Identification and Determination of the Isomer and Subsidiary Color in Food Blue No. 2 (Indigo Carmine) by LC/MS and HPLC

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  • TSUJI Sumiko
    Division of Food Chemistry, Osaka Branch, National Institute of Health Sciences National Institute of Health Sciences
  • NAKANO Maki
    Division of Food Chemistry, Osaka Branch, National Institute of Health Sciences Saito Lab., AnGes MG, Inc.
  • FURUKAWA Mizuki
    Division of Food Chemistry, Osaka Branch, National Institute of Health Sciences Tokyo Center, Japan Inspection Association of Food and Food Industry Environment
  • YOSHII Kimihiko
    Division of Food Chemistry, Osaka Branch, National Institute of Health Sciences Pharmaceutical Affair Division, Department of Public Health and Welfare, Osaka Prefectural Government
  • TONOGAI Yasuhide
    Division of Food Chemistry, Osaka Branch, National Institute of Health Sciences Test Inspection Center, Osaka Pharmaceutical

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  • ＬＣ／ＭＳおよびＨＰＬＣによる食用青色２号（インジゴカルミン）中の異性体および副成色素の同定・定量
  • LC MS オヨビ HPLC ニ ヨル ショクヨウ アオイロ 2ゴウ インジゴカルミン チュウ ノ イセイタイ オヨビ フクセイシキソ ノ ドウテイ テイリョウ

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Abstract

CFR and JECFA specify that the total color in FD&C Blue No. 2 (B2; Indigo Carmine, Indigotine, Food Blue No. 2) is not less than 85%, its isomer (B2iso) in B2 is not more than 18%, and its subsidiary color (B2sub) in B2 is not more than 2% (CFR) or 1% (JECFA). Japan's Specifications and Standards for Food Additives, 7th Edition, specifies that the total color in B2 is not less than 85.0% and other color materials in B2 are not detected by paper chromatography. LC/MS and HPLC were employed to identify and determine the main component (B2m) of B2, B2iso, and B2sub. The pseudo molecular ions (B2m and B2iso: [M-2Na+H]-, m/z=421; B2sub: [M-Na]-, m/z=341) of each color material were obtained and identified by LC/MS based on their absorptions and mass spectra. The contents of B2iso and B2sub in B2 samples (certified samples from fiscal year 1998 to fiscal year 2002) were determined by HPLC using calibration curves for the standards of B2m and B2iso. The contents of B2iso in most samples were less than 10%, and the contents of B2sub in all samples were not more than 1%. All of them were within the regulatory limits set by the CFR and JECFA.

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