Mahato Ansuman, Vyas Samir, Chatterjee Niladri Sekhar
Agilent Technologies India Pvt Ltd, IMT Manesar Sector 8, Plot CP11, Gurugram 122051, Haryana, India.
Indian Council of Agricultural Research, Central Institute of Fisheries Technology, CIFT Junction, Matsyapuri P O, Willingdon Island, Cochin 682029, Kerala, India.
J AOAC Int. 2020 Jan 1;103(1):73-77. doi: 10.5740/jaoacint.19-0207.
In India, fortification of cereals with folic acid has been voluntary for many years. However, The Food Safety and Standards Authority of India's recent Fortification of Foods Regulations (2018) has prompted us to develop and validate a simple analytical method for estimation of folic acid in fortified cereals.
The aim was to develop and validate a simple and rugged HPLC-UV method for quantitative analysis of folic acid in fortified rice and wheat flour.
The enzymatic sample extract was diluted with phosphate buffer, centrifuged, filtered, and then passed slowly through an immunoaffinity cartridge for cleanup. Folic acid in the sample extract was retained by the cartridge and subsequently eluted with 30% acetonitrile [+0.2% trifluoroacetic acid (TFA)]. The elute was collected and analyzed by HPLC-UV at 280 nm. The chromatographic separation of folic acid was achieved on an Agilent Poroshell SB-C18 column (3.0 × 100 mm, 2.7 µm) with 0.1% TFA in methanol as mobile phase.
The linearity range of the vitamin was established in the concentration range of 50-800 µg/L, and the regression coefficient was more than 0.999. The LOQ was 5 µg/L. The average spike recovery values of folic acid in rice and wheat flour samples were 90.9 and 80.5%, respectively. The method was subjected to an interlaboratory validation; eight accredited food testing laboratories across India participated in it and resulted in satisfactory z-scores for the reported results.
The method will be useful in regulatory compliance testing of folic acid in fortified cereals and processed products.
A sensitive analysis method is reported for estimation of folic acid in fortified rice and wheat flour. The scope, selectivity, repeatability, and reproducibility of the method establishes it as fit for regulatory compliance check purposes.
在印度，谷类食品强化叶酸已实行多年，但一直是自愿行为。然而，印度食品安全标准管理局最近出台的《食品强化法规》（2018年）促使我们开发并验证一种用于测定强化谷类食品中叶酸的简单分析方法。
旨在开发并验证一种简单且耐用的高效液相色谱 - 紫外检测法，用于定量分析强化大米和小麦粉中的叶酸。
酶解后的样品提取物用磷酸盐缓冲液稀释，离心、过滤，然后缓慢通过免疫亲和柱进行净化。样品提取物中的叶酸被亲和柱保留，随后用30%乙腈[+0.2%三氟乙酸（TFA）]洗脱。收集洗脱液，在280nm波长下用高效液相色谱 - 紫外检测法进行分析。叶酸的色谱分离在安捷伦Poroshell SB - C18柱（3.0×100mm，2.7μm）上进行，流动相为含0.1%TFA的甲醇。
该维生素的线性范围在50 - 800μg/L浓度区间内建立，回归系数大于0.999。定量限为5μg/L。大米和小麦粉样品中叶酸的平均加标回收率分别为90.9%和80.5%。该方法进行了实验室间验证；印度全国八家获得认可的食品检测实验室参与其中，报告结果的z值令人满意。
该方法将有助于强化谷类食品和加工产品中叶酸的法规合规性检测。
报道了一种用于测定强化大米和小麦粉中叶酸的灵敏分析方法。该方法的范围、选择性、重复性和再现性表明其适用于法规合规性检查目的。
