ELISA法测定半夏及其炮制品中半夏凝集素含量  

Determination of Pinellia ternata lectin in Pinelliae Rhizoma and its processed products by ELISA

作　　者：郁红礼[1,2,3] 谢雨薇 吴皓 王迅[4] 刘冰冰 崔小兵 张萍 许艳青[1] YU Hongli;XIE Yuwei;WU Hao;WANG Xun;LIU Bingbing;CUI Xiaobing;ZHANG Ping;XU Yanqing(Pharmacology College of Nangjing University of Chinese Medicine,Nangjing 210023,China;Jiangsu Key Laboratory of Chinese Medicine Processing,Nangjing 210023,China;Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing,Nangjing 210023,China;Jiangsu Province Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nangjing 210029,China;National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]南京中医药大学药学院,南京210023 [2]江苏省中药炮制重点实验室,南京210023 [3]中华人民共和国教育部中药炮制规范化及标准化工程研究中心,南京210023 [4]南京中医药大学附属医院江苏省中医院,南京210029 [5]中国食品药品检定研究院,北京100050

出　　处：《中国药品标准》2023年第3期268-274,共7页Drug Standards of China

基　　金：国家重点研发计划项目(2018YF1707000);国家自然科学基金项目(81173549)。

摘　　要：目的:研究双抗夹心酶联免疫ELISA法作为半夏及其炮制品中毒蛋白半夏凝集素蛋白(Pinellia ternata lectin,PTL)检查方法的可行性。方法:采用双抗夹心ELISA法,测定半夏不同炮制品中PTL含量。结果:方法学考察结果显示,非线性四参数曲线拟合相关系数r^(2)=0.9999,表明当PTL浓度在0.075~4.80 ng•mL^(-1)浓度范围内,以非线性四参数拟合曲线计算PTL含量的准确度高,且方法精密度、稳定性、重复性较好。26批半夏生品及75批半夏炮制品(清半夏、法半夏、姜半夏各25批)中半夏中PTL含量均值为30.9 mg•g^(-1),其炮制品清半夏、法半夏、姜半夏中PTL含量均值分别为0.92 mg•g^(-1),48.80μg•g^(-1),130.32 ng•g^(-1),表明炮制后PTL含量显著下降,姜半夏中PTL含量最低,其次为法半夏,清半夏含量最高。同时建立半夏不同饮片中内源性毒性蛋白凝集素蛋白限度检查方法,建议清半夏、法半夏、姜半夏中PTL含量分别不超过2.70 mg•g^(-1)、160.0μg•g^(-1)、510.0 ng•g^(-1)。结论:ELISA法测定半夏凝集素蛋白准确可靠,且成本低,可快速批量测定,可以作为半夏及其炮制品中PTL检查方法。Objective:To study the feasibility of using double antibody sandwich enzyme linked immunosorbent assay(ELISA)as a method for the detection of Pinellia ternata lectin(PTL)in Pinelliae Rhizoma(PR)and its processed products.Methods:Double antibody sandwich ELISA was adopted for determinationt of PTL in processed PR products.Results:The results of methodological investigation showed that the correlation coefficient r^(2) of non-linear four parameter curve fittiwas 0.9999.This result indicated that when the concentration of PTL was within the range of 0.075-4.80 ng•mL^(-1),the accuracy of calculating PTL content using non-linear four parameter curve fitting was high.The method had good precision,stability,and repeatability.6 batches of raw PR,25 batches of Pinelliae Rhizoma Praeparatum Cum Alumine(PRPCA),25 batches of Pinelliae Rhizoma Praeparatum(PRP)and 25 batches of Pinelliae Rhizoma Praeparatum Cum Zingibere Et Alumine(PRPCAEA)were collected,and the content determination results showed that the average PTL content in raw PR was 30.9 mg•g^(-1),and the average PTL content in processed products of PRPCA,PRP and PRPCAEA were 0.92 mg•g^(-1),48.80μg•g^(-1),130.32 ng•g^(-1),respectively,indicating a significant decrease in PTL content after processing.The content of PTL in PRPCAEA was the lowest,followed by PRP,and PRPCA was the highest.At the same time,this paper established a limit test method for endogenous toxic protein PTL in prepared slices of PR.It was recommended that the content of PTL in PRPCA,PRP and PRPCAEA should not exceed 2.70 mg•g^(-1),160.0μg•g^(-1),510.0 ng•g^(-1),respectively.Conclusion:The ELISA method for the PTL determination is accurate and reliable.It has low cost and can be quickly measured in batchescan.ELISA can be used as a method for the detection of toxic protein PTL in PR and its processed products.

关 键 词：半夏 半夏凝集素 有毒物质控制 含量测定 ELISA 炮制 

分 类 号：R921.2[医药卫生—药学]