. 1999. 11.. : : : :, :,,,,, :,, : : :, :
------------------------------------------------------------------------------------ 1 ------------------------------------------------------------------------------------ 5 1. ---------------------------------------------------------------------------------------- 5 1) --------------------------------------------------------------- 5 2) Biological m onitoring ---------------------------------------------------------- 5 3) --------------------------------------------------------- 6 4) mod el ------------------------- 6 2. ---------------------------------------------------------------------------- 7 1) ----------------------------------------------------------------- 7. (Method valid ation) ------------------------------------------------ 7 (1) (LOD) (LOQ) --------------------------------------- 7 (2) (Reproducibility test) -------------------------------------- 7 (3) (Stand ard calibration cu rve lin erity) ------------------------------------------- 7 (4) (Storage stability test) --------------------------------------- 8 (5) (Breakthrou gh test) --------------------------------------------- 9 (6) (Trapping efficiency test) --------------------------------- 9. (Recovery test) ---------------------------------------------------------- 10 (1) ---------------------------------------------------------------------------- 10 (2) (Field recovery test) ------------------------------------------ 11 (3) -------------------------------------------------------------------- 12 (4) surfactant(sur-ten ) ----------------------- 13 (5) (acetone) -------------------------- 14. ----------------------------------------------------------------------------------- 15
(1) ----------------------------------------------------------------------- 15 (2) ----------------------------------------------------------------------- 16 (3) ------------------------------------------------------------------- 16 (4) ---------------------------------------------------- 16 (5) ---------------------------------------------------------------------- 19 2) Biological m onitoring ---------------------------------------------------------- 36 (1),, -------------------- 36 (2), ----------------------------------------------------- 38 (3) ----------------------------------------------------------------------- 39 3-1. ------------------------------------------------------------------------------- 39 3-1-1. --------------------------------------- 39 3-1-2. ------------------------------------------- 39 3-1-3. ------------------------------------------------ 39 3-1-4. Methid athion ---------------------------------------- 40 3-1-5. Methid athion ---------------------------------------- 40 3-2. --------------------------------------------------------------------------------- 40 3-2-1. --------------------------------------------- 40 3-2-2. ------------------------------------------ 41 3-2-3. Methid athion ------------------------------ 42 3-2-5. --------------------------------------------------------------------- 44 3-2-6. -------------------------------------------------------------- 44 3-2-7. ------------------------------------------------------------------ 44 (4) Biological m on itorin g I ----------------------------------------------------------------- 45 4-1. Methid ath ion ---------------------------------------------------------------------------- 45
4-1-1. ----------------------------------------------------------------------------- 45 4-1-1-1. ---------------------------------------- 45 4-1-1-2. Methid athion --------------------------------------------- 45 4-1-2. ------------------------------------------------------------------------------ 47 4-1-2-1. methid athion --------------------------------------- 47 4-1-2-2. methid athion----------------------------------------------- 48 4-1-2-3. methid athion ------------------------------------------- 49 4-1-2-4. methid athion --------------------------------------- 50 4-1-3. ----------------------------------------------------------------------- 51 4-1-4. (LOD) (LOQ) ---------------------------------------------- 51 4-1-5. -------------------------------------------------------------------- 51 4-1-6. --------------------------------------------------------------------------- 52 4-2. Fu rathiocarb ------------------------------------------------------------------------------- 53 4-2-1. -------------------------------------------------------------------------------- 53 4-2-1-1. ----------------------------------------- 53 4-2-1-2. Fu rathiocarb --------------------------------- 53 4-2-2. -------------------------------------------------------------------------------- 55 4-2-2-1. furathiocarb ---------------------------- 55 4-2-2-2. furathiocarb ----------------------------------- 57 4-2-2-3. furathiocarb ------------------------------ 60 4-2-2-4. furathiocarb ---------------------------- 61 4-2-3. ----------------------------------------------------------------------- 62 4-2-4. --------------------------------------------------------------- 62 4-2-5. -------------------------------------------------------------------- 63 4-2-6. --------------------------------------------------------------------------- 63
