Envirocare Labs Pvt. Ltd, A-7, Enviro House, Wagle Industrial Estate, Thane, 400604, India.
Syngene International Ltd, Syngene Regional Laboratory, ANRD, BSEZ Unit-VI, Block S-5, Plot No. 2 & 5, Phase-IV, Bomasandra Jigani Link Rd Bangalore 560100 India.
J AOAC Int. 2020 Jan 1;103(1):3-8. doi: 10.5740/jaoacint.19-0199.
Ensuring the quality of infant and pediatric formulas and adult nutritionals is of utmost importance for the health and safety of rapidly urbanizing Indian population. B12 is an important water-soluble vitamin, which is fortified externally in such nutritional formulations. The Bureau of Indian Standards (BIS) has a recommended microbiological assay-based method for determination of vitamin B12 that is not precise and accurate enough to meet the label claim requirements of infant, adult, and/or pediatric nutritionals. The AOAC Official Method 2011.10 was originally developed under the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) for the determination of vitamin B12 in infant and pediatric formulas and adult nutritionals. However, those SPIFAN matrixes did not contain malt and other indigenous cereal and legume flour (with or without cocoa powder), which are commonly found in Indian formulations. Thus, there is a need to replace this method with a more precise and accurate method.
This study was undertaken to validate the AOAC Official Method 2011.10 on vitamin B12 in 'Indian' infant and pediatric formulas and adult nutritionals.
The single-laboratory validation (SLV) of AOAC Method 2011.10 was carried out as per the AOAC Guidelines in six Indian pediatric and adult nutritional formulas to verify its fitness for purpose. Cobalamin in the sample was converted to cyanocobalamin on treatment with potassium cyanide. The sample was then subjected to clean up through a C18 cartridge. Vitamin B12 in the eluted extract was separated from other components using size-exclusion column chromatography followed by a C18 column. The HPLC analysis was carried out at 550 nm.
Diastase treatment and C18 solid-phase extraction cleanup satisfactorily removed the matrix interference. The relative standard deviation of the determined values in 30 samples each from 6 selected Indian products and NIST SRM 1849a was <20%. The average recoveries for the spiked recovery samples ranged from 91.75 to 101.14%.
Method 2011.10 met the standard method performance requirements set forth by the AOAC SPIFAN. Therefore, we recommend the Method 2011.10 for adoption as the BIS official method for the analysis of vitamin B12 in 'Indian' infant and pediatric formulas and adult nutritionals.
This was the first SLV project that the AOAC India section undertook to extend the scope of the AOAC Method 2011.10 for vitamin B12 analysis by validating it in 'Indian' infant and pediatric formulas and adult nutritionals.
确保婴儿配方奶粉和成人营养品的质量对于迅速城市化的印度人口的健康和安全至关重要。B12 是一种重要的水溶性维生素，在这些营养配方中外部强化添加。印度标准局（BIS）有一个基于推荐微生物检测的方法来测定维生素 B12，但该方法不够精确和准确，无法满足婴儿、成人和/或儿科营养品的标签声称要求。AOAC 官方方法 2011.10 最初是由 AOAC 婴儿配方和成人营养品利益相关者小组（SPIFAN）制定的，用于测定婴儿配方和成人营养品中的维生素 B12。然而，这些 SPIFAN 基质中不包含麦芽和其他本土谷物和豆类面粉（含或不含可可粉），这些在印度配方中很常见。因此，需要用更精确和准确的方法来替代它。
本研究旨在验证 AOAC 官方方法 2011.10 在“印度”婴儿配方和儿科配方及成人营养品中的维生素 B12 的适用性。
根据 AOAC 指南，在六份印度儿科和成人营养配方中进行了单实验室验证（SLV），以验证其适用性。用氰化钾处理将样本中的钴胺素转化为氰钴胺素。然后，通过 C18 筒进行样品净化。用尺寸排阻柱色谱法和 C18 柱将洗脱提取物中的维生素 B12 与其他成分分离。在 550nm 处进行 HPLC 分析。
淀粉酶处理和 C18 固相萃取净化能很好地去除基质干扰。从 6 种选定的印度产品中的 30 个样品和 NIST SRM 1849a 中每个样品的测定值的相对标准偏差均小于 20%。加标回收样品的平均回收率在 91.75%至 101.14%之间。
方法 2011.10 满足了 AOAC SPIFAN 规定的标准方法性能要求。因此，我们建议采用方法 2011.10 作为 BIS 分析“印度”婴儿配方和儿科配方及成人营养品中维生素 B12 的官方方法。
这是 AOAC 印度分部承担的第一个 SLV 项目，旨在通过验证其在“印度”婴儿配方和儿科配方及成人营养品中的适用性，扩展 AOAC 方法 2011.10 测定维生素 B12 的范围。
