In Japan, twelve color additives, namely, Amaranth (R2), Erythrosine (R3), Allura Red AC (R40), Ponceau 4R (R102), Phloxine (R104), Rose Bengal (R105), Acid Red (R106), Tartrazine (Y4), Sunset Yellow FCF (Y5), Fast Green FCF (G3), Brilliant Blue FCF (B1), and Indigo Carmine (B2) are permitted for use in food and regulated by the Food Sanitation Act. Identification tests, purity tests, and assays are defined to confirm their quality and safety in the Japan’s Specifications and Standards for Food Additives (JSSFA). In the 9th JSSFA, the maximum limits for impurities (unreacted raw materials and products of side reactions) and subsidiary colors in these color additives will be defined to promote international harmonization. Therefore, we developed methods of determining impurities and subsidiary colors in the color additives using high-performance liquid chromatography (HPLC).<br/>A gradient of two mobile phases (ammonium acetate solution and acetonitrile aqueous solution), a wavelength of 254 nm to detect impurities, and a visible wavelength to detect subsidiary colors were used. Standard curves were used to determine impurities; the area% ((total peak area − main color peak area)/total peak area × color content) was used to determine subsidiary colors.<br/>In the impurity recovery test from R3, G3, B1, and B2, the developed method exhibited satisfactory recoveries. The amounts of impurities and/or subsidiary colors were less than their maximum limits as determined from the impurity and subsidiary color survey for R2, R3, R102, Y4, G3, B1, and B2 commercial products. Therefore, it can be noted that the HPLC method is reliable and can be implemented for routine analysis of these color additives. The maximum limits for impurities and subsidiary colors in R104, R105, and R106 will be defined as 6%, 4.5%, and 10%, respectively, in the 9th JSSFA based on the survey of their commercial products.

Development of a specification method to determine unreacted raw materials, products of side reactions, and subsidiary colors in color additives using high-performance liquid chromatography

254th National Meeting (2017)

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