(5) : ----------------------------------------------------------------- 65 5-1. ---------------------------------------------------------------------------------- 65 5-2. ---------------------------------------------------------------------------------- 65 5-2-1. Methid ath ion ------------------------------------------------------------------------ 65 5-2-1-1. ----------------------------------------------------------------- 65 5-2-1-2. ----------------------------------------------------------------------- 65 5-2-1-3. ----------------------------------------------- 65 5-2-1-4. ----------------------------------------------- 66 5-3. ---------------------------------------------------------------------------------- 66 5-3-1. Methid ath ion ------------------------------------------------------------------------- 66 5-3-1-1. -------------------------------------------------------------- 66 5-3-1-2. ------------------------------------------------------- 67 5-3-1-3. ------------------------------------------------------ 68 (6) Biological m on itorin g II - Field test ------------------------------------------------ 69 6-1. Meth id athion ----------------------------------------------------------------------------- 69 6-1-1. ------------------------------------------------------------------------------ 69 6-1-1-1. -------------------------------------------------------------------------- 69 6-1-1-2. -------------------------------------------------------------------------- 69 6-1-1-3. Methid athion ----------------------------------------------- 69 6-1-2. -------------------------------------------------------------------------------- 70 6-1-2-1. methid athion ----------------------------------------- 70 6-1-2-2. methid athion ------------------------------ 70 6-1-2-3. ----------------------------------------------------------- 71 6-2. Fu rathiocarb ------------------------------------------------------------------------------- 71 6-2-1. -------------------------------------------------------------------------------- 71
6-2-1-1. ---------------------------------------------------------------------------- 71 6-2-1-2. ---------------------------------------------------------------------------- 71 6-2-1-3. Fu rathiocarb ------------------------------------------------ 71 6-2-2. -------------------------------------------------------------------------------- 72 6-2-2-1. furathiocarb --------------------------- 72 6-2-2-2. fruathiocarb ----------------- 73 6-2-2-3. ----------------------------------------------------------- 73 (7) -------------------------------------------------------------------------------------- 74 : Biological m onitoring ----------------------------------- 75 ------------------------------------------------------------------------- 75 ------------------------------------------------------------------------------------------------ 75 --------------------------------------------------------------------------- 75 --------------------------------------------------------------------------------- 75 ------------------------------------------------------------------------------------------------ 75 ------------------------------------------------------------------------------------------- 76 ------------------------------------------------------------------------------------------------ 76 Methid athion ChE ---------------------------------------- 77 Furathiocarb ChE ----------------------------------------- 80 ------------------------------------------------------------------------------------------- 83 --------------------------------------------------------------------------------------- 83 ------------------------------------------------------------------------------------- 83 1. ----------------------------------------------------------- 84
2 82-92 13 57%, 77%..,,. (REI; Reentry interval)..,,,.. (89 93 ) 3,202,0004,756. 40 94%, 1/ 10. 95 108 605, 1994 26,282 M/ T. - 1 -
,,.,. (EPA).. 94 80,453ha.,, 96 (, )37,848 ha 13.0%(4,352 ha).,,,,, 90 23,698 ha 34 ha. ().,,,,,,. 6 4 1 28. 4 5 8( 15 ) - 2 -
.,,..,,,.. 2,,..,.,,,,, biological m onitoring, m anu al, mod el., p atch,, DDVP, bifenthrin., PBPK mod el EPA. - 3 -
. - 4 -
