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3 ) ) Biological m onitoring ) ) mod el ) (Method valid ation) (1) (LOD) (LOQ) (2) (Reproducibility test) (3) (Stand ard calibration cu rve lin erity) (4) (Storage stability test) (5) (Breakthrou gh test) (6) (Trapping efficiency test) (Recovery test) (1) (2) (Field recovery test) (3) (4) surfactant(sur-ten ) (5) (acetone)
4 (1) (2) (3) (4) (5) ) Biological m onitoring (1),, (2), (3) Methid athion Methid athion Methid athion (4) Biological m on itorin g I Methid ath ion
5 Methid athion methid athion methid athion methid athion methid athion (LOD) (LOQ) Fu rathiocarb Fu rathiocarb furathiocarb furathiocarb furathiocarb furathiocarb
6 (5) : Methid ath ion Methid ath ion (6) Biological m on itorin g II - Field test Meth id athion Methid athion methid athion methid athion Fu rathiocarb
7 Fu rathiocarb furathiocarb fruathiocarb (7) : Biological m onitoring Methid athion ChE Furathiocarb ChE
8 %, 77%..,,. (REI; Reentry interval)..,,,.. (89 93 ) 3,202,0004, %, 1/ , ,282 M/ T
9 ,,.,. (EPA) ,453ha.,, 96 (, )37,848 ha 13.0%(4,352 ha).,,,,, 90 23,698 ha 34 ha. ().,,,,,, ( 15 ) - 2 -
10 .,,..,,,.. 2,,..,.,,,,, biological m onitoring, m anu al, mod el., p atch,, DDVP, bifenthrin., PBPK mod el EPA
11
12 1. 1, 2biological m onitoring,, m ethid athion, furathioncarb, mod el. (3 ) biological m onitoring, mod el 3,. 1) 1, 2 (, ), mod el,,.,,, biological sam ple(, ),,. 2) Biological monitoring 2 methid athion, furathiocarb,
13 ,. 3) biological m onitoring methid athion furathiocarb. 4) model.. (toxicokinetics).,,,.. (Physiologically-based Pharmacokinetics, PBPK), (And ersen et al., 1987). PBPK (PBPK Mod el),,, (Kenyon et al., 1995).,., PBPK (Sp ecies Extrap olation)
14 1). (Method validation) (1) (LOD ) (LOQ) (LOD) (LOQ) ( 1). 1. Pes ticide LOD(ng ) LOQ(ng) Methidathion F urathiocarb (2) (Reproducibility test) CV %, ( 2). 2. pesticide Methidathion Furathiocarb Concentra tion(ppm) Height Averag e CV(% ) SD (3) (Standard calibration curve linearity) lin earity 4 ( 3)
15 3. Pesticide Calibration curv e 1 3 Methidathion y=3.2515x , r= y=2.3719x , r= y=1.9698x , r= Furathiocarb y=2.4307x , r= y=6.7227x , r= y=5.7197x , r= (4) (Storage stability test). 7. ( 4, 5, 6, 7) 4. Patch Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%) Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%)
16 6. Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%) XAD Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%.) (5) (Breakthrough test) 2. ( 8) 8. (ug) Methidathion 152 F urathiocarb 50 (ug ) CV 1 2 (%) (%) T rap T rap ND ND ND ND ND ND 93.2 (6) (Trapping efficiency test) - 9 -
17 XAD. ( 9) 9. (ug) (ug) CV (% ) (%) (%) Methidathion 500 F urathiocarb (Recovery test) (1) (Matrix extraction efficiency, Recovery test). Fu rathiocarb 75.4%, 110%.,. ( 10, 11, 12, 13) 10. Patch Methidathion Furathiocarb (ug ) (ug ) % CV Recovery (%) LOQ LOQ LOQ LOQ LOQ LOQ
18 Methidathion Furathiocarb (ug ) (ug ) % CV Recovery (%) LOQ LOQ LOQ LOQ LOQ LOQ Methidathion Furathiocarb (ug ) (ug ) % CV Recovery (%) LOQ LOQ LOQ LOQ LOQ LOQ XAD Methidathion Furathiocarb (ug ) (ug ) % CV Recovery (%) LOQ LOQ LOQ LOQ LOQ LOQ (2) (Field recovery test)
