基于DNA条形码及HPLC对市售刺五加的种质资源鉴定和质量评价  

Identification and quality evaluation of germplasm resources of commercial Acanthopanax senticosus based on DNA barcodes and HPLC

作　　者：刘珊瑚 张志飞 黄钰莹 刘子齐 陈文沁 阿木拉哈 王馨 石玥 张晓芹[1] 何高洁 安克露 王晓晖 魏胜利[1,3] LIU Shan-hu;ZHANG Zhi-fei;HUANG Yu-ying;LIU Zi-qi;CHEN Wen-qin;AMU La-ha;WANG Xin;SHI Yue;ZHANG Xiao-qin;HE Gao-jie;AN Ke-lu;WANG Xiao-hui;WEI Sheng-li(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Modern Research Center for Traditional Chinese Medicine,Beijing Institute of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 102488,China;Engineering Research Center of Good Agricultural Practice for Chinese Crude Drugs of Ministry of Education,Beijing 100102,China;Heilongjiang Beicaotang Traditional Chinese Medicine Co.,Ltd.,Harbin 150900,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]北京中医药大学北京中医药研究院中药现代研究中心,北京102488 [3]中药材规范化生产教育部工程研究中心,北京100102 [4]黑龙江北草堂中药材有限责任公司,黑龙江哈尔滨150900

出　　处：《药学学报》2024年第7期2171-2178,共8页Acta Pharmaceutica Sinica

基　　金：兴安盟科技计划项目(基于分子防伪技术的精准药材北柴胡选育及栽培技术研究,20221156);企业横向课题(桔梗等4种精准药材批次分子防伪技术研究项目,2020110031009385);国家重点研发计划项目(中医药现代化专项,2022YFC3501505)。

摘　　要：刺五加Acanthopanax senticosus(Rupr.et Maxim.)Harms是我国东北道地药材之一,本研究根据之前的叶绿体基因组测序结果,选取atpI和atpB_rbcL基因片段对市售刺五加样品进行种质资源鉴定,并利用HPLC对其紫丁香苷含量进行测定,以期确定市售刺五加的主流种质和质量。本研究从24省47市收集80份刺五加样品并提取DNA,通过PCR扩增atpI和atpB_rbcL基因片段,分析结果表明,2个基因片段联合分析形成7个单倍型(H2、H5、H8、H10、H12、H14、H23),其中来自黑龙江伊春、哈尔滨尚志、黑龙江绥化、辽宁抚顺、辽宁本溪、吉林长白和吉林靖宇的单倍型H2是主流单倍型,占总样本量的58.75%。H14和H23为产地独有单倍型,分别来自黑龙江省双鸭山市饶河县和黑龙江省绥化市明水县。通过HPLC检测市售刺五加样品中的紫丁香苷含量,结果表明73.96%样品紫丁香苷含量符合药典标准。样品间的紫丁香苷含量差异显著,在0.0037%~0.5245%之间,相差0.5208%,说明刺五加市场样品质量参差不齐,但是不同单倍型之间市售刺五加紫丁香苷含量没有显著性差异。市场样品中单倍型H23中的紫丁香苷含量比较高,因此可能是质量比较好的种质。本文对刺五加市场样品的种质资源和药材质量进行研究,有利于指导刺五加市场样品流通、临床合理用药和后续优良品种筛选。Acanthopanax senticosus is one of the genuine regional herb in Northeast China.In this study,we identified the germplasm resources of commercial A.senticosus samples based on atpI and atpB_rbcL according to the previous chloroplast genome sequencing results,and determinated the content of syringin by HPLC to evaluated the quality of commercial samples.A total of 80 A.senticosus samples were collected from 47 cities in 24 provinces.DNA was extracted to amplify the products of atpI and atpB_rbcL by PCR.The results showed that 7 haplotypes(H2,H5,H8,H10,H12,H14,H23)were formed by the combined analysis of the two gene fragments.H2 from Yichun in Heilongjiang,Shangzhi in Harbin,Suihua in Heilongjiang,Fushun in Liaoning,Benxi in Liaoning,Changbai in Jilin and Jingyu in Jilin were the dominant genotype,representing 58.75%of the total samples.H14 and H23 were the unique haplotypes of the producing area.It is speculated that the commercial A.senticosus samples with haplotypes of H14 and H23 are from Raohe,Shuangyashan,Heilongjiang and Mingshui,Suihua,Heilongjiang,respectively.HPLC analysis indicated that the content of syringin in 73.96%of the samples met the Pharmacopoeia standards.There was a significant difference in the content of syringin among the samples,ranging from 0.0037%to 0.5245%,with a difference of 0.5208%,indicating that the quality of the samples in the market of A.senticosus was uneven.However,there were no significant differences in the contents of syringin in the commercial A.senticosus among different haplotypes.The content of syringin in the haplotype H23 in the market sample is relatively high,so it may be a germplasm with better quality.This study researched the germplasm resources and medicinal materials quality of commercial A.senticosus samples and will help guide the commercial circulation,reasonable medication of A.senticosus,and the screening of excellent germplasm.

关 键 词：刺五加 DNA条形码 分子鉴定 紫丁香苷 质量评价 

分 类 号：R931[医药卫生—生药学]