Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Optimization of HPLC Method and Clean-up Process for Simultaneous and Systematic Analysis of Synthetic Color Additives in Foods

식품 중 타르색소의 동시분석 및 계통분석을 위한 HPLC 분석조건 및 정제과정 확립

  • Park, Sung-Kwan (Division of Food Additives Evaluation, Korea Food and Drug Administration) ;
  • Hong, Yeun (Division of Food Additives Evaluation, Korea Food and Drug Administration) ;
  • Jung, Yong-Hyun (Division of Food Additives Evaluation, Korea Food and Drug Administration) ;
  • Lee, Chang-Hee (Division of Food Additives Evaluation, Korea Food and Drug Administration) ;
  • Yoon, Hae-Jung (Division of Food Additives Evaluation, Korea Food and Drug Administration) ;
  • Kim, So-Hee (Division of Food Additives Evaluation, Korea Food and Drug Administration) ;
  • Lee, Jong-Ok (Division of Food Additives Evaluation, Korea Food and Drug Administration)
  • 박성관 (식품의약품안전청 식품첨가물평가부) ;
  • 홍연 (식품의약품안전청 식품첨가물평가부) ;
  • 정용현 (식품의약품안전청 식품첨가물평가부) ;
  • 이창희 (식품의약품안전청 식품첨가물평가부) ;
  • 윤혜정 (식품의약품안전청 식품첨가물평가부) ;
  • 김소희 (식품의약품안전청 식품첨가물평가부) ;
  • 이종옥 (식품의약품안전청 식품첨가물평가부)
  • Published : 2001.02.28
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Abstract

To develop a method for separation process using Sep-pak $C_18$, simultaneous and systematic analysis of 8 permitted and 11 non-permitted synthetic food colors in Korea, optimization of analysis conditions for reverse phase ion-pair high performance liquid chromatography was carried out. For the best result of Sep-pak $C_18$ separation the pH of color standard mixture solution was $5{\sim}6$ and 0.1% HCl-methanol solution were set as eluent. The colors eluated from Sep-pak $C_18$ cartridge were determined and confirmed by high performance liquid chromatography with a photodiode array detector at 420 nm for yellow colors type, at 520 nm for red colors type, at 600 nm for blue and green colors type and at 254 nm for mixed colors. Conditions for HPLC analysis were as follows: column, Symmetry $C_18$ (5 m, 3.9 mm $i.d.{\times}150\;mm$); mobile phase, 0.025 M ammonium acetate (containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (65 : 25 : 10) and 0.025 M ammonium acetate(containing 0.01 M tetrabutylammonium bromide) : acetonitrile : methanol (40 : 50 : 10); flow rate, 1 mL/min. It takes 35 minutes for simultaneaus analysis and 18 minutes for systematic analysis. The detection limits range of each colors were $0.01{\sim}0.05\;{\mu}g/g$.

이온쌍고속액체크로마토그래피를 이용하여 현재 우리나라에서 식품에 사용이 허용된 타르색소 8종과 일본등 외국에서는 허용되어 있으나 우리나라에서 허용되어 있지 않은 11종 등 총 19종의 동시 및 계통분석법을 확립하였다. HPLC에 사용한 컬럼은 Symmetry $C_18$, 이동상은 0.01 M TBA-Br이 함유된 0.025 M 초산암모늄용액-아세토니트릴-메탄올(65 : 25 : 10)과 0.01 M TBA-Br이 함유된 0.025 M 초산암모늄용액-아세토니트릴-메탄올(40 : 50 : 10)를 구배용매조성법(gradient mode)으로 사용하였다. 자외선 검출기의 파장은 동시분석의 경우 254 nm, 계통분석은 황색계통 420 nm, 적색계통 520 nm 그리고 청색 및 녹색계통은 620 nm로 설정하였다. 이때 색소들의 검출한계는 적색계통이 $0.05\;{\mu}g/g$, 황색계통은 $0.03\;{\mu}g/g$, 청색 및 녹색계통은 $0.01\;{\mu}g/g$이었다. Sep-pak $C_18$을 이용한 색소의 정제 방법은 혼합색소표준용액의 pH를 $5{\sim}6$으로 조정하고 0.1% TBA-Br을 가하여 색소를 보유시킨 다음 0.1% 염산-메탄올로 색소를 용출하여 HPLC로 분석하였다. 19종의 타르색소를 동시분석하는데 35분이, 계통분석시에는 18분 정도가 소요되었고 두 경우 모두 양호한 분리를 보였다.

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