19 . Meth id ath ion XAD ( 14). 14. (ug ) Methidathion Furathiocarb (ug) (% ) Patch XAD Patch XAD (3) (Soil Recovery test) 50 g100 m l acetone 60 rotary sh aker 250 rp m. 2 Whatm an N o. 2 filter. acetone, 100 m l, 100 m l Eth yl Acetate 2. Eth yl Acetate gas chrom atograp h. 50 gmethid ation 12.5 u g, 125 u g, 2500 u g Fu rathiocarb 2.5u g, 12.5u g, 125u g. Meth id ath ion Fu rath iocarb 80%, 90%. ( 15)
20 15. Spiked level(ug) Methidathion (ug ) (%) Mean S.D. Spiked lev el(ug ) Furathiocarb (ug ) (%) Control - ND ND Mean S.D (4) surfactant(sur-ten ). 40 (405 cm 2) 0.02% su rfactant 70 m l 30 rotary shaker 2 (200 rp m, 2 ). Glassw ool 70 m l Ethyl acetate 2, Ethyl acetate(5 20m l) gas chrom atograp h. su rfactant 1 u g, 10 u g, 100 u g Meth id ath ion Fu rathiocarb % su rfactant aerosol OT 75% su rfactan t(sod iu m d ioctyl su lfosu ccinate in m ixtu re of ethanol and w ater). su rfactant(sur-ten ) Meth id ath ion 101%, 118%, 103%(LOQ, 10 LOQ, 100 LOQ), 107%. Fu rathiocarb 93%, 112%, 105%(LOQ, 10 LOQ, 100 LOQ), 103% ( 16)
21 16. su rfactan t(sur-ten ) Spiked level(ug ) Methidation (ug) (%) Mean S.D. (ug) Furathiocarb (ug) (%) Control - ND* ND** - - LOQ LOQ LOQ Mean S.D * N D : N on-d etectable( < 0.05 u g ) - Meth id ath ion ** N D : N on-d etectable( < 0.05 u g ) - Fu rathiocarb (5) (acetone) su rfactant 80 m l acetoneacetone m l Waring blen d er1 3, 100 m l aceton e,. acetone hexane(5 10 m l) sep -p ak colum n, 10 ml (hexan e/ acetone=9/ 1) 10 m l gas chrom atograp h. 1 u g(1 p p m, 1 m l), 10 u g(10 p p m, 1 m l), 100 u g(100 p p m, 1 m l)3. Su rfactant(sur-ten ) su rfactant. Su rfactant acetone, su rfactant ( 17)
22 17. (aceton e) Spiked level(ug ) Methidation (ug) (%) Mean S.D. Spiked level(ug ) Furathiocarb (ug) (%) Control - ND* ND** - - LOQ LOQ LOQ Mean S.D * N D : N on-d etectable( < 0.05 u g ) - Meth id ath ion ** N D : N on-d etectable( < 0.05 u g ) - Fu rathiocarb. 1, 2 m ethid athion fu rath iocarb. ( 1 )1, 2, 3, 5, 7 ( 4 ),, 1, 2, 3, 5, 7,. (1) Methid athion, fu rathiocarb.. (1, 2,) 2m eth id ath ion 1086u g/ hr, fu rath iocarb 181 u g/ hr
23 1. Methidathion D ermal exposure 2. Furathiocarb D ermal exposure
24 (2). (3),. m u lching. (4) 1methid athion 222.6u g/ cm 2, furathiocarb 35.3ug/ cm 2, u g/ cm2, 2.6u g/ cm2. (3, 4).. su rfactant acetone m ethid athion ug/ cm2, u g/ cm2, 3 8.3u g/ cm 2, 5 2.1ug/ cm 2, 7 u g/ cm2, furathiocarb ug/ cm 2, 2 7.4ug/ cm 2, 3 4.3ug/ cm 2, 5 1.9ug/ cm 2, 7 1.1ug/ cm
25 3. Methid athion d islod geable resid ue d issipation. 4. Fu rathiocarb d islod geable resid u e d issip ation
26 (5). TF = D Eh /D FR, or D Eh = TF D FR AD D = (D Eh ABS H/D )/BW = (TF D FR ABS H/D )/BW AD D SL = [(TF ABS H/D )/BW] D FRSL, or D FRSL = (AD D SL BW)/(TF ABS H/D ) ln[d FRSL] = ln[d FR0] + K T T = {ln[(ad D SL BW)/(TF ABS H/D )] - ln[d FR0]} K-1 ADD = absorbed daily dosage DEh = hourly dermal exposure ABS = dermal absorption H/D = hours w orked per day TF = transfer factor SL = safe level BW = body w eight D FR= dislogdeable foliar residues, -M ethid ath ion Derm al Exp osu re stu d y DAT1 : u g/ hr DAT2 : u g/ hr Calcu lation of reentry in terval DFR0 = 223 ng/ cm 2 K = ng/ cm 2/ d ay TF = 4872 cm 2/ hr Available d ata ADDSL = 1000 ng/ kg/ d ay (=ADI) BW = 60 kg ABS = % So. Reentry Interval T=[ln{( )/( )}-ln(223)] ( )-1 =1.2 day ( 28 hours )
