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目的:探讨天麻熄风胶囊的DNA条形码鉴定方法与价值。方法:2015年1月到11月,选择3批天麻熄风胶囊(批号20150918、200150919、20151019)进行显微鉴定与DNA条形码鉴定,选择的基因序列为天麻素线粒体A基因,进行基因测序与遗传分析。结果:显微鉴定显示天麻细胞无色,呈现长卵形、长椭圆形或类圆形,遇碘液显棕色;蔓荆子非腺毛2-3个细胞,顶端细胞基部稍粗;甘草纤维成束、壁厚,周围薄壁细胞可形成晶纤维;半夏椭圆形细胞存在于粘液细胞中。3批天麻熄风胶囊的天麻素线粒体A基因特定序列中,A含量为27.9%、C为24.9%、G为18.3%、T为28.9%,G-T含量明显高于G/C含量(43.2%)(P<0.05)。遗传分析显示20150918批次的种间最小K-2P距离为0.006,种间最大K-2P距离为0.241;20150919批次种间最小K-2P距离为0.008,最大K-2P距离为0.074;20151019批次的种间最小K-2P距离为0.008,种间最大K-2P距离为0.133。结论:天麻熄风胶囊成分复杂,显微鉴定技术存在一定的缺陷,DNA条形码鉴定能有效反映胶囊成分,且能很好判定药物批次的种内与种间变异,具有很好的鉴别价值。
Objective: To explore the method and value of DNA barcode identification of Tianma Xifeng Capsule. METHODS: From January to November 2015, three batches of Tianma Xifeng Capsules (batch numbers 20150918, 200150919, 20151019) were selected for microscopic identification and DNA barcode identification. The selected gene sequence was Gastrodin A mitochondrial gene, and gene sequencing and genetic analysis analysis. Results: The microscopic identification showed that the cells of Gastrodia elata were colorless, oval, oval or round in shape, brown in case of iodine solution; 2-3 cells of non-glandular hairy, , Wall thickness, the surrounding parenchyma cells can form crystal fibers; Pinellia oval cells exist in mucus cells. The content of A was 27.9%, C was 24.9%, G was 18.3%, T was 28.9%, GT content was significantly higher than that of G / C (43.2%) in the gastrodin A gene specific sequence of Tianma Xifeng capsule. (P <0.05). Genetic analysis showed that the minimum K-2P distance between species in batch 20150918 was 0.006, the maximum K-2P distance between species was 0.241; the minimum K-2P distance between batch 20150919 was 0.008 and the maximum K-2P distance was 0.074; The smallest inter-species K-2P distance was 0.008, and the largest K-2P distance was 0.133. Conclusion: The composition of Tianma Xifeng Capsule is complex, and the microscopic identification technique has some defects. DNA barcode identification can effectively reflect the composition of the capsule, and can well determine the intraspecific and interspecific variation of drug batches, which has a good identification value.