350 De Kruijf 4) MPEPPP BP 4 Maeda 5) MPPPiPP BP ibp 5 HPLC HPCE Shu Ping 6) HPCE SDS Capillary Zone Electrophoresis CZE MPEPPP BP 10 CZE 139 BHA BHT

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Vol. 12 No. 2 2002 349 355 Λ1 Λ2 Λ1 MEKC MPEP PPiPPBPiBP BeP BHA BHT 9 20 mm SDS 30 mm ph 9.35 15 kv 11 5 80 mg L 1 0.99 R.S.D.(%) 0.33 BHT R.S.D. 10 %mg L 1 MP 2.28EP 3.28PP 2.28iPP 3.62BP 3.62iBP 1.54BeP 6.46BHA 9.17BHT 6.17 MEKC 3 1 5 BHT 13 4 EP PP ipp BP ibp 5 5 MP BeP Na MP 3 13 158 MPEPPP BP 1;2) Fujii 3) 5 EPPP ippbpibp HPCE SDS Micellar Electrokinetic Capillary Electrophoresis MEKC BP HPLC Λ1 Λ2 701-0193 288 349

350 De Kruijf 4) MPEPPP BP 4 Maeda 5) MPPPiPP BP ibp 5 HPLC HPCE Shu Ping 6) HPCE SDS Capillary Zone Electrophoresis CZE MPEPPP BP 10 CZE 139 BHA BHT BHA BHT 7) BHA 8) BHA BHT BHA BHT BHABHTMEKC BHT 1 Na 2 B 4 O 7 10H 2 O MW=381.37- C 6 H 10 O 5 7 MW=1135.00- CDSDSCH 3 (CH 2 ) 11 OSO 3 NaMW=288.38 BHA 2-and3-Tert-Butyl- 4 - HydroxyanisoleC 11 H 16 O 2 MW=180.25 BHT 3 5 -Di-Tert-Butyl-HydroxytolueneC 15 H 24 O MW=220.35HO 3 C 6 H 2 COOCH 2 CH 2 CH 3 MW=212.20 PG HPLC C 2 H 5 OH MW=46.07CH 3 CN MW=41.05HPLC CH 3 OH MW=32.04HPLC MPHOC 6 H 4 COOCH 3 MW=152.15EP HOC 6 H 4 COOC 2 H 5 MW=166.17PPHOC 6 H 4 COOCH 2 CH 2 CH 3 MW=180.20iPPHO C 6 H 4 COOCH(CH 3 ) 2 MW=180.20BPHOC 6 H 4 COOCH 2 CH(CH 3 ) 2 MW=194.23 ibp HOC 6 H 4 COOCH 2 CH(CH 3 ) 2 MW=194.23 BePHOC 6 H 4 COOC H 2 C 6 H 5 MW=228.25 2 7 MPEPPP ippbpibp BeP 10.0mg 50%10mL BHA BHT 10.0mg A A 10 ml 50% DISMIC 25 0.20 m 10 0.763g20mM 100mL ph 9.35 SDS 0.865g30mM 30 3 512ch CAPI 3001 75 m I.D.500 mm L. 15 kv25 200nm10 nl15 mm30 sec 4 7 BHABHT 5 80 mg L 1 5 LOD 3 Repeatability 7 BHA BHT50 mg L 1 9 Relative Standard

MEKC BHABHT 351 Deviation R.S.D. 5 3 A BC AP BJ CS A B 20mL 1 ml 1mL 1 50% 8mL C 20mL 1g 3mL 1 50% 6mL3,000 r.p.m.10 10 1 PG 7 BHA BHT 9 20 mg L 1 1 1 BeP BHA 7 BHA BHT 11 SDS 20253050mM 30mM SDS 20mM BHA BHT BHA BHT 15 mm -CD 25 mm SDS 20mM MEKC 9) BHA BHT 5.59.3 - CD 10) -CD 2 5 80 mg L 1 1 BHABHT 2 LOD mg L 1 pg=10 12 g Shu Ping 4) CZE ng MP 0.97EP 1.09PP 1.22BP 1.33 1 Electropherogram and UV spectra of the mixed standard solution of parabens and antioxidantspeaks1 propyl gallate (internal standard)2 methyl paraben (MP)3 ethyl paraben (EP)4 isopropyl paraben (ipp)5 propyl paraben (PP)6 isobutyl paraben (ibp) 7 butyl paraben (BP) 8 benzyl paraben (BeP) 9 butylated hydroxyanisole (BHA) 10butylated hydroxytoluene (BHT)