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,. - 5 -
,. 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). - 6 -
1). (Method validation) (1) (LOD ) (LOQ) (LOD) (LOQ) ( 1). 1. Pes ticide LOD(ng ) LOQ(ng) Methidathion 0.05 0.25 F urathiocarb 0.05 0.25 (2) (Reproducibility test) CV 0.7 7.2%, ( 2). 2. pesticide Methidathion Furathiocarb Concentra tion(ppm) Height 1 2 3 4 5 6 Averag e CV(% ) SD 0.1 0.726 0.721 0.712 0.706 0.680 0.694 0.707 0.017 2.4 1.0 8.642 8.680 8.802 8.723 8.797 8.704 8.725 0.064 0.7 10 71.838 71.564 70.505 71.330 73.378 71.010 71.604 0.984 1.4 0.1 0.215 0.258 0.235 0.228 0.212 0.231 0.230 0.017 7.2 1.0 2.630 2.621 2.672 2.660 2.621 2.629 2.639 0.022 0.8 10 23.525 23.414 23.185 22.647 23.077 22.947 23.133 0.319 1.4 (3) (Standard calibration curve linearity) lin earity 4 ( 3). - 7 -
3. Pesticide Calibration curv e 1 3 Methidathion y=3.2515x- 0.0624, r=0.9994 y=2.3719x- 0.1242, r=0.9999 y=1.9698x+0.3086, r=0.9989 Furathiocarb y=2.4307x- 0.1038, r=0.9979 y=6.7227x- 0.3274, r=0.9992 y=5.7197x- 0.4254, r=0.9981 (4) (Storage stability test). 7. ( 4, 5, 6, 7) 4. Patch Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%) 50 33.6 36.9 35.1 35.2 1.65 4.69 70.4 1 50 39.3 38.7 39.6 39.2 0.46 1.17 78.4 3 50 38.1 35.4 34.5 36.3 1.87 5.16 72.6 7 50 39.9 38.7 40.5 39.7 0.92 2.31 79.4 50 34.5 37.2 36.0 35.9 1.4 3.8 71.8 1 50 35.7 36.0 37.2 36.3 0.8 2.2 72.6 3 50 36.9 35.4 34.5 35.6 1.2 3.4 71.2 7 50 40.0 38.7 39.9 39.5 0.7 1.8 79.0 5. Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%) 500 408 420 438 422 15.1 3.6 84.4 1 500 354 387 372 371 16.5 4.5 74.2 3 500 327 387 375 363 31.7 8.7 72.6 7 500 495 360 498 451 78.8 17.5 90.2 500 381 402 399 394 11.4 2.9 78.8 1 500 426 426 447 433 12.1 2.8 86.6 3 500 414 429 426 423 7.9 1.9 84.6 7 500 429 480 531 480 51 10.6 96-8 -
6. Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%) 500 384 396 369 383 13.5 3.5 76.6 1 500 408 408 393 403 8.7 2.2 80.6 3 500 420 402 417 413 9.6 2.3 82.6 7 500 417 450 471 446 27.2 6.1 89.2 500 462.6 435 452.1 449.9 13.9 3.1 90.0 1 500 422.7 423.6 432.3 426.2 5.3 1.2 85.2 3 500 423 408 417 416 7.5 1.8 83.2 7 500 384 384 387 385 1.7 0.4 77.0 7. XAD Pesticide Methidathion Furathiocarb (ug) (ug ) % CV Recovery (%.) 100 73.6 74.8 74.8 74.4 0.69 0.93 74.4 1 100 80 80 81 80.3 0.58 0.72 80.3 3 100 77.8 81.8 82 80.5 2.37 2.94 80.5 7 100 80.2 87.4 83.2 83.6 3.62 4.33 83.6 100 80 82.2 80.6 80.9 1.1 1.4 80.9 1 100 72.2 74.2 74.6 73.7 1.3 1.7 73.7 3 100 76.8 71.8 70 72.9 3.5 4.8 72.9 7 100 78.6 78 76.2 77.6 1.2 1.6 77.6 (5) (Breakthrough test) 2. ( 8) 8. (ug) Methidathion 152 F urathiocarb 50 (ug ) CV 1 2 (%) (%) T rap T rap 157.6 ND 103.6 171.1 ND 167.2 8.4 5.0 112.3 109.9 5.47 172.9 ND 113.7 51.4 ND 102.8 49.09 ND 49.03 2.4 0.04 98.18 98.06 4.8 46.6 ND 93.2 (6) (Trapping efficiency test) - 9 -
XAD. ( 9) 9. (ug) (ug) CV (% ) (%) (%) Methidathion 500 F urathiocarb 100 1 374.6 89.9 464.5 92.9 2 501.0 44.0 545.0 536.9 68.7 12.7 109 3 569.8 31.4 601.2 120.24 1 12.3 83.8 96.1 96.1 2 33.5 77.6 111.1 93.0 19.9 21.4 111.1 3 24.7 47 71.7 71.7 107.4 93.0. (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 1.5 1.3 1.5 1.7 1.5 0.2 0.11 99.3 10LOQ 15 13.4 14.5 13.7 13.9 0.57 4.1 92.4 100LOQ 150 141 142.5 142.7 142.1 0.93 0.7 94.7 LOQ 2.5 2.4 2.1 2.1 2.2 0.2 0.08 88 10LOQ 15 13.8 13.8 12.9 13.5 0.5 0.04 90 100LOQ 150 141.3 143.4 147.6 144.1 3.2 0.02 96.1 11. - 10 -
Methidathion Furathiocarb (ug ) (ug ) % CV Recovery (%) LOQ 7.5 6.7 7.3 6.9 7.0 0.3 4.4 92.9 10LOQ 75 78.6 84.3 85.5 82.8 3.7 0.04 110.4 100LOQ 750 732 786 750 756 27.5 0.04 100.8 LOQ 15 19.5 17.4 15.9 17.69 1.8 0.10 117.3 10LOQ 100 66.0 84.0 90.0 80.0 12.5 0.16 80.0 100LOQ 1000 957.0 1017 987.0 987.0 30 0.03 98.7 12. Methidathion Furathiocarb (ug ) (ug ) % CV Recovery (%) LOQ 15 17.1 15 17.4 16.5 1.31 0.08 110 10LOQ 150 160.5 144 154.8 153.1 8.38 0.05 102.1 100LOQ 1500 1353 1323 1296 1324 28.51 0.02 88.3 LOQ 15 14.1 15.7 13.8 14.5 1.0 7.1 93.8 10LOQ 150 126 123 129 126 3 0.02 84 100LOQ 1500 1194 1371 828 1131 276.9 0.2 75.4 13. XAD Methidathion Furathiocarb (ug ) (ug ) % CV Recovery (%) LOQ 5 3.8 3.7 4.4 4.0 0.4 9.5 76.5 10LOQ 50 45.2 47.1 41.8 44.7 2.7 6.0 90.5 100LOQ 500 416.0 446.1 444.1 435.4 16.9 3.9 83.2 LOQ 2.5 2.2 2.1 2.3 2.2 0.1 0.05 88 10LOQ 12.5 11.5 11.5 11.6 11.5 0.06 0.005 92.3 100LOQ 125 119.5 123.6 118 120.4 2.9 0.02 96.3 (2) (Field recovery test) - 11 -
. Meth id ath ion XAD ( 14). 14. (ug ) Methidathion Furathiocarb (ug) 1 2 3 (% ) Patch 1.5 1.3 1.7 1.5 1.5 0.2 99 75 78.6 84.3 85.5 82.8 3.7 110.4 150 160.5 144.0 154.8 153.1 8.38 102.1 XAD 100 80 81 80 80.3 0.58 80.3 Patch 15 13.8 13.8 12.9 13.5 0.5 90 100 66.0 84.0 90.0 80.0 12.5 80 150 126.0 123.0 129.0 126.0 3.0 84 XAD 12.5 11.5 11.5 11.6 11.53 0.06 92.3 (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) - 12 -
