基于芍药苷单克隆抗体的ELISA快速检测方法建立  

Establishment of quick enzyme-linked immunosorbent assay with monoclonal antibody against paeoniflorin

作　　者：屈保平[1] 冯红 续洁琨[1] 屈会化[3] 孔慧[1] 王雪茜[1] 王庆国[1] 赵琰[1] 

机构地区：[1]北京中医药大学基础医学院,北京100029 [2]天士力中药资源研究中心,天津300410 [3]北京中医药大学北京中医药研究院,北京100029

出　　处：《药物分析杂志》2016年第11期1931-1935,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：国家自然科学基金(81573573)

摘　　要：目的:建立芍药苷快速、灵敏的酶联免疫分析(ELISA)检测方法,并使用新方法检测中药材白芍中芍药苷的含量。方法:以制备出的芍药苷特异性单克隆抗体为基础,通过考察线性关系、精密度、回收率等方法学指标,建立芍药苷间接竞争酶联免疫分析(icELISA)方法,并应用此方法检测中药材白芍中的芍药苷含量。结果:该方法线性范围为3.9~250 ng·mL^(-1),板内和板间差异均不超过9.8%,平均回收率<110%,检测工作时间可控制在2 h内。采用该方法检测中药材白芍中芍药苷的含量,所得结果与HPLC一致。结论:建立的芍药苷icELISA方法,可为含芍药苷的微量生物样品检测、中药材及复方的质量控制分析提供更加快速灵敏的检测方法。Objective. To establish a quick enzyme-linked immunosorbent assay （ ELISA ） method for thedetermination of paeoniflorin （ PF ）. Methods： Indirect competitive ELISA （ icELISA ） was developed by using anti,PF monoclonal antibody （ anti-PF MAb ）. The linearity, accuracy, the average recovery and the likewere detected. Then the established method was applied to PF measurement in the traditional Chinese medicine Paeoniae Radix Alba. Results： The standard curve of established icELISA was linear between 3.9-250 ng. mL-1, the average recovery was 〈110%, and the relative standard deviation （ RSD ） of measurements was 〈 9.8% intraassay and inter-assay. The test of samples could be finished in 2 h by using this icELISA method. The analysis result of ELISA method was consistent with HPLC test in PF determination of Chinese medicine Paeoniae Radix Alba. Conclusion： A quick icELISA method for the PF determination is well established, which can be used for the trace biological sample detection, as well as the quality control of traditional Chinese herbal medicine/ compound preparation concerning paeoniflorin.

关 键 词：芍药苷 单萜类糖苷化合物 白芍 单克隆抗体 快速含量测定 微量生物样品检测 酶联免疫分析 

分 类 号：R917[医药卫生—药物分析学]