Liquid Liquid Extraction-GC/MS

전체 글

(1)Journal of the Korean Chemical Society 2003, Vol. 47, No. 6 Printed in the Republic of Korea. . . Liquid Liquid Extraction-GC/MS (BHA, BHT).

(2) (Fenvalerate, Allethrin) †‡ *

(3) , †.

(4) ‡ (2003. 8. 18

(5) ). Simultaneous Determination of Antioxidant(BHA, BHT) and Insecticide(Fenvalerate, Allethrin) by Liquid Liquid Extraction-GC/MS Jae-Hee Lee†,Jun-Hyun Bae‡, Jun-Gill Kang*, and Youn-Doo Kim Department of Chemistry, Chungnam National University, Daejeon 305-764, Korea † Korea Water Resources Corporation, Daejeon 305-730, Korea ‡ Department of Environmental Engineering, Anyang University, Kyungki-Do 430-714, Korea (Received August 18, 2003). . . LLE-GC/MS ! "#$% & ' ( ) *(+ ,-. /0

(6) 1%. 2 345. 6 7.8 & ' (% butylated hydroxy anisole(BHA) ) butylated hydroxy toluene (BHT)93, *(% allethrin ) fenvalerate9:5. ;<-=. />.8 ?@ (LOD)% 0.071~0.159 ng/ml9:AB, 9 C D;<EF%(%RSD)% 1.41~5.349:5. G, HG, I,G ) JKG

(7) L.8 9M NMO ?@$P QR5. NS -=T U, ?@ (LOD)% 0.051~0.132 ng/ml9:AB, 9 C D;<EF (%RSD)% 6.39~13.4% 9:5.. : , , ABSTRACT. A procedure based on liquid liquid extraction(LLE)-GC/MS has been developed for the simultaneous analysis of antioxidants and insecticides known as endocrine disrupters. The endocrine disrupters investigated in this study are butylated hydroxy anisole(BHA) and butylated hydroxy toluene(BHT), and the insecticides are allethrin and fenvalerate. The limit of detection(LOD) was 0.071~0.159 ng/ml with RSD of 1.41~5.34% for the standard sample. From water reservior of Han river, Geum river, Nakdong river and Sumjin river, these compounds were not detected. For the synthesized sample, the LOD is 0.051~0.132 ng/ml with RSD of 6.39~13.4%. Keywrods: Liquid Liquid Extraction, Antioxidant, Insecticide. . disrupters)7 87 9:/+, 234 ;<8=

(8) > ?@A 56BC'. 234 56D .

(9) . ;1 E7 FG H" IJ, K+LM NO,. !" #$%& '( )*#$%. , ,

(10) PQR, ST U VT estrogen. +(, -./( 01 234 56(endocrine. R, SWXN, polychlorinated dibenzo-p-dioxins(PCDDS),. . 559.

(11)

(12) . 560. polychlorinated dibenzo furans(PCDFS) Y& Z [\ ] ^_ `\ ab 0'. cd ]7 e fg(\N 0(,

(13) h i1 TD /? j8k, lm8 nT op qOrsD, = t uv wx

(14) yz ] { U 2| }/ ~7 '. , 234 56] 2|v , 1991 , w m Eichelberger Y7 "

(15) 2|1 liquid solid extraction-GC/MS "(, 1994, Pang Y7 &: R1 pyrethroid 2|1 capillary column-GC& packed column-GC 35?8"'.1-3 d, 1995 Munch 1 "

(16) 2|1 liquid solid extraction & capillary column-GC/MS "'.4 1997, Tena Y7 D\p 1 supercritical fluid extration(SFE) "( 1998 , Ramesh Y7

