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目的建立并完善从中成药及保健品中提取DNA的有效方法,并在人参类制剂中进行验证。方法使用中成药和保健品中常用的辅料(如淀粉、蔗糖、葡萄糖等)模拟CTAB法提取DNA过程,用以筛选影响提取DNA效率的因素;使用甲醇作为沉淀试剂优化传统的CTAB法,并使用MAS-PCR方法对市场上人参类中成药及保健品进行分子鉴定,以验证优化的技术。结果中成药及保健品中的淀粉(包括药源性淀粉和辅料添加的淀粉)通过CTAB反应后生成极性较大的复合物,其与70%乙醇不能互溶,而且在碱性条件下对DNA有强烈的溶解作用,从而影响了CTAB法提取DNA的提取效率;利用甲醇作为沉淀试剂能够有效去除CTAB裂解过程中产生的高极性复合物的影响,70%甲醇-醋酸钠溶液能够有效提取中成药及保健品的DNA;利用优化的DNA提取方法和MAS-PCR法实现了11种市售人参类制剂的快速分子鉴定,结果显示8种与商品描述一致、1种有掺伪情况、2种为伪品。结论使用甲醇作为沉淀试剂优化传统的CTAB法,可以较好地克服中药制剂中淀粉对DNA提取的不利影响,可以有效地实现基于DNA的中药制剂分子鉴定。
Objective To establish and improve an effective method for extracting DNA from Chinese patent medicines and health products and to verify it in ginseng preparations. Methods CTAB method was used to extract the DNA from traditional Chinese medicine and health care products, such as starch, sucrose and glucose, to screen the factors affecting DNA extraction efficiency. Methanol was used as a precipitation reagent to optimize the traditional CTAB method. MAS-PCR method on the market of Chinese medicines ginseng and health products molecular identification to verify the optimization of technology. Results The starch in traditional Chinese medicine and health products (including starches added with drug-derived starches and excipients) produced a more polar complex through CTAB reaction, which could not be miscible with 70% ethanol, and under alkaline conditions, DNA Which can affect the extraction efficiency of DNA extracted by CTAB method. The use of methanol as a precipitating reagent can effectively remove the influence of high polarity compounds produced during CTAB lysis, and the 70% methanol-sodium acetate solution can be effectively extracted Medicine and health care products DNA; Rapid molecular identification of 11 kinds of commercially available ginseng preparations by using optimized DNA extraction method and MAS-PCR method, the results showed that 8 kinds of goods with the same description, a kind of adulteration, two To counterfeit goods. Conclusion Methanol as a precipitation reagent to optimize the traditional CTAB method can better overcome the unfavorable effects of starch on DNA extraction in traditional Chinese medicine preparations and can effectively realize the molecular identification of traditional Chinese medicine preparations based on DNA.