Belai Nebebech, Weisz Adrian
U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, Office of Cosmetics and Colors, 5001 Campus Dr, College Park, MD 20740.
J AOAC Int. 2019 May 1;102(3):936-941. doi: 10.5740/jaoacint.18-0260. Epub 2018 Nov 16.
Certain impurities in the color additives drug and cosmetic (D&C) Red No. 21 (R21), D&C Red No. 22 (R22), and their lakes are limited to levels specified in the Code of Federal Regulations (CFR) and are quantified by the U.S. Food and Drug Administration in batches of these color additives submitted for certification. Currently, a lengthy and tedious method based on gravity flow elution column chromatography is used to quantify the following CFR-specified impurities: the intermediate, phthalic acid (PhthAc); the manufacturing by-products, 2-(3',5'-dibromo-2',4'-dihydroxybenzoyl)benzoic acid (Br2BBA); and brominated resorcinol. "Brominated resorcinol" implies the sum of all possible brominated resorcinols, but the current work focused on 2,4,6-tribromoresorcinol (Br3R) as the most probable side-reaction product. An improved method was needed to quantify PhthAc, Br2BBA, and Br3R in R21, R22, and their lakes. A rapid ultra-HPLC (UHPLC) method was developed to replace the gravity flow method for quantitative determination of PhthAc, Br2BBA, and Br3R. PhthAc, Br2BBA, and Br3R were quantified by using five-point calibration curves with data point ranges of 0.11-1.55, 0.06-0.77, and 0.04-0.61% by weight, respectively. LODs for the analytes ranged from 0.01 to 0.03%. Recoveries of the analytes ranged from 90.6 to 99.9%. The UHPLC method is accurate and significantly more rapid than the gravity -flow method, requiring approximately 7 min as compared with 6 h to detect PhthAc, Br2BBA, and Br3R in one sample. A rapid UHPLC method was developed to determine CFR-specified impurities in the color additives D&C Red Nos. 21 and 22 and their lakes.
色素添加剂药品及化妆品（D&C）红21号（R21）、D&C红22号（R22）及其色淀中的某些杂质含量须符合美国联邦法规（CFR）的规定，并由美国食品药品监督管理局对提交认证的这些色素添加剂批次进行定量检测。目前，采用基于重力流洗脱柱色谱法的冗长繁琐方法来定量检测以下CFR规定的杂质：中间体邻苯二甲酸（PhthAc）、生产副产物2-(3',5'-二溴-2',4'-二羟基苯甲酰基)苯甲酸（Br2BBA）以及溴代间苯二酚。“溴代间苯二酚”指所有可能的溴代间苯二酚的总和，但当前工作聚焦于最可能的副反应产物2,4,6-三溴代间苯二酚（Br3R）。需要一种改进方法来定量检测R21、R22及其色淀中的PhthAc、Br2BBA和Br3R。开发了一种快速超高效液相色谱（UHPLC）方法来替代重力流方法，用于定量测定PhthAc、Br2BBA和Br3R。通过使用五点校准曲线对PhthAc、Br2BBA和Br3R进行定量，数据点范围分别为0.11 - 1.55%、0.06 - 0.77%和0.04 - 0.61%（重量）。分析物的检测限范围为0.01%至0.03%。分析物的回收率范围为90.6%至99.9%。UHPLC方法准确且比重力流方法显著更快，检测一个样品中的PhthAc、Br2BBA和Br3R大约需要7分钟，而重力流方法需要6小时。开发了一种快速UHPLC方法来测定色素添加剂D&C红21号和22号及其色淀中CFR规定的杂质。