(17) h p pyrethroid 2|1 GC/electron capture detector(ECD) "'.5,6 2001, Mikael Eriksson Y7 |sD. Fig. 1. Chemical structures of BHA, BHT, allethrin and fenvalerate used in this study.. 9 O 2|1 solid phase microextraction(SPME)-GC/MS "(, 2002,. 8"»sD¼ 8VT ½ ²='. ¾ªD. Brito Y7 R1 2|1. " BHA, BHT, allethrin U fenvalerate µS7. liquid liquid extraction(LLE)-GC solid phase extraction. Fig. 1p 1 Z'.. (SPE)-HPLC "'. 2002, Brossa Y7 7,8. BJ 234 56 2| solid phase. . . extraction-GC/MS "(,9 Petrovic Y7 . \?sD1 liquid liquid extraction(LLE),3,11. BHA U BHT1 98% ¿, Wako W, \ ( allethrin U fenvalerate1 98% ¿, Dr. Ehrenstorfer. liquid solid extraction(LSE)4 U solid phase extraction. W ;/ ~( "'. "ÀD1 Wako. (SPE) Y . 0/¡,2,9,13 U 2. R¥ÁBÂ"= n-hexane U Merck HPLC. |1 e ¢zp LLE 9#d EsD Oe. Ã dichloromethane "'. NaCl7 :« Junsei. b 0'. £¤, large volume injection(LVI) d. GRÃ BÁ 450 oCp 4BÄ ÅBÆ

(18) . LLE7 ¥j¦§ d z¨ ©@ ª'.. Ad Ç " '.. )*BJ 234 56] 2|1 MS "d 0'.. 10. 3,11. « ¬p1 B 234 56 :s. BHA, BHT, allethrin U fenvalerateBÁ] n-hexane. ®1 D ;( 01 2G , butylated. sD È|, FF 4.4, 3.2, 3.2 U 2.9 µg/mLÉ . hydroxy anisole(BHA) U butylated hydroxy toluene(BHT),. µ'. ½ ¾ª"sD1 benzyl benzoate I. 2G , allethrin U fenvalerate LLE-GC/MS. (5,000 µg/ml in methanol) 50 µl J.T. Baker HPLC. sD B 2|¯ 01 ? e°( . Ã methanol 10 mLD È| 25 µg/mLÉ µ. '. ±+p, ] ²= U 2|1 ³1. '. ÊR1 Water purification system(Milipore,. :´ µ¶] 8 8 d ', « ?. USA) &BÆ 18.1 ΜΩ < ÊR À 24B. d·, P·, ¸· U ¹º· I VTBJ . Ä ' 5)k "'. Journal of the Korean Chemical Society.

(19) Liquid Liquid Extraction-GC/MS (BHA, BHT) (Fenvalerate, Allethrin) . 561. Table 1. Injector operating condition for large volume injection Time. Temp. (oC). Rate (oC/min). Split state. Split ratio. Column flow (ml/min). 0.00 0.75 3.00 20.000. 080 280 280 280. − 200 − −. on off on on. 100:1 off 20:1 20:1. 0.1 1.2 1.2 1.2. Gas Chromatograph1 Varian CP 3800 GCDp Ë , ÌD Í N>(, rb" injectorN Î Ïb0'.BJ;r^Ð@ Combi-Pal Autosampler (CTC analytics, Swiss) "'. Capillary column7 Varian CP Sil-8 CBD ÑN 30 m, * 0.25 mm '. Column (;<7 0.25 µm ÒÓ 5% diphenyl & 95% polydimethylsiloxanesD Ôb 3qT ( Õ Ö;k, ?v× V ØÙT 0'. column7 −50~320 oC ÚÐ Ë,p " N> (, "¯ y1 w\ 300 oCp 12BÄ < N (;< ÛÜ Ý,Þ '.. Table 3. Mass spectrometer operation parameters Parameter. Value. Ionization Mode. E.I (70 eV) auto gain control 220 oC for manifold, 240 oC for transfer line 20 µA 0.66 scane/sec 6 min 10 count 45 amu 45-450 amu 40 min None 25,000 µsec 20,000 count. Temperature Emission current Scan rate Filament delay Threshold Background mass Mass range Data aquisition time Ion preparation Maximum ionization time Target TIC. « ÜÂ " injector µ¶& oven Ë,1 Ø;'. ¥, ç4èD _*BÆ µd ¾ª. Table 1 U 2 ß 0'. Mass spectrometer1 ion trap ?S Varian Saturn. " :;r 2é 8 LLE µ¶ ±+ . 2200sD 0~650 Da ÚÐ r2| N>( electron. \(, µ§ GC/MS @ mass spectrum. impact ionization(EI) " ²=d'. MS. ê;'. Mass spectrum èD Ë. operation parameter ]7 Table 3& Z'.. fragmentation ±ë £T ìí(, fragmentation. à " \ 1 teflon á