15. Spiked level(ug) Methidathion (ug ) (%) Mean S.D. Spiked lev el(ug ) Furathiocarb (ug ) (%) Control - ND - - - ND - - 1 12.5 2 125 3 2500 11.5 92 3.0 120 11.9 95 94.27 1.97 2.5 2.0 80 11.9 95 2.5 100 101.0 80 10.0 80 106.5 85 81.73 3.11 12.5 12.0 96 99.0 79 9.5 76 1855 74 90.0 72 2055 82 79.80 4.87 125 95.0 76 2075 83 95.0 76 Mean S.D. 100.0 20.0 84.0 10.58 74.67 2.31-85.27 3.35-86.22 10.96 (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 3. 0.02% 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).. - 13 -
16. su rfactan t(sur-ten ) Spiked level(ug ) Methidation (ug) (%) Mean S.D. (ug) Furathiocarb (ug) (%) Control - ND* - - - ND** - - LOQ 1.0 10 LOQ 10 100 LOQ 100 1.02 102 0.97 97 1.05 105 101 5.1 1.0 0.89 89 0.95 95 0.94 94 11.6 116 11.6 116 11.8 118 118 2.0 10 10.5 105 12.0 120 11.4 114 105 105 104 89 93 93 103 8.7 100 114 114 Mean S.D. 93 4.0 112 5.9 105 8.1 110 110 93 93-107 5.3 103 6.0 * 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 100 120 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). - 14 -
17. (aceton e) Spiked level(ug ) Methidation (ug) (%) Mean S.D. Spiked level(ug ) Furathiocarb (ug) (%) Control - ND* - - - ND** - - LOQ 1.0 10 LOQ 10 100 LOQ 100 0.75 75 0.71 71 0.84 84 79 4.6 1.0 0.75 75 0.78 78 0.78 78 8.3 83 7.4 74 9.4 94 89 5.7 10 8.0 80 9.1 91 8.3 83 82 82 77 77 92 92 84 7.2 100 83 83 78 78 88 88 Mean S.D. 75 3.5 79 4.6 82 5.5-84 5.8-78 4.5 * 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. - 15 -
1. Methidathion D ermal exposure 2. Furathiocarb D ermal exposure - 16 -
(2). (3),. m u lching. (4) 1methid athion 222.6u g/ cm 2, furathiocarb 35.3ug/ cm 2, 7 12.1u g/ cm2, 2.6u g/ cm2. (3, 4).. su rfactant acetone m ethid athion 1 14.2ug/ cm2, 2 12.4 u g/ cm2, 3 8.3u g/ cm 2, 5 2.1ug/ cm 2, 7 u g/ cm2, furathiocarb 1 15.7ug/ cm 2, 2 7.4ug/ cm 2, 3 4.3ug/ cm 2, 5 1.9ug/ cm 2, 7 1.1ug/ cm 2. - 17 -
3. Methid athion d islod geable resid ue d issipation. 4. Fu rathiocarb d islod geable resid u e d issip ation. - 18 -
(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 : 1344.1 u g/ hr DAT2 : 828.6 u g/ hr Calcu lation of reentry in terval DFR0 = 223 ng/ cm 2 K = -0.4954 ng/ cm 2/ d ay TF = 4872 cm 2/ hr Available d ata ADDSL = 1000 ng/ kg/ d ay (=ADI) BW = 60 kg ABS = 1.249 % So. Reentry Interval T=[ln{(1000 60)/(4872 0.01249 8)}-ln(223)] (-0.4954)-1 =1.2 day ( 28 hours ) - 19 -
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 : 204.7 u g/ hr DAT2 : 153.8 u g/ hr Calculation of reentry in terval DFR0 = 35.3 ng/ cm 2 K = -0.4138 ng/ cm 2/ d ay TF = 5116.2 cm 2/ h r Available d ata ADDSL = 3500 ng/ kg/ d ay (=ADI) BW = 60 kg ABS = 20 % So. Reentry Interval T=[ln{(3500 60)/(5116.2 0.2 8)}-ln(35.3)] (-0.4138)-1 = 0.7 day(18 hours) Fu rathiocarb REI d ata Carbofu ran. Carbofu ran REI48.. - 20 -
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 432.5 5621.9 ug/ cm2 8.65 g/ h 6110.7 patch 262.2 3408.6 ug/ cm2 5.244 g/ h 3705 patch 13.9 30.6 ug/ cm2 0.278 g/ h 33.3 patch ug/cm2 g/h patch 32.2 2287.6 ug/ cm2 0.644 g/ h 2486.5 patch 96.6 6856.5 ug/ cm2 1.9314 g/ h 7452.7 patch 421.2 24513.8 ug/ cm2 8.42 g/ h 26645 patch 292.8 7085.8 ug/ cm2 5.856 g/ h 7701.9 patch 401.4 30667.0 ug/ cm2 8.028 g/ h 33334 patch 505.8 6575.4 ug/ cm2 2.763 g/ h 7147.2 155.4 ug/ cm2 0.118 g/ h 168.9 4316.1 ug/ cm2 5.26 g/ h 4691.4-21 -
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 4.23 1.4 7.8 1.3 55.0 18.3 23.8 17.2 ug/ cm2 0.085 0.028 0.037 0.026 g/ h 14.7 5.2 6.6 4.9 patch 0.9 2.73 1.2 3.0 11.7 35.5 15.2 38.6 ug/ cm2 0.018 0.055 0.023 0.059 g/ h 3.12 10.1 4.2 10.9 patch 0.27 0.8 0.8 0.5 0.6 1.7 1.7 1.1 ug/ cm2 0.0054 0.015 0.016 0.010 g/ h 0.16 0.5 0.5 0.3 patch ug/cm2 g/h patch 1.77 0.9 1.6 0.5 125.7 63.9 115.0 38.3 ug/ cm2 0.035 0.018 0.032 0.011 g/ h 33.5 18.3 31.8 10.9 patch 0.96 0.5 1.1 0.9 68.2 38.3 74.6 61.8 ug/ cm2 0.019 0.011 0.021 0.017 g/ h 18.2 11.0 20.6 17.5 patch 2.64 4.1 3.5 2.9 153.6 237.5 202.5 171.1 ug/ cm2 0.053 0.082 0.070 0.059 g/ h 41.0 67.8 55.9 48.5 patch 7.62 6.5 9.9 10.3 184.4 156.8 239.6 249.7 ug/ cm2 0.152 0.130 0.198 0.206 g/ h 49.2 44.8 66.2 70.7 patch 9.54 2.2 4.7 1.8 728.9 165.0 362.1 137.5 ug/ cm2 0.191 0.043 0.095 0.036 g/ h 194.4 47.2 100.0 39.0 patch 4.14 2.7 5.3 1.7 53.8 35.1 68.6 21.8 ug/ cm2 0.023 0.015 0.029 0.010 g/ h 14.4 10.0 19.0 6.2 124.8 173.7 22.2 21.9 ug/ cm2 0.095 0.123 0.017 0.017 g/ h 33.3 49.6 6.1 6.2 9164.1 5375.1 368.7 353.1 ug/ cm2 11.176 6.555 0.450 0.431 g/ h 2443.8 1535.7 101.9 100.0-22 -
