Evaluation of LC/MS and LC/UVD(EPA-631) Methods in Determinationof Benomyl Concentration in Water Samples Benomyl

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(1)Journal of the Korean Chemical Society 2001, Vol. 45, No. 1 Printed in the Republic of Korea. LC/UVD(EPA-631) Benomyl

(2) . LC/MS. *

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(4) (2000. 9. 22 ). ‡. † ‡. Evaluation of LC/MS and LC/UVD(EPA-631) Methods in Determination of Benomyl Concentration in Water Samples Kee D. Kim*, Won Gyu Choi†, Yong Chan Seo‡, Byung Hwang Park‡, and Hye Kyung Choi‡ Department of Chemistry, Sangji University, Wonju 220-702, Korea † Department of Mining and Minerals Engineering, Sangji University, Wonju 220-702, Korea ‡ Department of Environmental Engineering, Sangji University, Wonju 220-702, Korea (Received September 22, 2000). . benomyl !"#$ %&' () *$ +, -.. /0123 benomyl4 5)6 carbendazim23 78 9 :;4 carbendazim< +, = >;.. 6 benomyl4 5) ? 1@A BC3 DE6 F, +GHI.. FJ KL carbendazim *$ LC/MS (TOF)3 :-28 M N O)P ) Q RS T= UV$ GWS TI.. XEA= LC/UVD;(EPA-631)P 6 1,600Y Z[28 spike \]* 80~118%3 ^_-.. benomyl. ABSTRACT. Benomyl, one of the known endocrine disrupting chemicals, was analyzed to understand the its fate in water. Benomyl analysis process in water sample include the following sequential steps. Hydrolysis of benomyl into carbendazim, solvent extraction, concentration, and the concentration of final carbendazim solution was determined by LC/MS (TOF). Recoveries of the spiked samples were good with the range of 80.6 to 118.6% and the MDL was 1,600 times lower than that of LC/UVD method. The use of LC/MS (TOF) successfully eliminated the positive error incurred by interferencing materials in the matrix.. . u &BHI.. Benomyl4 1 z{ Z 4 |23 }~I2 1987w vs o_sP ) ^mQ 5{ T= 53 3 jH hP A B !, +6 T .. s Pe += D+( 67) 3 +H T28 YE%& 3 +HI< ( , 1999) +Pe GK= hP e B !,* +) T.. 750. Methyl (1-butylcarbamoyl)-2-benzimidazolecarbamate. (benomyl)4 `ab cd acetylcholine esterase reversible inhibitor3e f!QR gGhB T= carbamate i 8 h j ? k, l, mn o%G3 pq &BHr T.. sPe = t> 50u +*5 &BHr T28( , 1999) vs Pe= 1979wx 1986wy t> 1. 2. 40.