(20) ]7 " methanolD âãd Ç fume hoodp ¶µ. ion @ ] ²=d Ç, ;r îïd ¾ª { rØ !T'. ²ð « ? d·, P·, ¸· U ¹º· I. "'.. VTBJ 8"d'.. . LLE d ¦§8= ¥j Ð@ " "À n-. . hexane&dichloromethane äÏ>, 5³·,, BÄ U XN Y \ uv wx1 = ] d {â

(21) 35å{ æ ß1 µ¶.

(22) ¾ª"(BHA: 4.4 ng/10 mL U BHT: 3.2 ng/10 mL)& ¾ª"(allethrin: 3.2 U fenvalerate:. Table 2. Temperature programming for antioxidant and insecticide. 2.9 ng/10 mL) FF 10 mLñ 2é c e 20 mL crimp seal cap vial ò(, "À 2 mL XNd '. Temp. (oC). Rate (oC/min). Hold (min). Total (min). − 08.0 10.0. teflon septaN óô capsD õöd Ç '. . 50 80 3000. 2.00 3.00 8.25. 2.00 8.75 40.00. BÄ7 3, 5, 10, 15, 20, 30, 60 2sD , 5³·,. Carrier gas: He (99.999%), Column flow: 1.2 ml/min(constant flow) 2003, Vol. 47, No. 6. 1 900, 500 U 0 rpmsD _*B÷'. øù B J"7 ú& "Àú ©¤ 2\1 102 Ç "Àú 200 µL é GCD injection'..

(23) 562.

(24) . Fig. 2. Comparison of TIC of n-hexane extraction with TIC of dichloromethane extraction (BHA: 0.44 ng/mL, BHT: 0.32 ng/mL, allethrin: 0.32 ng/mL, fenvalerate: 0.29 ng/mL).. Fig. 21 BHA, BHT, allethrin. \( fenvalerate Y. ^ peak1 m/z 57, 43p ß (, dichloromethane. ¥j Ð@ " "À n-hexane U dichloromethane. 2 r7 84( m/z 84, 49p ^ peakN ß. TIC Lûüý wx1 uv ^( 0'. N-. scan range 45~450 Da ÚÐp1 < 8sD n-hexane. hexane *[, þØ â

(25) 1 50,000~100,000 count. 3@

(26) 7 þØ â

(27) ßÉ '. y ê. ÚÐs , dichloromethane *[1 ÿ 400,000p. ; S/N3 dichloromethanesD d *[. ( 1,800,000 count /ÊN'.E7 dichloromethane. 1 13~27, n-hexanesD d *[1 32~46 ÚÐ. ý& · ý p h B. '. ±+p n-hexanesD 1 E 1.7~2.4

(28). Ä Ñb/( ±+ peak \

(29) < . 7 S/N 3 ßD wr2| \» . @ ßù <'. d, n-hexane7 2 r 86k. 0'. Fig. 37 BHA, BHT, allethrin U fenvalerate Y. Fig. 3. Influence of shaking (: not shaking, : shaking at 500 rpm, : shaking at 900 rpm) of sample on the extraction time profile (Y axis is detector response as peak area, X axis is adsorption time in minute). Journal of the Korean Chemical Society.

