS HyperSep SPE -QuEChERS QuEChERS 6g MgSO 4, 1.5g 50mL / 4g MgSO 4, 1g 50mL / 1.5g 1.5g 750mg 50mL / 4g 1g 1g 500 mg 50mL / QuEChERS 6g 1.5g 50mL mL / 4g 1g NaCI 50mL mL / 6g 1.5g 1.5g 750mg 50mL mL / 4g 1g 1g 500mg 50mL mL / 900mg MgSO 4 300mg PSA 150mg 15mL 15mL / 900mg MgSO 4 300mg PSA 150mg C18 15mL 15mL / 750mg MgSO 4 250mg PSA, 250mg C mL 15mL / 900mg MgSO 4, 300mg PSA 15mL 15mL / 900mg MgSO 4 150mg PSA 15mL 15mL / 900mg MgSO 4 150mg PSA 45mg 15mL 15mL / 900mg MgSO 4 150mg PSA 15mg 15mL 15mL / 1200mg MgSO 4 400mg PSA 15mL 15mL / 1200mg MgSO 4 400mg PSA 400mg C18 15mL 15mL / 1200mg MgSO 4 400mg PSA, 400mg C18 400mg 15mL / 900mg MgSO 4 150mg PSA 150mg C18 15mL / 150mg MgSO 4 50mg PSA 50mg 15mL / 150mg MgSO 4 300mg PSA 150mg Chlorofiltr 15mL / 900mg MgSO 4 300mg PSA 150mg 15mL / 900mg MgSO 4 300mg PSA 150mg C18 15mL / 150mg MgSO 4 50mg PSA, 50mg 15mL / 900mg MgSO 4 300mg PSA 15mL / 900mg MgSO 4 150mg PSA 15mL / 900mg MgSO 4 50mg PSA 45mg 15mL / 900mg MgSO 4 150mg PSA 15mg 15mL / 1200mg MgSO 4 400mg P SA 15mL / 1200mg MgSO 4 400mg PSA 400mg C18 15mL / 1200mg MgSO 4 400mg PSA, 400mg C18 400mg 15mL / 203

205 QuEChERS 900mg MgSO 4 150mg PSA 150mg C18 15mL / 150mg MgSO 4 10mg PSA 50mg 15mL / 150mg MgSO 4 300mg PSA 150mg Chlorofiltr 15mL / 400mg PSA 200mg 6mL / 500mg PSA 250mg 6mL / 500mg PSA 500mg 6mL / 400mg PSA 200mg 6mL / 500mg PSA 250mg 6mL / 500mg PSA 500mg 6mL / 150mg MgSO 4 50mg PSA 50mg 2mL / 150mg MgSO 4 50mg PSA 2mL / 150mg MgSO 4 50mg PSA 50mg C18 2mL / 150mg MgSO 4 25mg PSA 2mL / 150mg MgSO 4 25mg PSA 25mg C18 2mL / 150mg MgSO 4 25mg PSA 2.5mg 2mL / 150mg MgSO 4 25mg PSA 7.5mg 2mL / 150mg MgSO 4 50mg PSA 50mg C18 50mg 2mL / 150mg MgSO 4 50mg PSA 50mg 2mL / 150mg MgSO 4 50mg PSA 2mL / 150mg MgSO 4 50mg PSA 50mg C18 2mL / 150mg MgSO 4 25mg PSA 2mL / 50mg MgSO 4 25mg PSA 25mg C18 2mL / 150mg MgSO 4 25mg PSA 2.5mg 2mL 2mL / 150mg MgSO 4 25mg PSA 7.5mg 2mL / 150mg MgSO 4 50mg PSA, 50mg C18 50mg 2mL / HyperSep SPE Mylar Mylar 4000mg 1000mg / Mylar 6000mg 1500mg ( ) / Mylar 6000mg 1500mg / Mylar 6000mg 1500mg 1500mg 750mg / Mylar 4000mg 1000mg mg / Mylar 1.2g / Mylar 4g 0.5g / HyperSep SPE mg 100mg / 4000mg 100mg 500mg 1000mg / 4000mg 1g / 6000mg 1500mg / 6000mg 1500mg / 6000mg 1500mg 1500mg 1750mg / 8000mg 200mg / 8000mg 3500mg / 204

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