胆粉(汁)类药材的DNA指纹图谱鉴别方法研究

《中国药典》收载了猪胆粉、牛胆粉、蛇胆汁、羊胆、鹅胆粉、熊胆粉等药材。胆粉(汁)类药材在我国药用历史悠久,疗效明确。但其形态相似、多种药材缺乏专属性显微鉴别特征和化学特征,存在混伪风险,尤其是对正品中掺杂伪品的情况检出困难。控制胆粉类中药质量,保证临床疗效,解决胆粉类药材的真伪优劣鉴别问题是关键。STR分型技术是通过PCR扩增长度差异分型,比较鉴定生物个体的一种方法。该研究基于STR分型原理建立了一种简单快捷,可用于胆粉类药材及其掺杂样品的通用DNA指纹图谱鉴别方法,并收集猪、牛、羊、鸡、鸭、鹅、蛇、熊、鱼等原动物或胆粉样本,使用筛选获得的鉴别引物12S-L1091/12S-H1478和16S-L3428/16S-H3667扩增并进行STR分型获得DNA指纹图谱。结果各物种的STR指纹图谱不同,可进行物种鉴别,且DNA指纹图谱上能同时反映正品和伪品的信息,可鉴别正伪掺杂品。因此,通过2个引物的结合,完成了胆粉类药材及其正伪掺杂品的鉴别。此外,该研究建立的DNA指纹图谱鉴别方法也可用于其他动物药材的真伪鉴别和掺杂品检出。

DNA fingerprinting identification of bile power(bile)medicines

The pig bile powder,bovine bile powder,snake bile,sheep bile,goose bile powder,and bear bile powder were contained by the Chinese Pharmacopoeia.The bile power medicine has a long history in traditional Chinese medicine and definite effect.However,the medicine of bile powder(bile)are similar in morphology.Besides,many medicine lack specific microscopic identification characteristics and chemical characteristics.There is a risk of adulteration,especially when the fake medicine were mixed in authentic medicine,it is difficult to detection.The key to control the quality and ensures the clinical efficacy is the good or bad,true or false of the bile power medicine.The STR typing technology is a method that according to differential typing of PCR amplified lengths to compare and identify individual organisms.Based on the principle of STR typing,the easily,rapid DNA fingerprinting method to identify the bile power and adulteration was established.The original animal or bile powder of pigs,cattle,sheep,chickens,ducks,geese,snakes,bears,fish were collected,the 12 S-L1091/12 S-H1478 and 16 S-L3428/16 S-H3667 was obtained by sifted,the DNA fingerprinting of the bile power and adulteration was obtained by STR typing.Every species has different STR fingerprints,so different species can be identified.Besides,the fingerprints have both the authentic and fake′s information,the adulteration of authentic and fake can be identified.Therefore,the method to identify the bile power and adulteration was achieved through the combination of two primers.The DNA fingerprinting method established in this study can also be used for other animal medicine.