(30) Liquid Liquid Extraction-GC/MS (BHA, BHT) (Fenvalerate, Allethrin) . 563. Table 4. Recovery of each compound after liquid-liquid extraction. Solvent. n-hexane. dichloro -methane. n-hexane -NaCl(5 g). Compound BHA BHT Allethrin Fenvalerate BHA BHT Allethrin Fenvalerate BHA BHT Allethrin Fenvalerate. Recovery(%) 1 95.5 90.8 95.9 91.7 93.3 98.8 96.8 91.6 99.3 89.8 96.8 97.7. 2 89.7 94.3 91.7 88.4 89.8 91.9 96.3 92.8 92.8 90.1 92.9 98.2. 3 99.6 87.8 91.5 89.4 96.1 94.8 99.2 98.7 98.3 91.8 92.9 90.4. 4 95.2 89.5 90.7 93.3 88.2 88.8 90.8 89.8 97.8 92.0 95.8 94.6. 5 98.1 93.8 97.8 90.0 92.3 89.5 91.4 93.2 89.1 92.9 94.9 92.1. 6 91.4 96.4 93.2 92.1 93.6 99.6 92.8 95.4 89.9 96.8 97.5 90.2. 7 88.0 93.9 87.7 93.9 98.0 89.0 88.4 88.2 88.5 92.0 96.4 89.0. average 93.9 92.4 92.6 91.3 93.0 93.2. 93.7 92.8 93.7 92.2 95.3 93.2. Fig. 4. E.I. mass spectrum of fenvalerate.. n-hexanesD ¯ y, 5³·, U BÄ ±ë. ²=¯ 0sk, 3-phenoxy benzyl group d. uv ^( 0'. 900 rpmp 202 2¯. m/z 225, 181 peak, ßsD J] . EsD lF'. cd ÜÂ & þsD n-hexane,. ²= ;D n"¯ 0 E'. ÜÂ &D. dichloromethane U n-hexane-NaCl(5 g) "À ". ½ fenvalerate fragmentation ion] lT o. BHA, BHT, allethrin U fenvalerate ¦§ ê. ¯ 0sk, Fig. 51 N>d scheme . ;'. Å§ Table 4 ïÁ'. N-hexane U dichloromethane "Àp Å§7 FF 91.3~93.9% U 92.8~93.7%D t 1 Ý'. /¡ Fig. 2p ^ dichloromethane " d *[1 "À peak \

(31) < ßs k, þØ â

(32) noiseN ÊN'. d, n-hexaneNaCl(5 g)p1 92.2~95.3%Dp Å§ ÁÄ Ê NN 0'.. Fenvalerate mass spectrum7 Fig. 4 ßù Z m/z 125N base peak(, fenvalerate molecular weight peak= m/z 419 peak, B '. d, m/z 125 lT ç4= m/z 167& 152 peak 2003, Vol. 47, No. 6. Fig. 5. Scheme for fragmentation of fenvalerate..

(33)

(34) . 564. Fig. 6. E.I. mass spectrum of allethrin.. p ²='. d, 2-methyl-4-oxo-3-(2-propenyl)2-cyclopenten fragmentation d m/z 136 á , »Ó ²=¯ 0s , molecular weight peak= m/z 3037 {/ ~'. Fig. 77 allethrin fragmentation ion] N>d lT o ^ ( 0'. BHA1 molecular weight peak= m/z 180& [−CH3] N b lT m/z 165, [CO]N ( benzene (\N á Õb lT m/z 137 peakN ^ mass. Fig. 7. Scheme for fragmentation of allethrin.. spectrumsD ß(, BHT1 molecular weight peak = m/z 220 à [−CH3]N m/z 205 N ^ mass. ^( 0'. Allethrin mass spectrum7 Fig. 6p 1 Z. spectrumsD ²='. Ò 7 µN 358. 2,2-dimethyl-3-(2-methyl -1-propenyl) cyclopropane. Äç 'ë 3@ ç¿d fragmentation £T. fragmentation d m/z 123N base peakD,. E. ß'. Fig. 8 U 91 Ò d mass. sD½ l m/z 107& 91 peak mass spectrum. spectrum& fragmentation ion] N>d lT o. Fig. 8. E.I. mass spectrum of BHA (a) and BHT (b). Journal of the Korean Chemical Society.