27 Methid ath ion EPA REI 48(from Toxicological d ata) REIInh alation Toxicity In halation Exp osu re. -Fu rath iocarb Derm al Exposure stu dy DAT1 : u g/ hr DAT2 : u g/ hr Calculation of reentry in terval DFR0 = 35.3 ng/ cm 2 K = ng/ cm 2/ d ay TF = cm 2/ h r Available d ata ADDSL = 3500 ng/ kg/ d ay (=ADI) BW = 60 kg ABS = 20 % So. Reentry Interval T=[ln{( )/( )}-ln(35.3)] ( )-1 = 0.7 day(18 hours) Fu rathiocarb REI d ata Carbofu ran. Carbofu ran REI
28 18. Methid athion (g, ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 patch ug/ cm g/ h patch ug/ cm g/ h 3705 patch ug/ cm g/ h 33.3 patch ug/cm2 g/h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h ug/ cm g/ h ug/ cm g/ h
29 19. Methid athion (g, 1 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/cm2 g/h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h ug/ cm g/ h ug/ cm g/ h
30 20. Methid athion (g, 2 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 patch ug/ cm g/ h patch ug/ cm g/ h patch ND ND ug/ cm2 ND g/ h ND patch ug/ cm g/ h patch ND ND ug/ cm ND g/ h ND 24.8 patch ND ND 27.7 ug/ cm ND g/ h ND 6.7 patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ND ND ug/ cm2 ND g/ h ND ug/ cm g/ h ug/ cm g/ h
31 21. Methid athion (g, 3 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 patch ND ND ug/ cm ND g/ h ND 27.6 patch ND ND 5.4 ND ND ND 70.5 ND ug/ cm2 ND ND ND g/ h ND ND 19.3 ND patch ND ND ND ND ug/ cm2 ND ND g/ h ND ND ND ND ND ND patch ND 5.4 ND ND ND ND ND ug/ cm2 ND ND ND g/ h ND ND ND ND ND ND ND patch ug/ cm g/ h patch ug/ cm g/ h patch ND ND ug/ cm2 ND g/ h ND patch ND ND ug/ cm2 ND g/ h ND ND ND ug/ cm2 ND ND g/ h ND 33.7 ND ug/ cm g/ h
32 22. Methid athion (g, 5 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ug/ cm g/ h ND ug/ cm ND g/ h ND
33 23. Methid athion (g, 7 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND patch ND ND ND 0.4 ND ND ND 27.7 ug/ cm2 ND ND ND g/ h ND ND ND 6.9 ND ND ND ND patch ND ND 1.9 ND ND ND ND ug/ cm2 ND ND ND g/ h ND ND 26.9 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ug/ cm2 ND ND g/ h ND ND ND ug/ cm2 ND g/ h ND
34 24. Fu rathiocarb (g) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) patch ug/ cm g/ h patch ug/ cm g/ h 1883 patch ND ND ug/ cm2 ND g/h ND patch ug/ cm g/ h 19.1 patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h patch ug/ cm g/ h ug/ cm g/ h ug/ cm g/ h
35 25. Fu rathiocarb (g, 1 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 patch ND ND ND ND ND ND ug/cm2 ND ND ND g/h ND ND ND ND ND ND ND patch ND ND 4.6 ND ND ND 10.2 ND ug/ cm2 ND ND ND g/ h ND ND 2.5 ND ND ND ND ND ND ND ND ND ND ND ND ND patch ND 26.5 ND ND ND ND ND ug/ cm2 ND ND ND g/ h ND ND ND ND ND ND ND patch ND ND ND ND ug/ cm2 ND ND g/ h ND ND patch ND ND 26.5 ND ND ND ND ug/ cm2 ND ND ND g/ h ND ND 84.1 ND ND ND ug/ cm2 ND ND g/ h ND ND ug/ cm g/ h
36 26. Fu rathiocarb (g, 2 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 patch ND ND ug/ cm ND g/ h ND patch 3 ND 29.2 ND 39 ND ND ug/ cm ND ND g/ h 9.1 ND 87.7 ND patch ND 3.6 ND 4.2 ND 8.0 ND 9.2 ug/ cm2 ND ND g/ h ND 1.8 ND 2.1 patch ND ND ug/ cm2 ND g/ h ND ND ND ND ND ND ND ND ND patch ND 19.5 ND 7.9 ND ND ug/ cm2 ND 0.39 ND g/ h ND ND patch ND 6.4 ND ND ND ND ND ug/ cm2 ND ND ND g/ h ND 35.4 ND ND ND ND ND ND patch ND 8.9 ND 8.0 ND ND ug/ cm2 ND ND 0.04 g/ h ND 26.5 ND 23.7 ND 57.3 ND 39.3 ug/ cm2 ND ND 0.03 g/ h ND 13.2 ND ND ND ug/ cm ND ND g/ h ND ND