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 0.6 1.3 0.8 5.2 7.8 16.8 10.1 67.1 ug/ cm2 0.012 0.026 0.016 0.103 g/ h 2.0 4.1 2.6 16.3 patch 0.3 0.5 0.8 0.5 3.9 6.2 10.5 6.6 ug/ cm2 0.006 0.010 0.016 0.010 g/ h 1.0 1.5 2.7 1.6 patch ND 1.4 0.5 0.4 ND 3.0 1.1 0.8 ug/ cm2 ND 0.028 0.016 0.007 g/ h ND 0.7 0.3 0.2 patch 0.3 0.8 0.5 0.5 0.9 2.3 1.5 1.4 ug/ cm2 0.001 0.004 0.010 0.002 g/ h 0.2 0.6 0.4 0.3 patch 0.12 0.2 ND 1.4 8.5 14.9 ND 102.2 ug/ cm2 0.002 0.004 ND 0.029 g/ h 2.1 3.6 ND 24.8 patch 0.12 0.3 ND 0.4 8.5 21.3 ND 27.7 ug/ cm2 0.002 0.006 ND 0.008 g/ h 2.1 5.2 ND 6.7 patch 1.2 3.3 4.2 2.3 69.8 192.1 244.4 136.2 ug/ cm2 0.024 0.066 0.084 0.047 g/ h 17.5 46.6 63.5 33.1 patch 4.2 5.0 4.3 6.4 101.6 120.5 104.5 155.4 ug/ cm2 0.084 0.100 0.086 0.128 g/ h 25.4 29.3 27.2 37.7 patch 0.3 4.3 1.3 3.8 22.9 325.5 100.8 288.8 ug/ cm2 0.006 0.085 0.026 0.076 g/ h 5.7 79.0 26.2 70.1 patch ND 2.5 1.0 2.9 ND 32.0 12.5 38.2 ug/ cm2 ND 0.013 0.005 0.016 g/ h ND 7.8 3.2 9.3 19.5 135.0 7.8 148.5 ug/ cm2 0.015 0.103 0.006 0.113 g/ h 4.9 32.8 2.0 36.0 176.7 5607.0 425.7 4789.2 ug/ cm2 0.216 6.838 0.519 5.841 g/ h 44.2 1360.9 110.6 1162.4-23 -
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 5.25 6.2 ND 8.0 68.3 80.0 ND 103.5 ug/ cm2 0.105 0.123 ND 0.159 g/ h 18.5 21.2 ND 27.6 patch ND ND 5.4 ND ND ND 70.5 ND ug/ cm2 ND ND 0.109 ND g/ h ND ND 19.3 ND patch ND 5.4 5.4 ND ND 11.8 11.9 ND ug/ cm2 ND 0.107 0.108 ND g/ h ND 3.1 3.3 ND ND ND ND ND patch ND 5.4 ND ND ND 385.5 ND ND ug/ cm2 ND 0.109 ND ND g/ h ND 102.3 ND ND ND ND ND ND patch 4.26 10.4 12.9 10.5 247.9 607.6 749.6 611.8 ug/ cm2 0.085 0.209 0.258 0.210 g/ h 67.4 161.2 205.4 163.2 patch 12.54 12.4 21.4 12.2 303.5 300.6 517.6 295.5 ug/ cm2 0.251 0.248 0.428 0.0244 g/ h 82.5 79.7 141.8 78.8 patch ND 15.7 11.2 14.1 ND 1201.0 857.9 1079.0 ug/ cm2 ND 0.314 0.225 0.283 g/ h ND 318.6 235.0 287.7 patch ND 17.4 10.3 12.8 ND 226.2 134.3 166.5 ug/ cm2 ND 0.095 0.056 0.07 g/ h ND 60 36.8 44.4 ND 126.9 ND 146.2 ug/ cm2 ND 0.097 ND 0.112 g/ h ND 33.7 ND 39.0 289.5 2010 451.0 2574.9 ug/ cm2 0.353 2.451 0.550 3.140 g/ h 78.7 533.2 123.6 686.7-24 -
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 10.4 4.0 16.27 3.5 ug/ cm2 0.008 0.003 0.124 0.003 g/ h 2.6 1.0 38.7 0.8 31.1 577.0 ND 560.9 ug/ cm2 0.038 0.704 ND 0.684 g/ h 7.7 137.1 ND 132.9-25 -
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 0.007 g/ h ND ND ND 6.9 ND ND ND ND patch ND ND 1.9 ND ND ND 108.3 ND ug/ cm2 ND ND 0.037 ND g/ h ND ND 26.9 ND ND ND ND ND ND ND ND ND ND ND ND ND ND 39.3 42 ND ug/ cm2 ND 0.037 0.032 ND g/ h ND 11.9 10.4 ND ND 157.5 23.1 153 ug/ cm2 ND 0.192 0.028 0.187 g/ h ND 39.2 5.7 38.0-26 -
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 78.5 1019.9 ug/ cm2 1.569 g/ h 1108.5 patch 133.3 1732.4 ug/ cm2 2.665 g/ h 1883 patch ND ND ug/ cm2 ND g/h ND patch 5.9 17.6 ug/ cm2 0.27 g/ h 19.1 patch 11.16 792.4 ug/ cm2 0.223 g/ h 861.3 patch 173.2 12298.6 ug/ cm2 3.464 g/ h 13368 patch 169.2 9889.3 ug/ cm2 3.398 g/ h 10749 patch 238.6 5774.6 ug/ cm2 4.772 g/ h 6276.7 patch 48.1 3676.4 ug/ cm2 0.962 g/ h 3996.1 patch 91.7 111.8 ug/ cm2 0.501 g/ h 1295.5 760.8 ug/ cm2 0.561 g/ h 827.0 1118.7 ug/ cm2 1.364 g/ h 1216-27 -
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 0.092 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 1534.0 ND ND ug/ cm2 ND 0.529 ND ND g/ h ND 391.9 ND ND ND ND ND ND patch ND 5.4 9.3 ND ND 410.7 710.5 ND ug/ cm2 ND 0.108 0.186 ND g/ h ND 104.5 173.3 ND patch ND ND 26.5 ND ND ND 344.8 ND ug/ cm2 ND ND 0.145 ND g/ h ND ND 84.1 ND ND 45.6 10.2 ND ug/ cm2 ND 0.035 0.008 ND g/ h ND 11.6 2.5 ND 72.3 73.8 20.4 185.7 ug/ cm2 0.088 0.09 0.025 0.226 g/ h 17.9 18.8 5.0 46.7-28 -
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 3.2 3.4 7.9 ND 41.0 44.1 102.6 ND ug/ cm2 0.063 0.068 0.158 ND g/ h 9.6 10.1 23.7 ND patch 3 ND 29.2 ND 39 ND 379.9 ND ug/ cm2 0.06 ND 0.584 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 0.073 ND 0.083 g/ h ND 1.8 ND 2.1 patch ND 3.9 2.0 3.5 ND 11.8 6.0 10.5 ug/ cm2 ND 0.018 0.009 0.016 g/ h ND 2.7 1.4 2.4 ND ND ND ND ND ND ND ND patch ND 19.5 ND 7.9 ND 1134.9 ND 457.5 ug/ cm2 ND 0.39 ND 0.157 g/ h ND 260.9 ND 104.7 patch ND 6.4 ND ND ND 153.9 ND ND ug/ cm2 ND 0.127 ND ND g/ h ND 35.4 ND ND ND ND ND ND patch ND 8.9 ND 8.0 ND 115.4 ND 103.7 ug/ cm2 ND 0.049 ND 0.04 g/ h ND 26.5 ND 23.7 ND 57.3 ND 39.3 ug/ cm2 ND 0.044 ND 0.03 g/ h ND 13.2 ND 9.0 54 164.7 ND ND ug/ cm2 0.066 0.201 ND ND g/ h 12.6 37.9 ND ND - 29 -
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 17.4 27.3 66.6 69.0 ug/ cm2 0.021 0.033 0.081 0.084 g/ h 3.6 5.8 14.0 13.9-30 -
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 3.9 25.2 81.3 119.4 ug/ cm2 0.005 0.031 0.099 0.146 g/ h 0.9 6.0 19.4 28.4-31 -
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 30 27.3 ug/ cm2 ND ND 0.037 0.033 g/ h ND ND 7.5 6.6-32 -