(5) LC/UVD(EPA-631) Benomyl

(6) . LC/MS. 41. )K vs = bq Â4 K>< Ã¨3 8 ´123 Ä® carbendazim< diazomethane 23 methylation< 6 m*A ?P GC/MS3 :< A.. Benomyl< carbendazim23 )6 +,µP T ÅÆ ÇjA aÁq; j¿ Ã¨8, vs HPLC/UVD$ BA :; O) ¶· K +,µP T U!$ QRKÈ 65 T.. /9 m*$ nA GC/MS; carbendazim diazomethaneP A N-methylation4 1 ÉPe Ê ¸ ®ËH' C ~Ì 0Í Î3 M O;P `Ï{< Ð'5 ~Ì | &" .. P benomyl< carbendazim23 5)6 :µP T O) ¶·P A ÑÒ< ´d 8 M XEA$ ZE T= :; ÓQ Ã¨A "+.. ¯e 9 tÔPe= LC/MS(TOF)$ B6 O) ÑÒ< GW8 ÕKP XEA $ ZD*Ö -r O;< B6 as ×Pe benomyl *$ %&-.. 7. Fig. 1. Structure of benomyl and carbendazim. Benomyl rat LD50= 10 g/Kg(WHO, 1993) 23 C Z4 z{< or T2 r P= A 100 mg/Kg 23* A < r T.. benomyl & methyl 2-benzimidazole carbamate(carbendazim) z{4 benomyl P ) 28 benomylP ) = z {4 benomyl R &P ) R{J carbendazimP A c 3 ¡Hr T.. Benomyl4 P A B)*5 Zr C ¢+ 6 t#Pe* 123 +A carbendazim2 3 £¤ )H= |23 }~ T.(Fig. 1). )¥ *= ¦*, , , i § ¨23 }~ T .. ©, k¦Peo. r¦Pe, {# o.= ª' {Pe )5 «¬¤ ®.. pHP ¯° benomyl 0±'= pH 3, 7, 9 470, 180, 30 2 3 }~ T.. M² )P ¯° R{4 { carbendazim, ª'{ P= 3-butyl-2,4dioxo-s-triazino[1,2-a]benzimidazole(STB)5 3 R{ H= |23 }~ T28 pH 3Pe 8yPe )4 carbendazim.. Carbendazim4 C +A 3e pH 8 B³Pe ´d 8/ + µ< or T.. ²A m3 benomyl4 M 3 ¶·'o.= carbendazim§ .° 3 a H ¶·' ¸¹P benomyl P ) : 'o.= carbendazim23 5)Kº DE, +G +< W» +,:< A ? ¼23 ,< 6 !*$ ¡+r T.. ½· benomylP A :;4 ¾s vs EPA Method-631, /9 :; § j¿H T.. vs :;4 ³-³ DE;< B6 DE< A ? {Pe 16-24K>P À» carbendazim23 5)$ 6 HPLC/UVD3 :< ¤ H T.. /9Pe* a Áq= ³-³ DE;< B carbendazim23 5 3. 3,4. 1,5. 6. 7. 2001, Vol. 45, No. 1. .. :P &BJ ''= HPLC(Varian 9012Q), ,:'(Micromass LCT)Ø UV XE'(Varian 9050 UVD)$ &B-.. ª ÙÚ, ÙÚÛ4 !:B< &B-28 ÜÝK 4 Þ* 99% benomyl carbendazim< Aldrich &Pe Ô6 &B-.. M ¾ Kß4 àK < &B-r d 5á= Þ* 99.999% rÞ * 1à d$ &B-..

(7) . 1 L â ãáäP 500 mL KL$ å4 ? 7 mL 1+1 æ magnetic stirrer$ år 20K>< "¦Pe 0Kç.. 10 N NaOH$ 156 KL pH$ 7-8 +*3 %+ A ? 1 L ³ è'P år 80 mL methylene chloride$ 5A.. éê< 2> ®ë6 10>P À» ì í qH*Ö +A ? í< qA.. ( DE< 4\ 0îr, qJ m'í4 ïì êA ð ñ sodium sulfate3 ò®J drying column< n Kç ? Kunderna-Danish(K-D) F' 3 1 mLy FKç.. Column ó ãáä$ 6 ÙÚÛ3 ô4 ? d$ ¢å ´ õ5 Benomyl.