37 27. Fu rathiocarb (g, 3 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ug/ cm g/ h
38 28. Fu rathiocarb (g, 5 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ug/ cm g/ h
39 29. Fu rathiocarb (g, 7 ) (cm2) (650) (650) (150) (110) (3,550) (3,550) (2,910) (1,210) (3,820) (2,380) () (1,310) () (820) #1 #2 #3 #4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ug/ cm2 ND ND g/ h ND ND
40 30. m ethid athion furathiocarb (surfactant ) Methidathion Furathiocarb u g/ 405cm u g/ cm 2 Mean u g/ 405cm u g/ cm 2 Mean S.D. S.D. N D N D N D N D Con trol 1 Day 2 Day 3 Day 5 Day 7 Day N D N D N D N D N D N D N D N D N D ND ND N D
41 31. Methid athion furathiocarb (acetone ) Control 1 Day 2 Day 3 Day 5 Day 7 Day M ethid ath ion u g/ 405cm u g/ cm 2 N D N D Fu rathiocarb Mean u g/ 405cm u g/ cm 2 S.D. N D N D N D N D N D N D N D N D N D N D N D Mean S.D. N D
42 32. m ethid athion furathiocarb Control 1 Day 2 Day 3 Day 5 Day 7 Day Methidathion u g/ 50g (p p m ) ND ND F urathiocarb Mean u g/ 50g S.D. u g/ m l ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND Mean S.D. ND ND ND ND ND ND
43 2) Biological monitoring (1),, A. a) Methid athion b) Furathiocarb B. Methid athion Furathiocarb C6H 11N 2O4PS3 C18H 26N 2O5S colou rless crysta yellow viscous liqu i > 250 KOW, logp (25 ) 0.25mPa(20 ) 3.9 * 10-3m Pa(25 ) - Acetone - Water - Ethanol - H exane - Toluene 670g/ L(20 ) 200mg/ L(25 ) 150g/ L(20 ) 11g/ L(20 ) 720g/ L(20 ) read ily m iscible 11m g/ L(25 ) , DT50(water, 25 ) ( - 30 (ph 13) - - (< 400 ) 4 (ph 9)
44 C. Methid athion Fu rathiocarb N ovartis N ovartis (EC), (WP),, (GR,, cholinesterase inhibitor cholinesterase inhibito non-system ic insecticide, acaricid system ic insecticide D. Methid athion Furathiocarb ADI 0.001mg/ kg b.w - N OEL - rat - d og - hum an 0.15m g/ kg, d ail 0.25m g/ kg, d ail 0.11m g/ kg, d ail 0.35m g/ kg b.w WH O(a.i.) - EPA(formulation) Ib I Ib II (LD50) - rat, oral - rat, percutaneous - mou se, oral - rabbit, oral - rabbit, percutaneous - gu inea pig, oral - ham ster, oral - chicken, oral 25-54m g/ kg 1546m g/ kg 25-70m g/ kg 63-80m g/ kg 200mg/ kg 25mg/ kg 30mg/ kg 80mg/ kg 53m g/ kg > 2,000mg/ kg 327mg/ kg (LC50, rat, 4hr.) 3.6m g/ L air 0.214mg/ L air (LC50) - rainbow trou t(96hr.) - bluegill su nfish(96hr.) - lobster, nothern(48hr.) - d aphnia(48hr.) - rainbow trou t, carp, blu egill sunfish( mg/ L 0.002m g/ L 0.014m g/ L ug/ L m g/
45 (2), A. Methid athion : technical(96.2%), emu lsifiable concentrate(40% EC) Furathiocarb : technical(97.8%), emu lsifiable concentrate(10% EC), w ettable pow d er(10% WP) H yd rochloric acid, Sod iu m chlorid e, Sod ium su lfate : GR, Junsei(Jap an) H exane, Acetone, Ethyl acetate, Dichlorom ethane : GR, Junsei(Japan) Water, Methanol : H PLC grad e, Du ksan(korea) Silica gel : colum n chrom atography grad e, Merck(Germany) Ep pend orf tube : Sarsted t(usa) Mem brane filter : Acrodisc LC13 PVDF filter(13mm, 0.45um, Gelman,USA) FP Vericel filter(47m m, 0.45u m, Gelm an, USA) B. 7-8 S.D. rat( )4 ( g). 