Biofarma Group Srl, Via Castelliere 2, Mereto di Tomba, 33036 Udine, Italy.
Institute of Pathology Department of Medicine, University of Udine, 33100 Udine, Italy.
Molecules. 2021 Mar 12;26(6):1566. doi: 10.3390/molecules26061566.
Alpha-galactosidase (α-Gal) is an enzyme responsible for the hydrolyzation of glycolipids and glycoprotein commonly found in dietary sources. More than 20% of the general population suffers from abdominal pain or discomfort caused by intestinal gas and by indigested or partially digested food residuals. Therefore, α-Gal is used in dietary supplements to reduce intestinal gases and help complex food digestion. Marketed enzyme-containing dietary supplements must be produced in accordance with the Food and Drug Administration (FDA) regulations for Current Good Manufacturing Practice (cGMPs).
in this work we illustrated the process used to develop and validate a spectrophotometric enzymatic assay for α-Gal activity quantification in dietary supplements.
The validation workflow included an initial statistical-phase optimization of materials, reagents, and conditions, and subsequently a comparative study with another fluorimetric assay. A final validation of method performance in terms of specificity, linearity, accuracy, intermediate-precision repeatability, and system precision was then executed.
The proven method achieved good performance in the quantitative determination of α-Gal activity in commercial food supplements in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals (ICH) guidelines and is suitable as a rapid in-house quality control test.
α-半乳糖苷酶（α-Gal）是一种负责水解糖脂和糖蛋白的酶，这些物质通常存在于饮食来源中。超过 20%的普通人群会因肠道气体和未消化或部分消化的食物残渣引起腹痛或不适。因此，α-Gal 被用于膳食补充剂中，以减少肠道气体并帮助复杂食物消化。市售的含酶膳食补充剂必须按照食品和药物管理局（FDA）的现行良好生产规范（cGMP）进行生产。
在这项工作中，我们展示了用于开发和验证用于定量测定膳食补充剂中α-Gal 活性的分光光度酶法的过程。
验证工作流程包括对材料、试剂和条件进行初始统计阶段优化，然后与另一种荧光法进行比较研究。最后，根据国际人用药品注册技术协调会（ICH）指南对方法性能进行特异性、线性、准确性、中间精密度重复性和系统精密度的最终验证。
该方法在定量测定商业食品补充剂中的α-Gal 活性方面表现良好，符合 ICH 指南，适合作为快速内部质量控制测试。
