Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing, 210023, PR China; School of Pharmacy and Department of Chemistry, University of Wisconsin-Madison, Madison, WI, 53705, USA; Jiangsu Key Laboratory of Research and Development in Marine Bio-resource Pharmaceutics, Nanjing University of Chinese Medicine, Nanjing, 210023, PR China; School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, PR China.
Guizhou Guangjitang Pharmaceutical Co., Ltd., Guiyang, 550014, PR China.
Anal Chim Acta. 2019 Dec 27;1092:32-41. doi: 10.1016/j.aca.2019.09.064. Epub 2019 Sep 26.
Deer-hide gelatin (DHG) is an important animal-derived traditional Chinese medicine (TCM), which has been applied in TCM for over 400 years. However, it is extremely difficult to distinguish DHG with adulteration or made with other animal skins due to the highly processing procedure. Therefore, a simple strategy for identifying species-specific peptide biomarkers in deer-hide gelatin (DHG) is needed. In the present study, untargeted and targeted mass spectrometry approaches were implemented to analyze comprehensive peptidomic profiles of trypsin-digested animal gelatins. Mathematics set theory was then used to interrogate the relationship between different samples and peptides in the target species set, while the peptides were not considered as species-specific biomarkers in other sets. Two peptides were identified as DHG-specific peptides. Targeted mass spectrometry approach was then used to verify these two peptides. It showed that these two peptides could be used for distinguishing DHG from other animal hide gelatins. The present strategy provides a simple method for peptide biomarker discovery, which can be applied in the identification of specific peptides in some highly processed animal derived traditional Chinese medicines (TCMs). Thus, the present work provides an effective strategy for rapid, simple discovery and application of species-specific peptide biomarkers to ensure animal derived TCMs quality and make them authenticable and traceable.
鹿皮明胶（DHG）是一种重要的动物源性传统中药（TCM），已经在中医药中应用了 400 多年。然而，由于其高度加工的过程，极难区分掺假或用其他动物皮制成的 DHG。因此，需要一种简单的策略来识别鹿皮明胶（DHG）中的物种特异性肽生物标志物。在本研究中，采用非靶向和靶向质谱方法分析了胰蛋白酶消化的动物明胶的综合肽组学图谱。然后使用数学集合论来询问目标物种集中不同样本和肽之间的关系，而这些肽在其他集中不被视为物种特异性生物标志物。鉴定出两个肽作为 DHG 特异性肽。然后使用靶向质谱方法验证了这两个肽。结果表明，这两个肽可用于区分 DHG 与其他动物皮明胶。本策略提供了一种简单的肽生物标志物发现方法，可应用于某些高度加工的动物源性传统中药（TCM）中特定肽的鉴定。因此，本工作为快速、简单地发现和应用物种特异性肽生物标志物提供了一种有效的策略，以确保动物源性 TCM 的质量，并使其可验证和可追溯。