23 2, 50 10%, 12(7-7 ), cage 3-4. C. Animal clip per : Oster Pro-power E(Oster, USA) Glass d iffusion cell : Franz cell Tissue homogenizer : Ultra Tu rrax T25 basic(ika Lab., Germ any) Balance : H M-200 & H F-300GD(AND, Jap an) Vortex mixer : KMC-1300V(Vision, Korea) Centrifuge : Sup ra 22K(H anil, Korea), VS 5000(Vision, Korea) Sep aratory funnel shaker : ST-0(Woo Ju, Korea) N itrogen evaporator : Reacti-Vap & Reacti-Therm (Pierce, USA) Rotary vacuu m evap orator : Rotavap or R-114(Bu chi, Sw itzerland ) H PLC : H ew lett Packard series 1100 H PLC(H P, Germ any) GC : H ew lett Packard 5890 series II p lu s GC(HP, USA)
46 (3) SD rat ether,.,.,.. (a-cyanoacrylate), (, 2cm )., (0.9% saline solu tion) 18m L glass bead incubation, , m ethid athion 100mg acetone 20, 50, 100, 200, 500uL. (250rp m) (2, 4, 6, 8, 12, 18, 24 ) 500u L 500u L. mem brane filter, H PLC autosamp ler 10uL chromatogram p eak m ethid athion, , acetone 200uL m ethid athion 20, 50, 100, 200mg. (250rpm ) (2, 4, 6, 8, 12, 18,
47 ) 500u L 500u L. m embrane filter, H PLC au tosamp ler 10uL chromatogram p eak methid athion, Methid athion methid athion, m ethid athion 100m g acetone 200u L. (250rp m) (2, 4, 6, 8, 12, 18, 24, 36, 48 ) 500uL 500uL. mem brane filter, H PLC au tosam pler 10uL chromatogram p eak m ethid athion, Methid athion methid athion (40%), m ethid athion 250u L(methidathion, 100m g) (250rp m) (2, 4, 6, 8, 12, 18, 24, 36, 48 ) 500u L 500uL. m embrane filter, H PLC au tosamp ler 10u L, , 50, 100, 200, 500uLmethid athion (Figure 5), 20u L100uL
48 m ethid athion, 100u L. 20u L 50uL., 100uL,, 200uL , 50, 100, 200mg methid athion (Figure 6), 20m g, 50mg. 50mg m ethid athion. 100mg
49 Methid athion Methid athion (3.14cm 2) ( 33). Methid athion (Figure 7) (Figure 8) 13.92u g/ cm 2/ hr RSD = 2.49) 15.28ug/ cm2/ hr(rsd = 4.60), m ethid athion., 24, methid athion (200m g/ L w ater)., 24, Methid athion (: pp m)
50 - 43 -
51 ) Methid athion Instrument : H P 1100 series H PLC(H P, Germ any) Colu mn : Zorbax SB-C18 colu mn(250 x 4.6mm, 5u m, 30, H P) Detection : UV230nm Mobile phase : 70% m ethanol in w ater Flow rate : 1.0m l/ min Injection volu m e : 10u L (LOD, limit of detection) (LOQ, limit of quantification) 1) Methid athion (LOD) : 0.03pp m(0.3ng) (LOQ) : 0.1p pm (1ng) ) Methid athion Methid athion m ethanol 1, 2, 5, 10, 20, 50, 100p pm, 10u L H PLC chrom atogram p eak (Figure 9)
52 (4) Biological m onitoring I 4-1. Methid athion x 6cm 2.5 x 2.5cm LD50 3/ 4, 3/ 20, 3/ mg/ kg, 231.9m g/ kg, mg/ kg. metabolic cage2, 4, 6, 8, 12, 24 48,,, Methid athion 1) 500u L 1.5mL Ep pend orf tu be 6N H Cl 100u L. hexane 500u L30 2 vortexing, 4, 20,000 rpm 10. GC autosamp ler hexane 5u L chromatogram peak m ethid athion. 2) 500u L 1.5mL Epp end orf tube hexane 500u L30 2 vortexing, 4, 15,000 rp m10. GC autosam pler hexane 5uL chrom atogram p eak methid athion, m ethid athion. 3) hom ogenizer. 400m g 1.5m L Ep pend orf tu be400ul vortex mixer
53 . hexane 400u L30 3 vortexing, 4, 20,000 rpm 10. H exane 250u L, hexane 50u L. GC au tosam pler hexane 5uL chrom atogram p eak methid athion(5 ). 4), homogenizer. 400mg 1.5m L Ep pend orf tu be. hexane 400u L30 3 vortexing, 4, 20,000 rp m10. GC autosam pler hexane 5uL chrom atogram p eak methid athion (2 )