(35) Liquid Liquid Extraction-GC/MS (BHA, BHT) (Fenvalerate, Allethrin) . 565. Fig. 9. Scheme for fragmentation of BHA and BHT.. FF ^( 0'. :d ÜÂ µ¶p BHA, BHT, allethrin U fenvalerate 4N/ V !V" TIC chromatogram ê; '. Fig. 10(a)1 4N/ V 2| "7 total ion count(TIC) chromatogram (, (b)1 TIC BHA £T ion= m/z 137& 180¡ "7 extracted ion chromatogram(XIC)'. #N/D Fig. 10(c)1 BHT £T ion= m/z 205 220, (d)1 allethrin £T ion= m/z 123,. \( (e)1 fenvalerate £. T ion= m/z 125 419¡ "7 XIC'. TIC 3@ XIC1 µP í²d 2| & ^ ( 0'.. Fig. 10. Chromatograms of BHA, BHT, allethrin, fenvalerate. (a) total ion count (TIC) chromatogram, (b) extracted ion chromatogram m/z 137+180 for BHA, (c) extracted ion chromatogram m/z 205+220 for BHT, (d) extracted ion chromatogram m/z 123 for allethrin and (e) extracted ion chromatogram m/z 125+419 for fenvalerate.. ng/10 mL) FF 10 mLñ é 20 mL crimp seal cap vial ò(, n-hexane 2 mL XNd ' teflon septa. . N óô capsD õö( 900 rpmp 202 Ö . {d4 ê; Ð@ ¾ªBJ 10 È| F. '. 102 Ö ;xBC Ç "Àú 200 µL é. F 5Å ñ ³$ ê; (, ] FF d æ& ¾ª D {d4(Limit. GCD injection'. µ% GC U MS µ¶ ±+ 2|d &D½. of Detection) '. Table 5 ïÁd Z,. !T {rØ7 Fig. 11 U 12p 1 Z. < ¾ª1(%RSD)1 1.41~5.34k, {d4. &ØT ß( 0'.. (LOD)1 0.071~0.159 ng/mL'. (BHA, BHT) U (fenvalerate, allethrin) ¥, ç4èD _* µd ¾ª"(3~18. ¸·(6'), ¹º·(5'), d·(6') U P·(3'). Table 5. Detection limit of BHA, BHT, allethrin and fenvalerate Measurement number 1. 2. 3. 4. 5. mean (ng/mL). SD (ng/mL). LOD* (ng/mL). 0.426 0.334 0.314 0.275. 0.434 0.352 0.414 0.280. 0.472 0.272 0.403 0.230. 0.505 0.293 0.303 0.240. 0.511 0.295 0.321 0.278. 0.470 0.309 0.351 0.261. 0.039 0.033 0.053 0.024. 0.118 0.098 0.159 0.071. Compounds Butylate hydroxy anisole Butylate hydroxy toluene Allethrin Fenvalerate. *LOD is computed as 3 times SD. 2003, Vol. 47, No. 6.

(36)

(37) . 566. Fig. 11. Calibration curves of BHA and BHT.. ;Îp d IBJ <sD BHA, BHT,. ? 5Å ³$ 2|d & Table 6 ïÁ. allethrin U fenvalerate 2| '.. c , 4 ·. '. VTBJ x ÜÂx Ä 1 +3.18~-. b(p, BHA, BHT, allethrin U fenvalerate1 {. 8.28% ÚÐ'. d, F ] < ¾ª(%RSD,. / ~'. « ? 8VT ½ ²= '. σ)1 6.39~13.4% ÚÐsk, 3σD½ {d. & Z VTBJ µ'. ), P· ;Î. 41 0.051~0.132 ng/mL '.. p d IBJ 1^: Ä ;x(. <Y. BHA, BHT, allethrin U fenvalerate FF 0.440, 0.320,. . . 0.320 U 0.290 ng/mL N'. Fig. 137 VT B J ¸·p éd BJ TIC chromatogram. Liquid-liquid extraction(LLE) d BHA, BHT, allethrin. ^( 0'. F BJD½ 37 minp b. U fenvalerate Y \ ÜÂp, 900 rpm 5³s. *d +,, ²=/ ~'. µd VTBJ «. D, 202 Ö n-hexane "À @ A 90% < Journal of the Korean Chemical Society.

