基于黄芩苷单克隆抗体的ELISA快速检测方法的建立

目的: 建立黄芩苷快速灵敏的的免疫分析检测方法。 方法: 以制备出的黄芩苷特异性单克隆抗体为基础,选择单抗最佳工作浓度,建立黄芩苷间接竞争酶联免疫分析方法,并应用此方法检测精制清开灵注射液中的黄芩苷含量。 结果: 该方法线性范围为2.67~1029.00 ng·mL^-1,组内和组间差异均不超过3.31%,平均回收率为100.9%,检测工作时间可控制在4 h内。采用该方法检测精制清开灵注射液中黄芩苷的含量,所得结果与HPLC一致。 结论: 建立了黄芩苷的ELISA方法,可为含黄芩苷的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 the determination of baicalin(BA). Methods: Indirect competitive ELISA was developed by using anti-BA monoclonal antibody(anti-BA MAb),and then it was applied to BA measurement in the traditional Chinese medicine injection Jingzhi Qingkailing. Results: The standard curve of established ELISA was linear between 2.67 and 1029.00 ng·mL^-1.The average recovery was 100.9%,and the relative standard deviation(RSD) of measurements was <3% in the intra-assay,and <4% in the inter-assay.The test of samples could be finished in 4hs by using this ELISA method.The analysis result of ELISA method was consistent with that of HPLC test in BA determination of Jingzhi Qingkailing injection. Conclusion: Quick ELISA method for the BA determination is well established and can be applied to the quality control of traditional Chinese herbal medicine/compound preparation which contains baicalin.