54 m ethid athion m ethid athion LD50 3/ 4, 3/ 20, 3/ 40, 12, 8, 6 ( 34, Figure 10). 34. Methidathion (: ppm) LD50, / 4 RSD LD50, 3/ 20 LD50, 3/ RSD RSD
55 methid athion methid athion m ethid athion ( 35, Figu re 11). 35. Methid athion (: u g) LD50, / 4 RSD LD50, 3/ 20 LD50, 3/ RSD RSD
56 methid athion methid athion 8 ( 36, Figu re 12) 36. Methidathion,p pm LD50, (RSD) 3/ 4,ug (RSD),p pm LD50, (RSD) 3/ 20,ug (RSD),p pm LD50, (RSD) 3/ 40,ug (RSD) (16.64) (18.68) (9.03) (21.07) (14.14) (14.14) (25.25) (7.95) (24.27) (28.28) (13.54) (10.10)
57 methid athion methid athion 8 ( 37, Figure 13) 37. Methidathion,p pm LD50, (RSD) 3/ 4,ug (RSD),p pm LD50, (RSD) 3/ 20,ug (RSD),p pm LD50, (RSD) 3/ 40,ug (RSD) (17.49) (21.04) (8.37) (2.76) (21.76) (5.79) (13.51) (3.81) (5.06) (17.10) (5.44) (6.96) (27.78) (16.50)
58 Instrument : H P 5890 series II GC(HP, USA) Colu mn : H P-5 capillary colu mn(30 x 0.53mm, 1.5u m, H P) Temp eratu re : Injector(240 ), Colu m n oven(210 ), Detector(260 ) Flow rate : 20mL/ m in(carrier gas, nitrogen) Injector : Packed inlet Detector : FPD(P filter) Injection volu m e : 5u L Retentiom tim e : 4.6min (LOD, limit of detection) (LOQ, limit of quantification) (LOD) (LOQ) 0.01p pm 0.01pp m 0.002p pm 0.02p pm (0.05ng) (0.05n g) (0.01n g) (0.1ng) 0.03p pm 0.03pp m 0.006p pm 0.06p pm (0.15ng) (0.15n g) (0.03n g) (0.3ng) Methid athion hexane 0.02, 0.05, 0.1, 0.2, 1, 2pp m, 5uL GC chrom atogram p eak (Figure 14)
59 ),, 500 ul methid athion 0.05, 0.5, 2pp m,. m ethid athion % % ( 38). 38. m ethid athion (pp m) (%) RSD(, %) ),, 400m g methid athion 0.02, 0.05, 0.1pp m, 0.05, 0.1, 0.5p pm,. methid athion % % ( 39). 39. m ethid athion (p pm ) (%) RSD(, %)
60 4-2. Furathiocarb x 6cm 2.5 x 2.5cm m g/ kg, 300.0mg/ kg, 150.0mg/ kg. metabolic cage2, 4, 6, 8, 12, 24 48,,, Furathiocarb 1) 700uL 1.5mL Ep pend orf tu be, (EtOAc:H exane=75:25) 700u L30 2 vortexing, 4, 20,000 rpm u L N 2, methanol 125uL 0.45u m m embrane filter H PLC autosamp ler 10u L chromatogram p eak furathiocarb (4 ). 2) 700u L 1.5m L Epp end orf tu be, (EtOAc:H exane=75:25) 700u L30 2 vortexing, 4, 20,000 rpm u L N 2, methanol 125uL 0.45u m m embrane filter H PLC autosamp ler 10u L chromatogram peak furathiocarb, furathiocarb
61 3) hom ogenizer. 500m g 20m L vial acetone 5m L vortex mixer, 30. sod ium sulfate, acetone 5m L vial. Acetone, 5m L. ethyl acetate 5m L30 2 vortexing, 4,000 rpm 10. Ethyl acetate 2.5mL, 10% EtOAc(in hexane) 1m L. silica gel 10gcolumn, 5% EtOAc(in hexane) 50mL 10% EtOAC(in hexane) 70mL. 50% EtOAc(in hexane) 150mL, methanol 250u L, H PLC autosamp ler 10uL chromatogram p eak furathiocarb. 4), homogenizer. 1g 20mL vial acetone 5m L vortex m ixer, 30. sod iu m su lfate, acetone 5mL vial. Acetone, 5m L. ethyl acetate 5m L30 2 vortexing, 4,000 rpm 10. Ethyl acetate 2.5m L, 10% EtOAc(in hexane) 1mL. silica gel 10gcolum n, 5% EtOAc(in hexane) 50m L 10% EtOAC(in hexane) 70m L. 50% EtOAc(in hexane) 150mL, m ethanol 250u L, H PLC autosam pler 10u L chrom atogram peak furathiocarb
62 furathiocarb furathiocarb 1,500m g/ kg carbofuran(cf) 3-hydroxycarbofuran(3-HC), 300m g/ kg carbofu ran. 150mg/ kg fu rathiocarb ( 40, Figure 15, 16). 15. Furathiocarb (: ppm) CF 300 mg/ kg Total FC* CF 1,500 3-HC mg/ kg Total FC* RSD RSD RSD RSD RSD *) Total FC : CF 3-H C FC
