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J. L. Belsky, A. J. Ashley, P. A. Bhatt, K. V. Gilbert, H. R. Joyce, C. Pan, H. Pappa and S. Z. Wahab, “Optimization of the Water-Insoluble Procedures for USP General Chapter Residual Solvents<467>,” AAPS PharmSciTech, Vol. 2, No. 2, 2010, pp. 994-1004. doi:10.1208/s12249-010-9460-6

has been cited by the following article:

  • TITLE: Analysis of Residual Solvents in Annatto Extracts Using a Static Headspace Gas Chromatography Method

    AUTHORS: Yusai Ito, Kyoko Ishizuki, Wakana Sekiguchi, Atsuko Tada, Takumi Akiyama, Kyoko Sato, Takeshi Yamazaki, Hiroshi Akiyama

    KEYWORDS: Annatto Extracts; Bixin; Norbixin; Headspace Gas Chromatography; Residual Solvents

    JOURNAL NAME: American Journal of Analytical Chemistry, Vol.3 No.9, September 28, 2012

    ABSTRACT: An analytical method for the quantification of residual solvents in annatto extracts, natural food colorants, was established using a static headspace gas chromatography (HSGC) coupled with a flame ionization detector (FID). As a sample diluent in a headspace sampling, dimethylformamide (DMF) was selected owing to its high capacity for dissolving both bixin-based and norbixin-based annatto extracts. The quantification of residual solvents was performed using the external standard method. The linearity of the calibration curves was assured with relative coefficients (R2) that were greater than 0.999. The recoveries of all standard solvents spiked in the annatto extracts were in the range from 95.1% to 107.1% to verify the accuracy and the relative standard deviation (RSD%) values (n = 3) were in the range from 0.57% to 3.31%. The quantification limits (QL) were sufficiently lower than the limits specified by Joint FAO/WHO Expert Committee on Food Additives (JECFA). With the established HSGC method, six residual solvents (methanol, ethanol, 2-propanol, acetone, ethyl acetate, and hexane) in 23 commercial annatto-extract products that consist of seven bixin-based and 16 norbixin-based products were quantified. The levels of residual ethyl acetate and hexane in all products were lower than the specified limits of JECFA. However, three samples of bixin-based products showed higher levels of residual 2-propanol (approximately 313.9 - 427.7 ppm) than the specified limit. Other bixin products also showed higher concentrations of residual methanol (approximately 166.6 - 394.7 ppm) and residual acetone (approximately 75.2 - 179.8 ppm) than the limits of JECFA. In the case of norbixin-based products, nine samples showed higher levels of residual acetone (approximately 42.6 - 139.5 ppm) than the limits of JECFA. This is the first survey of residual solvents in annatto extracts using the validated HSGC method.