A novel strategy for the discrimination of gelatinous Chinese medicines based on enzymatic digestion followed by nano-flow liquid chromatography in tandem with orbitrap mass spectrum detection.
A novel strategy for the discrimination of gelatinous Chinese medicines based on enzymatic digestion followed by nano-flow liquid chromatography in tandem with orbitrap mass spectrum detection.
复制标题
基于酶消化、纳流液相色谱和轨道阱质谱检测的凝胶状中药鉴别新策略
DOI：
10.2147/ijn.s82291
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发表时间：
2015
影响因子：
8
通讯作者：
Tam JP
中科院分区：
文献类型：
--
作者：
Yang H;Shen Y;Xu Y;Maqueda AS;Zheng J;Wu Q;Tam JP
由哺乳动物的皮肤、角或爬行动物壳制成的胶状中药是一种非常重要的动物源性中药。它们被广泛用作造血和止血药物，用于中医诊所治疗眩晕、心悸、血尿和失眠；作为老年人或妇女分娩后等体弱者的流行补品；或进一步加工成某些人群的保健品。然而，仅用《中国药典》中的常规方法不能区分它们，因为缺乏足够的专属性，因此，即使添加被测成分也能满足要求。本研究以细辛、鹿茸、龟甲三种凝胶型中药材为研究对象，提出了一种基于酶消化-纳米流动相色谱-Orbitrap-MS联用的新方法。从这些药物的消化中确定的14个诊断片段被专门选择用于区分它们。利用这一策略的优势，可以方便、可行地鉴定每种药材中的标志性蛋白来源的酶消化肽，从而可以作为其原料药材形式的潜在生物标志物，特别是粉末状和复合体，这是鉴定目的的直接依据。
Gelatinous Chinese medicines made from mammalian skin or horn or reptile shell are a very important type of animal-derived Chinese medicine. They have been extensively used either as both hemopoietic and hemostatic agents to treat vertigo, palpitation, hematuria, and insomnia in traditional Chinese medicine clinics; consumed as a popular tonic for weaker persons such as the elderly or women after giving birth; or further manufactured to health supplements for certain populations. However, they cannot be discriminated from each other by only using the routine approach in the Chinese Pharmacopoeia, as it lacks enough specificity and, consequently, and the requirements can be met even by adding assayed ingredients. In this study, our efforts to differentiate three gelatinous Chinese medicines, Asini Corii Colla, Cervi Cornus Colla, and Testudinis Carapacis ET Plastri Colla, are presented, and a novel strategy based on enzymatic digestion followed by nano-flow liquid chromatography in tandem with orbitrap mass spectrum detector analysis is proposed herein. Fourteen diagnostic fragments identified from the digests of these medicines were exclusively selected for their discrimination. By taking advantage of the favorable features of this strategy, it is feasible and convenient to identify enzymatic-digested peptides originated from signature proteins in each medicine, which thus could be employed as potential biomarkers for their form of raw medicinal material, and the pulverized and the complex especially, that being the direct basis for authentication purpose.