63 - 56 -
64 furathiocarb furathiocarb carbofuran 3-hydroxycarbofuran, furathiocarb ( 40, Figu re 17, 18, 19). 40. Fu rathiocarb (: u g) CF HC mg/ kg Total FC* CF HC mg/ kg Total FC* CF 1,500 3-HC mg/ kg Total FC* RSD RSD RSD RSD RSD RSD RSD RSD RSD *) Total FC : CF 3-H C FC
65 - 58 -
66 - 59 -
67 fu rathiocarb fu rathiocarb 8 ( 41, Figu re 20) 41. Fu rathiocarb CF 1,500 3-HC mg/ kg Total FC* RSD RSD RSD *) Total FC : CF 3-H C FC
68 furathiocarb furathiocarb 8 ( 42, Figure 21) 42. Fu rathiocarb CF 1,500 3-HC mg/ kg Total FC* RSD RSD RSD *) Total FC : CF 3-H C FC
69 Instrument : H P 1100 series H PLC(H P, USA) PCX 5100 post-column derivatization instrument(pickering Lab., USA) Colu mn : Carbamate analysis column(c18, 150 x 4.6mm, 5um, Pickering Lab.) Mobile phase : 20% MeOH (1min) --> 55% MeOH (20m in) --> 90% MeOH (22-28m in) --> 100% MeOH (30-40m in) Flow rate : 1.0m l/ min PCX 5100 p ost-column d erivatization instru m ent - Reagent A : H yd rolysis reagent - Reagent B : OPA reagent - Flow rate : 0.3mL/ m in - Temp eratu re : H yd rolysis(100 ), Colu mn(42 ) Detector : Flu orescence d etertor(ex. 330nm, Em. 465nm) - Gain : 10 - Response : 2sec - Frequency : 220H z Injection volu m e : 10u L Retentiom tim e : 3-Hydroxycarbofuran(3-HC, 11.7min), Carbofuran(CF, 19.8min) 3-Ketocarbofu ran(3-kc, 15.9m in), Fu rath iocarb(26.7m in) (LOD, limit of detection) (LOQ, limit of quantification) 3-HC 0.025pp m(0.25ng 0.025p pm (0.25ng 0.1p pm (1ng) 0.1pp m(1ng) 3-KC 0.05pp m(0.5ng) 0.05p pm (0.5ng) 0.2p pm (2ng) 0.2pp m(2ng) CF 0.025pp m(0.25ng 0.025p pm (0.25ng 0.1p pm (1ng) 0.1pp m(1ng) (LOD) FC 0.05pp m(0.5ng) 0.05p pm (0.5ng) 0.2p pm (2ng) 0.2pp m(2ng) 3-HC 0.075pp m(0.75ng 0.075p pm (0.75ng 0.3p pm (3ng) 0.3pp m(3ng) 3-KC 0.15pp m(1.5ng) 0.15p pm (1.5ng) 0.6p pm (6ng) 0.6pp m(6ng) CF 0.075pp m(0.75ng 0.075p pm (0.75ng 0.3p pm (3ng) 0.3pp m(3ng) (LOQ) FC 0.15pp m(1.5ng) 0.15p pm (1.5ng) 0.6p pm (6ng) 0.6pp m(6ng)
70 Fu rathiocarb, carbofu ran, 3-hyd roxycarbofu ran, 3-ketocarbofu ran m ethanol 0.2, 0.5, 1, 2pp m, 10u L H PLC chrom atogram peak (Figure 22) ),, 700 ul furathiocarb 0.5, 1p pm,. 2), 500m g 1g furathiocarb 0.5pp m 1p pm,
71 43. fu rathiocarb (%) RSD( (ppm) 1 2 3, %) 3-HC 0.125p p m p p m KC 0.125p p m p p m CF 0.125p p m p p m FC 0.125p p m p p m HC 0.125p p m p p m KC 0.125p p m p p m CF 0.125p p m p p m FC 0.125p p m p p m furathiocarb (%) RSD( (pp m) 1 2 3, %) 3-H C 0.5p p m p p m KC 0.5p p m p p m CF 0.5p p m p p m FC 0.5p p m p p m H C 0.5p p m p p m KC 0.5p p m p p m CF 0.5p p m p p m FC 0.5p p m p p m
72 (5) : :,. PBPK Methid athion Methid athion 22m g 1mL DMSO 100mL. 220pp m m ethid athion 1% DMSO H PLC, m ethid athion (,, ) g3ml hom ogenizer mL220pp m methid athion 100u L0 1, 2, 4, 6, 8, 10. Methid athion 30,000g, u m mem brane filter m ethid athion H PLC methid athion
73 ,, 1mL220p pm m ethid athion 100uL0 5. methid athion 20pp mdmso 0.1%. Methid athion 30,000g, u m mem brane filter m ethid athion H PLC methid athion. 1mL m ethid athion 100u L 5, 0.45u m m embrane filter m ethid athion H PLC, m ethid athion. (Ptw, tissu e:w ater partition coefficient)., Ptw = ( methidathion, Ct)/ ( methidathion, Cw), m ethid athion. m ethid athion = {( methidathion ) -( methidathion )*( )} / (), (Pbw ; blood :w ater partition coefficient). Ptw Pbw : (P) P = Ptw / Pbw Methid athion p pm methid athion, 0-24 methid athion. m ethid athion
