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目的:建立免疫分析技术对麻杏石甘汤中甘草酸(GA)的含量分析方法。方法:采用杂交瘤技术制备了甘草酸的单克隆抗体,采用竞争性酶联免疫吸附测定(ELISA)技术,同时结合HPLC色谱分析方法(采用Phenomsil C18(4.6mm×250 mm,5μm),流动相为甲醇-0.2 mol/L醋酸胺溶液-冰醋酸(67∶33∶1),检测波长为250 nm,对复方中药“麻杏石甘汤”中的甘草酸含量进行测定。结果:麻杏石甘汤中成分GA分别在0.3125~10μg(r=0.9998)范围内线性关系良好,平均回收率分别为109.0%、92.5%和103.9%,3批样品中GA含量均值为9.3003 mg·g-1;与HPLC测定结果相比较结果相近,大多<±5%范围。结论:该方法准确可靠,重复性好,对于HPLC测定提供了有益的技术补充。
Objective: To establish an immunoassay technique for the determination of glycyrrhizic acid (GA) in Maxing Shigan Decoction. METHODS: Monoclonal antibodies against glycyrrhizin were prepared by hybridoma technique. The products were characterized by competitive enzyme-linked immunosorbent assay (ELISA) and HPLC (Phenomicil C18 (4.6 mm × 250 mm, 5 μm) The content of glycyrrhizic acid in compound Chinese medicine “Maxing Shigan Decoction” was determined by the method of methanol-0.2 mol / L ammonium acetate solution-acetic acid (67:33:1) and the detection wavelength was 250 nm.Results: The GA of Xingshiganan decoction has a good linear relationship in the range of 0.3125 ~ 10μg (r = 0.9998), with the average recoveries of 109.0%, 92.5% and 103.9%, respectively. The average GA content of three batches of samples was 9.3003 mg · g- 1, the results were similar to those of HPLC, mostly within ± 5% .Conclusion: The method is accurate and reproducible, and provides a useful technical supplement for HPLC determination.