Determination of fluorescein and brominated fluoresceins in the color additive D&C Orange No. 5 and its lakes using high-performance liquid chromatography. Issue 12 (2nd December 2019)

• Record Type: Journal Article
• Authors: James, India C.; Lazo-Portugal, Rodrigo; Perez-Gonzalez, Marianita; Weisz, Adrian
• Is Part Of: Food additives & contaminants. Volume 36:Issue 12(2019)
• Page Start: 1800
• Page End: 1807
• Publication Date: 2019-12-02
• Subjects: Color additive D&C Orange No. 5 -- monobromofluorescein -- dibromofluorescein -- tribromofluorescein -- tetrabromofluorescein -- HPLC; Food additives -- Periodicals; Food contamination -- Periodicals; 664.06
• DOI: 10.1080/19440049.2019.1664773
• Languages: English
• ISSNs: 1944-0049

Abstract

ABSTRACT: The colour additives D&C Orange No. 5 (O5) and its lakes (O5L) are subject to batch certification by the U.S. Food and Drug Administration (FDA) to ensure compliance with specifications in the Code of Federal Regulations (CFR). The present study reports the development of a high-performance liquid chromatography (HPLC) method for the quantitative determination of seven CFR-specified components in O5 and O5L – fluorescein and six brominated fluoresceins. The analytes were quantified using six-point calibration curves with data points (w/w) that ranged as follows: 20.0–70.0% for 4ʹ, 5ʹ-dibromofluorescein; 9.8–44.1% for 2ʹ, 4ʹ, 5ʹ-tribromofluorescein; 1.01–15.2% for 2ʹ, 4ʹ, 5ʹ, 7ʹ-tetrabromofluorescein; 0.10–3.12% for 2', 4ʹ-dibromofluorescein; 0.10–3.06% for 2ʹ, 5ʹ-dibromofluorescein; 0.11–2.85% for 4ʹ-bromofluorescein; and 0.10–2.02% for fluorescein. For all seven analytes, the HPLC instrument response was linear (R 2 > 0.999) over the tested concentration ranges and the limits of detection (0.01–1.55%) were well below the CFR-specified levels. Other validation data showed good analyte recovery (87.91–101.73%) as well as method precision measured by the relative standard deviation (0.53–1.56%). The new method was applied to the analysis of test portions from 15 batches of O5 and eight batches of O5L submitted to FDA for certification by domestic and foreign manufacturers. Compared to the thin-layer chromatography/spectrophotometric procedure currently used for routine batch-certification analyses, the new method was found to be more sensitive, simpler to implement, and significantly faster, requiring 25 minutes rather than six hours to analyse one sample.