74 Figure 23 m ethid athion 4. :, :
75 Methid athon : 8. 9: methidathion : (SD) Tissu e Ct Cw Ptw = Ct/ Cw (p pm RSD (pp m RSD RSD Liver Kid ney Blood Tissue:blood p artition coefficients Tissu e P = Ptw / Pbw Liver, PL 2.82 Kid n ey, PK
76 (6) Biological m onitoring II - Field test 6-1. Methid athion Methid athion 1) 500uL 1.5mL Ep pend orf tu be 6N H Cl 100u L. hexane 500u L30 2 vortexing, 4, 20,000 rpm 10. H exane 250u L, hexane 50u L. GC au tosam pler hexane 5uL chromatogram p eak methid athion(5 ). 2) 50m L 250mL 50m L N acl 20g. Dichloromethane 50mL mL, sod iu m su lfate 4 0. H exane 1mL GC au tosam pler hexane 5uL chromatogram p eak methid athion(50 )
77 methid athion (: p pm ) N ame Date Comm ent Average(n=3) RSD 5/ / / / 12 Blan k / / / / / / / / / / / methid athion N am e Date Time Com ment urine(m L) concentration qu antity (ppm ) (ug) RSD 5/ 13 13:30 1, / 14 7:30 2, :00 2, / 15 8:15 3, / 13 19:00 1, :00 2, / 14 12:44 2, :00 2, / 15 7:30 3, / 13 19:00 1, / 14 6:50 2, :50 2, / 15 7:30 3, / 13 19:00 1, / 14 5:30 2, :00 2, / 15 6:00 3,
78 (LOD, limit of detection) (LOQ, limit of quantification) (LOD) (LOQ) 0.002p pm (0.01ng p pm (0.001ng 0.006p pm (0.03ng p pm (0.003ng 6-2. Furathiocarb Furathiocarb 1) 700uL 1.5m L Ep pend orf tu be, (EtOAc:H exane=75:25) 700uL30 2 vortexing, 4, 20,000 rpm u L N 2, methanol 125uL 0.45u m m embrane filter H PLC autosamp ler 10u L chromatogram p eak furathiocarb (4 ). 2) 10m L 125m L 50m L. Dichlorom ethane 50m L mL, sod iu m su lfate 40. Methanol 200uL H PLC au tosamp ler 10u L chrom atogram peak fu rathiocarb (50 )
79 furathiocarb (: p pm ), fu rathiocarb. N ame Date Com ment Concentration(ppm < / 2 < < / 3 < / 4 < < / 3 < < / 4 < / 3 < < / 4 < / 3 < < / 4 <
80 fu rathiocarb., carbofuran N am e Date Tim e Com m ent Carbofuran Urin e(m L) Concentration Qu antity (ppm ) (ug) 6/ 2 20:00, 120 < / 3 7:00 1, 340 < / 3 16:00 1, / 3 21:00 1, / 4 7:00 2, 390 < / 4 16:30 2, 240 < / 4 22:00 2, 260 < / 5 7:00 3, 330 < / 3 21:30 1, / 4 7:00 2, / 5 7:30 3, 220 < / 3 19:40 1, 280 < / 4 7:00 2, / 4 7:10 2, 200 < / 5 6:00 3, 460 < / 4 5:45 2, / 4 19:43 2, 210 < / 5 6:25 3, 400 < (LOD, lim it of d etection) (LOQ, limit of qu antification) 3-H C 0.025pp m(0.25ng 0.002pp m(0.02ng 3-KC 0.05p p m(0.5ng) 0.004p p m(0.04n g CF 0.025pp m(0.25ng 0.002pp m(0.02ng (LOD) FC 0.05pp m(0.5ng) 0.004pp m(0.04ng 3-H C 0.075pp m(0.75ng 0.006pp m(0.06ng 3-KC 0.15p p m(1.5ng) 0.012p p m(0.12n g CF 0.075pp m(0.75ng 0.006pp m(0.06ng (LOQ) FC 0.15pp m(1.5ng) 0.012pp m(0.12ng
81 (7) Kw ang-hyeon Liu, Ji-Youn Byou n, H a-ju ng Sung, H ae-keun Lee and Jeong-H an Kim., Derm al Penetration Rate of Meth id athion - In vitro -, 1999, Kw ang-h yeon Liu, Jeong-H an Kim, H a-jung Su ng and H ae-keun Lee Dermal Penetration of Methidathion, 1999,
82 : Biological monitoring : : 96 99(3 ), 3, - ( 97 ) - ( 97 ) - biological m onitoring ( 98 ) - ( 98 ) - ()
83 - 1: - 2: - 3: (, ),, - - < > biological m onitoring. - : SPF S.D. - : Methid athion, furathiocarb , - ChE
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