(8) . 42. 5 H*Ö FKç.. ´ FJ KL= 30 cmö3.9 mm ID µ bondapak(10 µ) column< &B 6 Õ(ÙÚÛ : =1:1)< 0.8 mL/min m¥23 ®ËK÷8 LC/MS(TOF)3 q, :-.. 6'e MSP ,4 splitter$ &B6 1/12 KL ß5*Ö %@-.. : ¦= electrospray;< &B-28 XE¦(m/z)4 192.19(M+H) ¦< SIM(selected ion monitoring)3 +,:28 ,¡+4 TOF(time of flight);23 -.. UVD3 :< S P= EPA-631;Pe ÇKA øù 254 nm ` ´ øù 286 nmP e :< 6 MDL(method detection limit)< Z DI.. GC/MS$ &BS = /9;P ú % û< M3 &B-2 methylation< (A diazomethane ê{K Aldrich& Diazald kit$ &B6 ê{-.. 1 mL. +. vs EPA-631P ü benomyl :4 KL 150 mL$ ý6 ´ 10 mL3 FK÷8 M¸ MDL4 8.7 µg/L +*3 }~ T .. 0ü /9;P ü 1 L KL$ ý6 ûr Kç ? ´ 0.2 mL BC$ å ´ õ$ N +8 M¸ MDL4 0.1 µg/L23 }~ T.. 9 "þPe= DEBC$ FKºÿ Î3 q$ (6 ´õ$ 1 mL3 -r MDL Ò a Áq + q{ ó G{< ïì r~6 KL = 500 mL$ &B-.. "G3 LC/UVD3 ¡+S MDL< 0.1 µg/Ly ZE TI. aÁq ? FJ KL carbendazim *= LC/ MS(TOF)$ &B6 m/z5 192.19(M+H) ¦ SIM chromatogram< &B6 :-.. Carbendazim *P 1.52(benomyl ,/carbendazim ,)$ A benomyl *P A.. X, h{ ? 50 µg/L carbendazim ÜÝB³< Gh6 X,j(calibration check)< -r C 10Ó KL$ ¡+A ?P carbendazim ÜÝB³< ¡+6 '' +{< '123 X-.. Warning level3= 2δ(ÜÝV), control level3= 3δ $ &B-r warning level< t¥ 2\ W control level< S X,< · h{6 K +. 1,5. Table 1. MDL Study for the Determination of Benomyl by LC/MS (TOF) Method Sample Number MDL1 MDL2 MDL3 MDL4 MDL5 MDL6 MDL7 Standard Deviation MDL. Carbendazim Area Concentration Count (g/L) 72.8 68.0 65.2 67.8 69.2 68.2 68.6 2.262. 0.026 0.024 0.022 0.023 0.024 0.024 0.024 00.0011 00.0035. Benomyl Concentration (g/L) 0.039 0.036 0.034 0.036 0.036 0.036 0.036 00.0017 00.0053. *0.0304 µg/L of standard carbendazim solution was used for MDL study.. L$ :-.. MDL ¡+4 carbendazim< 0.0304 µg/L *5 H*Ö spike A 7Ó KL$ ¡+6 ÜÝV$ Ôr 98% `Ï* Student t 3.14$ ÜÝ VP 6 -.. Table 1Pe o= Ø LC/MS(TOF)3 ¡+A benomyl MDL4 0.0053 µg/ L23 /9;P ) 20Y, vs EPA ;P 6= 1,600Y Z4 < or T.. Spike \]< ¡+' (6 KL(500 mL)P 0.1, 0.2, 0.3 µg carbendazim< 5A ? Õ/A a Áq +< W» \]< ¡+-.. M N 118, 80, 81% ^_A \]< or T.(Table 2). /9;P ¯° benomyl 5) carbendazim diazomethaneP A Ù 0Í4 m'1 É Pe Ê ¸ ®ËH' ~Ì 0Í23 ¡J.. " G3 carbendazim 10 mg/L ÜÝB³< diazomethane 23 Áq6 GC/MS3 jA N methyl carbendazim R{< SIM23= jS TI2 total ion ä3 M

(9) 23= M peak< jS < +* d, R{< } TI.. * Ô%: ¢5A 6² peak XEH= |23 o carbendazim4 0Í 4 GC columnPe )H8 methyl carbendazim4 1% 3 { H= |23 ¡J.. vs EPA-631;P ¯ LC/UVD3 ¡+S , KLPe carbendazim 4 retention time < o= peak XEHI2 ' O) m' Journal of the Korean Chemical Society.

(10) LC/UVD(EPA-631) Benomyl

(11) . 43. LC/MS. Table 2. The benomyl concentration in water samples. . . . . Carbendazim Benomyl Sample Recovery Concentration Concentration Number (%) (µg/L) (µg/L) w-1 w-2 w-3 w-4. 0.506 1.861 0.466 ND. 0.768 2.829 0.707 ND. w-5 w-5-s1 w-6 w-7 w-8 w-9 w-10. 0.029 0.516 0.143 ND 0.584 0.265 ND. 0.044 0.783 0.217 ND 0.888 0.402 ND. w-11 w-12. ND ND. ND ND. w-13. 0.290. 0.440. w-14. ND. ND. w-15. 0.153. 0.232. w-16 w-17. 0.157 0.039. 0.238 0.059. w-18 w-19. ND 0.094. ND 0.143. w-20. ND. ND. w-21 w-22. ND 0.054. ND 0.082. Carbendazim Benomyl Sample Recovery Concentration Concentration Number (%) (µg/L) (µg/L). . w-23 w-24 w-25 w-26. 0.078 0.019 0.066 0.101. 0.117 0.028 0.100 0.153. . w-27 w-27-s2 w-28 w-29 w-30 w-31 w-32. 0.024 0.346 0.018 0.079 0.634 0.018 ND. 0.035 0.526 0.026 0.119 0.962 0.026 ND. w-33 w-34. 0.096 ND. 0.146 ND. w-35. 0.076. 0.115. w-36. 0.212. 0.323. w-37. 0.089. 0.135. w-38 w-39. ND 0.076. ND 0.115. w-40 w-40-s3. 0.046 0.283. 0.070 0.430. 81.2. .

