College of Pharmacy, Research Center for Traditional Chinese Medicine of Lingnan (Southern China), National Engineering Research Center for Modernization of Traditional Chinese Medicine Lingnan Resources Branch, Jinan University, Guangzhou, Guangdong, China.
Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization (2021B1212040007), Guangzhou, Guangdong, China.
PLoS One. 2022 Aug 12;17(8):e0273021. doi: 10.1371/journal.pone.0273021. eCollection 2022.
Donkey-hide gelatin is an important traditional Chinese medicine made from donkey skin. Despite decades of effort, identifying the animal materials (donkeys, horses, cattle and pigs) in donkey-hide gelatin remains challenging. In our study, we aimed to identify marker peptides of donkey-hide gelatin and its adulterants and develop a liquid chromatography-tandem mass spectrometry method to identify them. Theoretical marker peptides of four animals (donkeys, horses, cattle and pigs) were predicted and verified by proteomic experiments, and 12 species-specific marker peptides from donkey-hide gelatin and its adulterants were identified. One marker peptide for each gelatin was selected to develop the liquid chromatography-tandem mass spectrometry method. The applicability of the method was evaluated by investigating homemade mixed gelatin samples and commercial donkey-hide gelatin products. Using the liquid chromatography-tandem mass spectrometry method, the addition of cattle-hide gelatin and pig-hide gelatin to donkey-hide gelatin could be detected at a level of 0.1%. Horse-hide gelatin was detected when added at a level of 0.5%. Among 18 batches of donkey-hide gelatin products, nine were identified as authentic, and eight of the remaining samples were suspected to be adulterated with horse materials. These results provide both a practical method to control the quality of donkey-hide gelatin and a good reference for quality evaluations of other medicinal materials and foods containing protein components.
阿胶是一种由驴皮制成的重要中药。尽管经过几十年的努力，鉴定阿胶中的动物材料（驴、马、牛和猪）仍然具有挑战性。在我们的研究中，我们旨在鉴定阿胶及其掺杂物的标记肽，并开发一种液相色谱-串联质谱法来识别它们。通过蛋白质组学实验预测和验证了四种动物（驴、马、牛和猪）的理论标记肽，并鉴定了 12 种来自阿胶及其掺杂物的种特异性标记肽。为每种明胶选择一个标记肽来开发液相色谱-串联质谱法。通过研究自制混合明胶样品和商业阿胶产品评估了该方法的适用性。使用液相色谱-串联质谱法，可以在 0.1%的水平检测到牛皮胶和猪皮胶添加到阿胶中的情况。当添加马皮胶时，可以检测到 0.5%的水平。在 18 批阿胶产品中，有 9 批被鉴定为正品，其余 8 批样品疑似掺有马材料。这些结果不仅为控制阿胶质量提供了一种实用方法，而且为其他含有蛋白质成分的药材和食品的质量评价提供了很好的参考。