(38) Liquid Liquid Extraction-GC/MS (BHA, BHT) (Fenvalerate, Allethrin) . 567. Fig. 12. Calibration curves of allethrin and fenvalerate.. '. § n-hexane& d "ÀD. 0.961D &ØT . ß( 0b BHA, BHT, allethrin. dichloromethane 0/¡ E7 ý& . U fenvalerate Y B2| N>'.. · ý hBÄ Ñbp peak \

(39). « LLE-GC/MS ¸·, ¹º·, d· U P·. <& noiseN ß'. Mass spectrum @ ²=. ;Îp d IBJ 8"BÆ « & BHA,. d &, F è fragmentation ion m/z7 '&. BHT, allethrin U fenvalerate1 b/ BJp, {. Z': BHA; 137, 165, 180, BHT; 205, 220, allethrin;. / ~'. BHA, BHT, allethrin U fenvalerate ¾. 91, 107, 123, 136, fenvalerate; 125, 152, 167, 181, 225,. ª" :;r P· IBJ N µd V. 419. F è {r-Ø7 linearity coefficientN 0.903~. TBJ « ? 8"BC &, x ÜÂx. 2003, Vol. 47, No. 6.

(40)

(41) . 568. Fig. 13. TIC chromatograms of the synthetic sample (upper) and a real sample at Nakdong treat. Table 6. Analysis of BHA, BHT, allethrin and fenvalerate in the synthetic sample Compound BHA BHT Allethrin Fenvalerate. Measurement Number 1. 2. 3. 4. 5. Mean ng/ml. 0.426 0.334 0.314 0.275. 0.434 0.352 0.403 0.280. 0.432 0.273 0.303 0.258. 0.505 0.295 0.293 0.241. 0.472 0.296 0.334 0.278. 0.454 0.310 0.329 0.266. Ä1 +3.18~ -8.28% ÚÐ N °l'. d, F ] < ¾ª(%RSD)1 6.39~13.4% (, 3σD½ {d4(LOD)1 0.051~0.132 ng/mL '. « 2|?7 0QT 0b 1sD '9d )* p 2|

(42) ¯ EsD

(43) '.. 1. Eichelberger, J. W.; Behymer, T. D.; Budde, W. L. Determination of organic compounds in drinking water by LSE and capillary column GC/MS, Revision 2.2EPA EMSL-Ci, May 1991. 2. Pang, G. F.; Fan, C. L.; Chao, Y. Z.; Zhao, T. S. J. Chromatogr. 1994, 667, 348. 3. Pang, G. F.; Fan, C. L.; Chao, Y. Z.; Zhao, T. S. J. AOAC Inter. 1994, 77, 738. 4. Munch, J. W. Determination of organic compounds in drinking water by liquid solid extraction and capillary column GC/MS, National Exposure Research Labora-. SD ng/ml. RSD %. Theor. ng/ml. Exper. ng/ml. Devia. %. 0.034 0.032 0.044 0.017. 7.49 10.3 13.4 6.39. 0.440 0.320 0.320 0.290. 0.454 0.310 0.329 0.266. +3.18 −3.13 +2.81 -8.28. tory Office of Research and Development U.S. Environmental Protection Agency, 1995. 5. Tena, M. T.; Valcarcel, M.; Hidalgo, P. J.; Ubera, J. L. Anal. Chem. 1997, 69, 521. 6. Ramesh, A.; Balasubramanian, M. Analyst, 1998, 123, 1799. 7. Eriksson, M.; Faldt, J.; Dalhammar, G.; Borg-Karlson, A. K. Chemospher. 2001, 44, 1641. 8. Brito, N. M.; Navickiene, S.; Polese, L.; Jardim, E. F. G.; Abakerli, R. B.; Ribero, M. L. J. Chromatogr. A, 2002, 957, 201. 9. Brossa, L.; Marcé, R. M.; Borrull, F.; Pocurull, E. J. Chromatogra. A. 2002, 963, 287. 10. Petrovic, M.; Eljarrat, E.; de Alda, M. J. L.; Barcelo, D. J. Chromatogra. A. 2002, 974, 23. 11. Kambia, K.; Dine, T.; Gressier, B.; Germe, A.-F.; Luyckx, M.; Burnet, C.; Michaud, L.; Gottrand, F. Biomed. Sci. Appl. 2001, 755, 297. 12. Ramesh, A.; Balasubramanian, M. Analyst 1998, 123, 1799. 13. Jara, S.; Lysebo, C.; Greibrokk, T.; Lundanes, E. Analy. Chimica Acta 2000, 407, 165.. Journal of the Korean Chemical Society.

(44)