30. m ethid athion furathiocarb (surfactant ) Methidathion Furathiocarb u g/ 405cm 2 10-3 u g/ cm 2 Mean u g/ 405cm 2 10-3 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 94.0 232.1 15.7 38.7 69.7 172.0 222.6 46.5 15.5 38.2 106.7 263.5 11.8 29.0 77.7 192.0 4.1 10.2 66.1 163.2 146.3 56.0 5.6 13.9 33.9 83.8 5.6 16.3 22.9 56.6 4.0 9.8 45.0 111.1 82.6 27.3 4.8 11.9 32.4 80.0 3.5 8.7 13.2 32.7 2.9 7.1 8.8 21.7 29.0 6.3 1.6 3.9 13.2 32.5 1.4 3.3 4.3 10.5 1.4 3.6 4.5 11.1 12.1 2.2 ND ND 5.9 14.6 0.6 1.6 N D 35.3 5.4 13.5 3.1 10.1 1.6 4.8 2.0 2.6 1.4-33 -
31. Methid athion furathiocarb (acetone ) Control 1 Day 2 Day 3 Day 5 Day 7 Day M ethid ath ion u g/ 405cm2 10-3 u g/ cm 2 N D N D Fu rathiocarb Mean u g/ 405cm 2 10-3 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 6.3 15.6 7.9 19.5 5.2 12.8 14.2 1.9 5.3 13.1 6.7 16.5 5.9 14.6 3.9 9.6 6.0 14.8 12.4 5.7 5.1 12.6 2.4 5.9 0 0 4.3 10.6 3.7 9.1 2.8 6.9 8.3 2.0 0.7 1.7 3.0 7.4 0.8 2.0 1.0 2.5 0 0 0.7 1.7 2.1 0.4 0 0 0.9 2.2 2.3 5.7 0.5 1.2 0 0 0.5 1.2 0.8 0.7 0 0 0 0 1.3 3.2 Mean S.D. N D 15.7 3.4 7.4 6.6 4.3 4.2 1.9 0.33 1.1 1.8-34 -
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 - 35 -
2) Biological monitoring (1),, A. a) Methid athion b) Furathiocarb B. Methid athion Furathiocarb 302.3 382.5 C6H 11N 2O4PS3 C18H 26N 2O5S colou rless crysta yellow viscous liqu i 39-40 - - > 250 KOW, logp 2.2 4.6(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) - 36 -
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(96 0.01mg/ L 0.002m g/ L 0.014m g/ L - - - - - 0.18ug/ L 0.03-0.12m g/ - 37 -
(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 (300-350g). 23 2, 50 10%, 12(7-7 ), cage 3-4. C. Animal clip per : Oster Pro-power 606-23E(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) - 38 -
(3) 3-1. 3-1-1. SD rat ether,.,.,.. (a-cyanoacrylate), (, 2cm )., (0.9% saline solu tion) 18m L glass bead. 32 1 incubation,. 3-1-2., 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,. 3-1-3., acetone 200uL m ethid athion 20, 50, 100, 200mg. (250rpm ) (2, 4, 6, 8, 12, 18, 24-39 -
) 500u L 500u L. m embrane filter, H PLC au tosamp ler 10uL chromatogram p eak methid athion,. 3-1-4. 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,. 3-1-5. 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,. 3-2. 3-2-1. 20, 50, 100, 200, 500uLmethid athion (Figure 5), 20u L100uL - 40 -
m ethid athion, 100u L. 20u L 50uL., 100uL,, 200uL. 3-2-2. 20, 50, 100, 200mg methid athion (Figure 6), 20m g, 50mg. 50mg m ethid athion. 100mg. - 41 -
3-2-3. 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, 24. 33. Methid athion (: pp m) 2 4 6 8 12 18 24 36 48 8.68 15.42 21.88 28.96 39.34 45.97 65.49 76.79 100.60 0.88 0.86 1.94 1.65 1.63 1.67 1.04 4.27 16.85 6.85 14.73 22.33 29.40 41.08 57.28 69.43 85.50 99.63 0.87 1.59 3.19 3.51 4.37 7.75 8.13 8.53 14.05-42 -
- 43 -
3-2-5. 1) 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 3-2-6. (LOD, limit of detection) (LOQ, limit of quantification) 1) Methid athion (LOD) : 0.03pp m(0.3ng) (LOQ) : 0.1p pm (1ng) 3-2-7. 1) Methid athion Methid athion m ethanol 1, 2, 5, 10, 20, 50, 100p pm, 10u L H PLC chrom atogram p eak (Figure 9). - 44 -
(4) Biological m onitoring I 4-1. Methid athion 4-1-1. 4-1-1-1. 5 x 6cm 2.5 x 2.5cm LD50 3/ 4, 3/ 20, 3/ 40 1159.5mg/ kg, 231.9m g/ kg, 115.95mg/ kg. metabolic cage2, 4, 6, 8, 12, 24 48,,,. 4-1-1-2. 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 - 45 -
. 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 ). - 46 -
4-1-2. 4-1-2-1. m ethid athion m ethid athion LD50 3/ 4, 3/ 20, 3/ 40, 12, 8, 6 ( 34, Figure 10). 34. Methidathion (: ppm) 2 4 6 8 12 24 48 LD50, 0.16 0.19 0.22 0.25 0.32 0.25 0.09 3/ 4 RSD 3.53 26.96 7.87 16.43 4.82 4.68 0.00 LD50, 3/ 20 LD50, 3/ 40 0.05 0.08 0.09 0.12 0.11 0.06 0.04 RSD 21.65 13.86 26.65 24.74 20.38 0.00 56.77-0.03 0.10 0.06 0.05 0.03 0.01 RSD - 17.32 10.00 10.19 12.37 17.32 5.09-47 -
4-1-2-2. methid athion methid athion m ethid athion ( 35, Figu re 11). 35. Methid athion (: u g) 2 4 6 8 12 24 48 LD50, 0.32 0.47 1.75 2.43 3.42 4.63 21.25 3/ 4 RSD 1.82 92.92 29.82 57.83 82.20 72.70 9.42 LD50, 3/ 20 LD50, 3/ 40 0.16 0.72 0.75 0.87 1.24 3.15 12.09 RSD 39.36 44.18 71.65 57.32 60.20 27.40 40.01 0.11 0.11 0.26 0.53 0.62 0.85 1.94 RSD 9.09 18.18 21.46 34.89 31.49 2.97 66.85-48 -
4-1-2-3. 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) 2 4 6 8 12 24 48 0.0034 0.0053 0.0141 0.0557 0.0338 0.0073 (16.64) (18.68) (9.03) (21.07) 0.05 0.05 0.14 0.80 (14.14) (14.14) (25.25) (7.95) (24.27) (28.28) 0.47 0.07 (13.54) (10.10) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 49 -
4-1-2-4. 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) 2 4 6 8 12 24 48 0.23 0.27 0.34 0.40 0.35 0.22 0.07 (17.49) (21.04) (8.37) (2.76) 0.65 0.86 0.88 1.18 (21.76) (5.79) (13.51) (3.81) (5.06) (17.10) (5.44) 0.92 0.56 0.21 (6.96) (27.78) (16.50) - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 50 -