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(17). w-41. ND. ND. w-42 w-43. 0.678 0.042. 1.030 0.064. 80.5. 118.5. *1, 2, 3; 0.3, 0.2, 0.1 g of carbendazim was spiked to the 500 mL of samples for recovery test, respectively.. 4 3 .° m'P A ½< YGS .. Fig. 2 M APe o= Ø "G KL LC/UVD3 ¡+A chromatogram 4 peak #5 4 K>P À» = { < or T .° O)P A positive error 5{< 5r T< ' ¢Þ3 A base line ¢+{* or T.. 0ü Fig. 2 M BPe o= LC/MS(TOF) SIM23 ¡ +A N = ï O) ó ¢ÞP A ÑÒ < ïì GW6 +J base line ó 4 ±*$ or T.. P ¯ LC/MS(TOF)3 as 13Ó × 43Ó Pe benomyl*$ ¡+-28 0.02 µg/L = ¢XE(N.D.)3 ÁqA N 70% +*(30Ó ¼)5 benomylP ) UªI< } 2001, Vol. 45, No. 1. Fig. 2. Carbendazim chromatograms of w-29 sample taken by LC/UVD (A) and LC/MS (TOF) in SIM (B)..

(18) . 44. Table 3. The benomyl concentration in water samples determined by LC/MS (TOF) and LC/UVD Sample Number. Determined by LC/MS (TOF). Determined by LC/UVD. Carbendazim Benomyl Carbendazim Benomyl Concentration (µg/L) Concentration (µg/L) Concentration (µg/L) Concentration (µg/L). w-15 w-17 w-19 w-22 w-23 w-24 w-26 w-29. 0.153 0.039 0.094 0.054 0.078 0.019 0.101 0079. 0.232 0.059 0.143 0.083 0.117 0.028 0.153 0.119. 5 T.(Table 2). /01 Pe B 1 < = KL= I2 KLý K'= 9, 103e '> < r~S ¸ s × benomylP ) ¥123 Uª H .r RS T.. ( 43Ó KL benomyl µJ |23 j J KL 8Ó KL$ ñh(3 DE6 aÁq ? LC/UVDØ LC/MS(TOF)3 carbendazim< +, ? benomyl3 -.(Table 3). M N LC/UVD P ) N+J *= Õ/KL$ LC/MS(TOF)3 +,A N P ) ´ 420%+* UV$ o= |23 o LC/UVDPe= O)P A , error5 QRS T< } T.. UV= 0.03Pe 0.3 µg/L +*3e carbendazim * P ñ¤ 7rT.. |23 o UV= m' O)P A |< mDS T.. mg/L. P A benomyl :;4 /9 : ; vs EPA-631P ) § Z4 MDL< o6! aÁq;* >.= T .. vs EPA-631 LC/UVDP A +,: P= O)P A positive error5 T< } TI.. ²A N 3 D Ê ¸ +jA :< (6= LC/MS$ BA +{, +, : Ã LC/MS(TOF). 0.244 0.102 0.113 0.187 0.208 0.097 0.121 0.353. Differences between LC/ MS (TOF) & LC/UVD (%). 0.371 0.156 0.172 0.284 0.316 0.148 0.184 0.536. 59.3 160.4 20.3 242.9 168.2 423.3 19.7 350.3. < } T.. Æ" q BC@±c 5 # micro-HPLC$ Nê6 &BS Æ" G1r ÒJ :N $ Ä< T< |23 J.. 9 tÔ= 2000w* tÔØ As · +' tÔ(1999-2-309-005-5) /

(19) 23 ËHI28 P ±&A... 1. Liu, C. H.; Maltern, G.; Yu, X.; Rosenu, J. L. Agri. Food. Chem. 1990, 38, 167. 2. National Research Council, Regulating Pesticides in Food, The Delaney Paradox; National Academy Press: Washington, D.C., U.S.A., 1987; p 89. 3. Lim, J.; Miller, M. G. Toxicology and Applied Pharmacology 1997, 142, 401. 4. Mallat, E.; Barelo, D.; Tauler, R. Chromatographia 1997, 46, 342. 5. Cano, P.; Plaza, J.; Munoz-Delgado, L. J. Agric. Food Chem. 1987, 35, 144. 6. U.S. EPA Analysis of Water, Method-631, 1998. , (Speed 98), 7. 1999. 8. Choi, K.; Lee, H.; Lee, D.; Byun, W. Anal. Sci. and Tech. 1994, 7, 421. 9. Han, I.; Chai, J.; Lee, J.; Yeo, I. Anal. Sci. and Tech. 1994, 7, 395..

(20) . Journal of the Korean Chemical Society.

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