352 LOD Shu Ping 1/30 1 /50 1 2 Calibration graphs of parabens and antioxidants by MEKC Limits of detection of parabens and antioxidants by MEKC. C MP BP 4 BHT 100 g 1.0 g Shu-Ping 4) 11 CZE 4 MP MP MP MP 3 Repeatability formekc. 3 3 R.S.D.% 0.190.33 BHT 10 % BHA BHA 4 Concentrations of parabens and BHT in three cosmetic products 4 BHT 2 A A BHT 2 1 MP BHT 3 B B BHT 5 MPEP PPBPiBP BHT 4 C 12332 BHT 3 1 BHT BHT BHT

MEKC BHABHT 353 2 Electropherogram and an UV spectrum of methyl paraben (Peak 1 ) in sample A 3 Electropherogram and UV spectra of five parabens and BHT in sample B Peaks 1 MP 2,EP 3 PP 4 ibp 5 BP 6 BHTSee the legend of Fig. 1 for abbreviations 4 Electropherogram and UV spectra of methylparaben and butylparaben in sample C. Peaks 1 MP 2 BPSee the legend of Fig. 1 for abbreviations

354 1Routledge EJParker JOdum JAshdy J and Sumpter JP Some Alkyl Hydroxy Benzoate Preservatives (Parabens) are EstrogenicToxicology and Applied Pharmacology 15312 191998 2Hossaini ALarsen JJ and JC Lack of Oestrogenic Effects of Food Preservatives (Parabens) in Uterotrophic AssaysFood and Chemical Toxicology 38319 3232000 3Fujii TKawabe S Simultaneous Determination of Sodium Benzoate and p Hydroxybenzoate Esters Using High performance Capillary ElectrophoresisKawasaki Journal of Medical Welfare7(1)33 412001 4N De KruijfA SchoutenM AHRijkL A Pranoto Soetardhi Determination of preservatives in cosmetic products IIHigh performance liquid chromatographic identificationjournal of Chromatography 469317 3281989 5Maeda YYamamoto MOwada KSato SMasui T High prerformance liquid chromatographic determination of six p hydroxybenzoic acid esters in cosmetics using sep pak florisil cartridges for sample pre treatmentjournal of Chromatography 410413 4181987 6Shu Ping WChing Li C Determination of parabens in cosmetic products by supercritical fluid extraction and capillary zone electrophoresisanalytica Chimica Acta 37785 931998 7Ito NHagiwara AShibata MOgiso T and Fukushima S Induction of squamous cell carcinoma in the forestomach of F344 rats treated with butylated hydroxyanisolegann 732332 334, 1982 8Bao TZJoseph LRobert TR and Allan HC Effect of Dietary 2(3) tert Butyl 4 hydroxyanisole on the Metabolism and Action of Estradiol and Estrone in Female CD 1 MiceCancer Research 572419 2427, 1997 9 : 111133 139, 2001 10 : 12, 2000 141031

MEKC BHABHT 355 Simultaneous Determination of Parabens, Butylated Hydroxyanisole and Butylated Hydoroxytoluene in Cosmetic Products Using Micellar Electrokinetic Capillary Electrophoresis Satoko KAWABE, Rie YANAI and Toshiko FUJII (Accepted Oct. 31, 2002) Key words : micellar electorokinetic capillary electrophoresis (mekc), parabens, butylated hydroxyanisole, butylated hydroxytoluene, cosmetics. Abstract A rapid and simple micellar electrokinetic capillary electrophoresis method was developed for the simultaneous determination of the parabens, methyl-, ethyl-, propyl-, isopropyl-, butyl-, isobutyl- and benzyl p-hydroxybenzoate and two phenolic antioxidants, butylated hydroxyanisole and butylated hydroxytoluene, which were widely used in cosmetic products. The nine chemicals were successfully separated within 11 minutes using a 20 mm tetraborate buffer containing 30 mm sodium dodecyl sulfate (ph 9.35) with a constant voltage of 15 kv. The correlation coefficients of the linear calibration graphs for these chemicals exceeded 0.99 in concentrations ranging between five and 80 mg L 1. The repeatability of the migration times of the chemicals was satisfactory [R.S.D.(%) 0.33], under optimum separation conditions, and peak areas and heights had R.S.D. values less than 10 %. The detection limits (mg L 1 )were 2.28 for MP, 3.28 for EP, 2.28 for PP, 3.62 for ipp, 3.62 for BP, 1.54 for ibp, 6.46 for BeP, 9.17 for BHA and 6.17 for BHT. Using the present method, five kinds of parabens and butylated hydroxytoluene were detected in one of three commercial cosmetics. Correspondence to : Satoko KAWABE Department of Clinical Nutrition, Faculty of Medical Professions, Kawasaki University of Medical Welfare Kurashiki, 701-0193, Japan (Kawasaki Medical Welfare Journal Vol.12, No.2, 2002 349 355)