4-1-3. 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 4-1-4. (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) 4-1-5. Methid athion hexane 0.02, 0.05, 0.1, 0.2, 1, 2pp m, 5uL GC chrom atogram p eak (Figure 14). - 51 -
4-1-6. 1),, 500 ul methid athion 0.05, 0.5, 2pp m,. m ethid athion 88.87-92.34% 95.36-105.96% ( 38). 38. m ethid athion (pp m) (%) 1 2 3 RSD(, %) 0.05 90.80 92.40 83.40 88.87 5.40 0.5 88.33 92.67 87.97 89.66 2.92 2 91.63 91.20 94.19 92.34 1.75 0.05 104.2 109.47 104.2 105.96 2.87 0.5 99.33 96.78 93.23 96.45 3.18 2 98.53 89.64 97.90 95.36 5.20 2),, 400m g methid athion 0.02, 0.05, 0.1pp m, 0.05, 0.1, 0.5p pm,. methid athion 84.0-94.7% 83.2-95.4% ( 39). 39. m ethid athion (p pm ) (%) 1 2 3 RSD(, %) 0.02 84.2 84.2 94.7 87.7 6.06 0.05 86.0 86.0 84.0 85.33 1.35 0.1 90.0 91.3 91.3 90.87 0.83 0.05 85.2 88.4 83.2 85.6 3.06 0.1 90.1 89.4 92.4 90.63 1.73 0.5 95.4 92.3 88.3 92.0 3.87-52 -
4-2. Furathiocarb 4-2-1. 4-2-1-1. 5 x 6cm 2.5 x 2.5cm 1500.0m g/ kg, 300.0mg/ kg, 150.0mg/ kg. metabolic cage2, 4, 6, 8, 12, 24 48,,,. 4-2-1-2. Furathiocarb 1) 700uL 1.5mL Ep pend orf tu be, (EtOAc:H exane=75:25) 700u L30 2 vortexing, 4, 20,000 rpm 10. 500u 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 10. 500u L N 2, methanol 125uL 0.45u m m embrane filter H PLC autosamp ler 10u L chromatogram peak furathiocarb, furathiocarb. - 53 -
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. - 54 -
4-2-2. 4-2-2-1. 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* 2 4 6 8 12 24 48 0.020 0.025 0.019 0.046 0.036 0.059 0.030 RSD 30.17 7.39 20.86 10.86 24.27 39.24 7.93 0.035 0.043 0.033 0.080 0.062 0.102 0.052 RSD 30.17 7.39 20.86 10.86 24.27 39.24 7.93 0.051 0.071 0.133 0.208 0.069 0.119 0.132 RSD 11.74 5.89 6.28 6.17 4.41 14.13 19.70 0.021 0.029 0.038 0.063 0.039 0.048 0.033 RSD 31.23 44.83 15.56 21.03 1.49 4.78 43.71 0.122 0.169 0.290 0.461 0.181 0.282 0.281 RSD 10.22 16.22 1.72 4.26 2.61 11.38 24.20 *) Total FC : CF 3-H C FC. - 55 -
- 56 -
4-2-2-2. furathiocarb furathiocarb carbofuran 3-hydroxycarbofuran, furathiocarb ( 40, Figu re 17, 18, 19). 40. Fu rathiocarb (: u g) CF 150 3-HC mg/ kg Total FC* CF 300 3-HC mg/ kg Total FC* CF 1,500 3-HC mg/ kg Total FC* 2 4 6 8 12 24 48 0.19 0.21 0.35 0.52 2.09 3.40 4.87 RSD 27.85 42.32 14.29 20.15 24.71 10.73 3.08 0.12 0.21 0.42 0.80 1.22 3.91 9.63 RSD 30.10 35.18 33.60 23.49 11.91 20.58 57.29 0.52 0.71 1.29 2.17 5.58 12.18 23.93 RSD 8.38 32.77 24.21 7.44 20.29 14.52 36.06 0.12 0.17 0.82 0.72 0.98 4.39 4.93 RSD 22.51 11.85 27.81 29.14 20.00 45.46 45.41 0.07 0.20 0.33 0.42 3.25 13.47 29.64 RSD 10.78 26.95 10.20 65.19 48.16 36.17 19.59 0.29 0.70 1.96 2.57 6.94 39.10 56.01 RSD 12.81 13.76 21.02 24.24 41.22 18.03 9.00 0.16 0.23 2.50 4.88 2.64 13.57 18.89 RSD 9.75 37.15 2.55 13.91 32.74 34.24 42.71 0.12 0.52 1.47 4.14 7.14 23.44 61.30 RSD 36.56 35.93 10.58 30.92 29.70 28.29 7.56 0.47 1.23 6.69 15.09 16.06 61.25 131.47 RSD 20.37 36.12 2.01 18.87 13.09 10.48 12.18 *) Total FC : CF 3-H C FC. - 57 -
- 58 -
- 59 -
4-2-2-3. fu rathiocarb fu rathiocarb 8 ( 41, Figu re 20) 41. Fu rathiocarb CF 1,500 3-HC mg/ kg Total FC* 2 4 6 8 12 24 48 0.069 0.109 0.152 0.824 0.977 1.194 0.456 RSD 52.65 18.90 47.14 12.01 23.39 7.41 25.30 0.00 0.087 0.105 0.329 0.293 0.278 0.109 RSD 0.00 58.04 5.39 21.49 39.10 82.41 12.38 0.118 0.327 0.431 1.955 2.160 2.418 0.963 RSD 52.73 35.46 26.58 2.93 26.84 14.42 18.44 *) Total FC : CF 3-H C FC. - 60 -
4-2-2-4. furathiocarb furathiocarb 8 ( 42, Figure 21) 42. Fu rathiocarb CF 1,500 3-HC mg/ kg Total FC* 2 4 6 8 12 24 48 0.00 0.259 0.517 1.918 0.388 1.292 0.347 RSD 0.00 10.04 28.45 20.47 34.28 38.41 14.20 0.00 0.00 0.00 0.00 0.00 0.00 0.00 RSD 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.448 0.893 3.315 0.671 2.233 0.599 RSD 0.00 10.05 28.44 20.46 34.28 38.41 14.22 *) Total FC : CF 3-H C FC. - 61 -
4-2-3. 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) 4-2-4. (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) - 62 -
4-2-5. 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). 4-2-6. 1),, 700 ul furathiocarb 0.5, 1p pm,. 2), 500m g 1g furathiocarb 0.5pp m 1p pm,. - 63 -
