免疫亲和法敲除三七总皂苷中三七皂苷R1的实验研究  

Experimental Study on Knock-Out of Notoginsenoside R1 from Notoginseng Total Saponins by Immunoaffinity Chromatography

作　　者：邵晓[1,2] 胡超 林子璇 邵建国[5] 赵玉男[2] SHAO Xiao;HU Chao;LIN Zixuan;SHAO Jianguo;ZHAO Yunan(Nantong Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Nantong 226001,Jiangsu,China;School of Medicine&Holistic Integrative Medicine of Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China;Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,Henan,China;The Third Nantong Hospital Affiliated to Nantong University,Nantong 226001,Jiangsu,China)

机构地区：[1]南京中医药大学南通附属医院,江苏南通226001 [2]南京中医药大学医学院·整合医学学院,江苏南京210023 [3]南京中医药大学,江苏南京210023 [4]河南中医药大学第一附属医学院,河南郑州450000 [5]南通大学附属南通第三医院,江苏南通226001

出　　处：《中华中医药学刊》2022年第6期70-73,I0029-I0030,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81873025);江苏省卫生厅基金(H2017057);江苏省研究生科研与实践创新计划(KYCX18-1567)。

摘　　要：目的基于目标成分敲除技术,尝试免疫亲和法敲除三七总皂苷中的三七皂苷R1(notoginsenoside R1),并与制备液相法进行比较。方法首先,运用制备液相色谱系统,基于目标成分三七皂苷R1制定洗脱程序,回收敲除三七皂苷R1后的三七皂苷样品;其次,制备三七皂苷R1-白蛋白半抗原抗体,并采用该抗体制备免疫亲和柱,利用目标成分特异亲和吸附原理敲除三七总皂苷中的三七皂苷R1,并回收敲除掉目标成分后的样品;最后,采用超高效液相色谱-电喷雾检测器(UPLC-CAD)柱后补偿结合内参法,对敲除目标成分后的三七皂苷样品进行成分分析。结果UPLC-CAD成分分析显示,三七总皂苷中原人参二醇型皂苷(PPDs)和原人参三醇型皂苷(PPTs)含量的比值为0.67,Rb1/Rg1的值为0.55。制备液相法能够从三七总皂苷中彻底敲除三七皂苷R1,敲除后PPDs和PPTs含量的比值为0.70,Rb1/Rg1的值为1.82。免疫亲和法可以敲除大部分三七皂苷R1,敲除后PPDs和PPTs含量的比值为0.53,Rb1/Rg1的值为0.33。结论两种敲除方法均可以将三七皂苷R1从三七总皂苷内敲除,但同时也会改变总皂苷中各单体皂苷成分含量的比值。对于总皂苷中Rb1/Rg1的值,免疫亲和法影响小于制备液相法。Objective Based on the target component knock-out technology,this research group tried to knock out notoginsen-oside R1 by immunoaffinity chromatography and compared it with liquid polymerization.Methods Firstly,the preparation liquid chromatography system was used to formulate the elution procedure based on the target component Panax notoginseng saponin R1,and recover the Panax notoginseng saponin samples after knocking out Panax notoginseng saponin R1.Secondly,Panax notogin-seng saponin R1 albumin hapten antibody was prepared,and the antibody was used to prepare the immune affinity column.The Panax notoginseng saponin R1 in Panax notoginseng total saponins was knocked out by using the principle of specific affinity ad-sorption of target components,and the samples after knocking out target components were recovered.Finally,the content of 37 saponin samples after knocking out target components was analyzed by ultra performance liquid chromatography electrospray de-tector(UPLC-CAD)post column compensation and internal reference method.Results UPLC-CAD component analysis showed that the ratio of protopanaxadiol type saponins(PPDs)and protopanaxatriol type saponins(PPTs)in the total saponins of notoginseng was 0.67,and the value of Rb1/Rg1 was 0.55.The preparative liquid method can completely knock out the notogin-senoside R1 from the total saponins of notoginseng.The ratio of PPDs to PPTs content after knock-out was 0.70,and the value of Rb1/Rg1 was 1.82.The immunoaffinity chromatography can knock out most of notoginsenoside R1,the ratio of PPDs and PPTs content after knoc-kout was 0.53,and the value of Rb1/Rg1 was 0.33.Conclusion Both knock-out methods can not on-ly knock out the notoginsenoside R1 from the total saponins of notoginseng,but also change the ratio of the content of the monomer saponins in the total saponins.For the value of Rb1/Rg1 in total saponins,the immunoaffinity column chromatography has less influence than the preparative liquid method.

关 键 词：三七 三七皂苷R1 免疫亲和法 制备液相 UPLC-CAD 

分 类 号：R282.71[医药卫生—中药学]