43. fu rathiocarb (%) RSD( (ppm) 1 2 3, %) 3-HC 0.125p p m 92.6 92.8 89.0 91.47 2.34 0.5p p m 99.65 76.6 95.95 90.73 13.64 3-KC 0.125p p m 100.8 100.4 93.6 98.27 4.12 0.5p p m 103.0 99.5 88.5 97.00 7.80 CF 0.125p p m 103.8 108.0 97.0 103.13 5.65 0.5p p m 112.6 110.0 100.0 107.5 6.20 FC 0.125p p m 90.6 93.2 94.0 92.6 1.92 0.5p p m 94.9 96.9 97.0 96.25 1.22 3-HC 0.125p p m 93.6 98.8 88.2 93.53 5.67 0.5p p m 70.9 77.9 67.4 72.07 7.42 3-KC 0.125p p m 89.2 96.2 91.0 92.13 3.95 0.5p p m 88.95 95.25 84.95 89.72 5.79 CF 0.125p p m 100.4 95.8 101.0 99.07 2.87 0.5p p m 87.0 86.05 75.6 82.88 7.63 FC 0.125p p m 95.6 95.4 86.4 92.47 5.68 0.5p p m 76.3 77.85 76.65 76.93 1.06 44. furathiocarb (%) RSD( (pp m) 1 2 3, %) 3-H C 0.5p p m 83.6 87.6 85.5 85.57 2.34 1.0p p m 85.7 88.3 88.6 87.53 1.82 3-KC 0.5p p m 89.4 90.0 90.6 90.00 0.67 1.0p p m 91.7 87.4 91.25 90.12 2.62 CF 0.5p p m 90.6 92.6 88.2 90.47 2.44 1.0p p m 92.5 94.7 93.85 93.68 1.18 FC 0.5p p m 86.0 82.6 84.3 84.30 2.02 1.0p p m 84.6 84.6 87.5 85.55 1.92 3-H C 0.5p p m 84.2 78.6 79.8 80.87 3.65 1.0p p m 81.7 85.1 87.6 84.80 3.49 3-KC 0.5p p m 87.6 89.8 85.6 87.67 2.40 1.0p p m 90.6 91.2 91.7 91.17 0.60 CF 0.5p p m 89.6 95.2 91.8 92.20 3.06 1.0p p m 92.9 96.9 98.1 95.97 2.84 FC 0.5p p m 85.6 87.2 89.2 87.33 2.07 1.0p p m 88.5 90.4 93.1 90.67 2.55-64 -
(5) : 5-1.. :,. PBPK.. 5-2. 5-2-1. Methid athion 5-2-1-1. Methid athion 22m g 1mL DMSO 100mL. 220pp m m ethid athion 1% DMSO. 0 24 H PLC, m ethid athion. 5-2-1-2. (,, ) g3ml hom ogenizer. 5-2-1-3. 1mL220pp m methid athion 100u L0 1, 2, 4, 6, 8, 10. Methid athion 30,000g, 4 20 0.45u m mem brane filter m ethid athion H PLC methid athion. - 65 -
5-2-1-4.,, 1mL220p pm m ethid athion 100uL0 5. methid athion 20pp mdmso 0.1%. Methid athion 30,000g, 4 20 0.45u 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. 5-3. 5-3-1. Methid athion 5-3-1-1. 0 220p pm methid athion, 0-24 methid athion. m ethid athion. - 66 -
5-3-1-2. Figure 23 m ethid athion 4. :, : 5. 5. - 67 -
5-3-1-3. Methid athon : 8. 9: 9. 45. methidathion : (SD) Tissu e Ct Cw Ptw = Ct/ Cw (p pm RSD (pp m RSD RSD Liver 78.79 0.99 5.30 3.68 14.87 4.76 Kid ney 57.57 1.35 7.78 3.26 7.41 4.53 Blood 71.86 3.88 13.65 2.50 5.27 6.41 46. Tissue:blood p artition coefficients Tissu e P = Ptw / Pbw Liver, PL 2.82 Kid n ey, PK 1.41-68 -
(6) Biological m onitoring II - Field test 6-1. Methid athion 6-1-1. 6-1-1-1. 6-1-1-2. 6-1-1-3. 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 3 2. 50mL, sod iu m su lfate 4 0. H exane 1mL GC au tosam pler hexane 5uL chromatogram p eak methid athion(50 ). - 69 -
6-1-2. 6-1-2-1. methid athion (: p pm ) N ame Date Comm ent Average(n=3) RSD 5/ 13 0.0096 2.08 5/ 14 0.0052 17.63 5/ 14 0.0074 7.64 5/ 12 Blan k 0.0000-5/ 12 0.0045 9.43 5/ 12 1 0.0105 6.73 5/ 13 0.0049 8.55 5/ 14 0.0054 20.62 5/ 14 0.0076 6.96 5/ 13 0.0049 8.44 5/ 14 0.0051 18.41 5/ 14 0.0043 13.53 5/ 13 0.0120 7.26 5/ 14 0.0057 23.23 5/ 14 0.0136 10.43 6-1-2-2. methid athion N am e Date Time Com ment urine(m L) concentration qu antity (ppm ) (ug) RSD 5/ 13 13:30 1, 150 0.0016 0.24 11.92 5/ 14 7:30 2, 250 0.0002 0.06 11.79 21:00 2, 210 0.0010 0.2016 5.51 5/ 15 8:15 3, 260 0.0003 0.0702 15.71 5/ 13 19:00 1, 280 0.0003 0.0784 10.10 7:00 2, 330 0.0000 0.0000-5/ 14 12:44 2, 290 0.0003 0.0841 24.38 18:00 2, 220 0.0000 0.0000-5/ 15 7:30 3, 250 0.0000 0.0000-5/ 13 19:00 1, 240 0.0006 0.1504 13.29 5/ 14 6:50 2, 120 0.0020 0.2436 6.27 9:50 2, 380 0.0007 0.2584 5.09 5/ 15 7:30 3, 120 0.0005 0.0588 25.98 5/ 13 19:00 1, 270 0.0013 0.3636 8.70 5/ 14 5:30 2, 300 0.0002 0.06 11.79 19:00 2, 360 0.0007 0.2616 11.12 5/ 15 6:00 3, 350 0.0005 0.1855 2.67-70 -
6-1-2-3. (LOD, limit of detection) (LOQ, limit of quantification) (LOD) (LOQ) 0.002p pm (0.01ng 0.0002p pm (0.001ng 0.006p pm (0.03ng 0.0006p pm (0.003ng 6-2. Furathiocarb 6-2-1. 6-2-1-1. 6-2-1-2. 6-2-1-3. Furathiocarb 1) 700uL 1.5m L Ep pend orf tu be, (EtOAc:H exane=75:25) 700uL30 2 vortexing, 4, 20,000 rpm 10. 500u 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 3 2. 50mL, sod iu m su lfate 40. Methanol 200uL H PLC au tosamp ler 10u L chrom atogram peak fu rathiocarb (50 ). - 71 -
6-2-2. 6-2-2-1. furathiocarb (: p pm ), fu rathiocarb. N ame Date Com ment Concentration(ppm < 0.0025 6/ 2 < 0.0025 1 < 0.0025 6/ 3 < 0.0025 6/ 4 < 0.0025 < 0.0025 6/ 3 < 0.0025 < 0.0025 6/ 4 < 0.0025 6/ 3 < 0.0025 < 0.0025 6/ 4 < 0.0025 6/ 3 < 0.0025 < 0.0025 6/ 4 < 0.0025-72 -
6-2-2-2. 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 < 0.002-6/ 3 7:00 1, 340 < 0.002-6/ 3 16:00 1, 110 0.018 1.97 6/ 3 21:00 1, 250 0.013 3.29 6/ 4 7:00 2, 390 < 0.002-6/ 4 16:30 2, 240 < 0.002-6/ 4 22:00 2, 260 < 0.002-6/ 5 7:00 3, 330 < 0.002-6/ 3 21:30 1, 140 0.005 0.63 6/ 4 7:00 2, 210 0.006 1.33 6/ 5 7:30 3, 220 < 0.002-6/ 3 19:40 1, 280 < 0.002-6/ 4 7:00 2, 410 0.007 2.80 6/ 4 7:10 2, 200 < 0.002-6/ 5 6:00 3, 460 < 0.002-6/ 4 5:45 2, 340 0.002 0.80 6/ 4 19:43 2, 210 < 0.002-6/ 5 6:25 3, 400 < 0.002-6-2-2-3. (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 - 73 -
(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, - 74 -
: Biological monitoring : : 96 99(3 ), 3, - ( 97 ) - ( 97 ) - biological m onitoring ( 98 ) - ( 98 ) - () - - 75 -
- 1: - 2: - 3: (, ),, - - < > biological m onitoring. - : SPF S.D. - : Methid athion, furathiocarb. - 3. -, - ChE - 76 -