ELISA、HPLC测定清燥救肺汤中甘草酸含量的比较研究.docVIP

  ELISA、HPLC 测定清燥救肺汤中甘草酸含 量的比较研究# 吴振起1，王思源1，平静1，于艳2，南春红1* 5 10 15 20 25 30 35 40 （1. 辽宁中医药大学附属医院儿科，沈阳 110032； 2. 辽宁中医药大学附属医院中药实验室，沈阳 110032） 摘要：目的：建立免疫分析技术对清燥救肺汤中甘草酸(GA)的含量分析方法。方法：采用 抗 GA 的单克隆抗体（GA-MAb）进行竞争性酶联免疫吸附（ELISA）法测定清燥救肺汤中 GA 的含量，同时结合 HPLC 色谱分析方法，进行比较确证。结果：ELISA 测定清燥救肺汤 中成分 GA 在 0.3125～10 μg（r=0.9969）范围内线性关系良好，平均回收率分别为 109.0%、 92.5%和 103.9%，三批样品中 GA 含量均值为 6.3003 mg· g-1；与 HPLC 测定结果相近，RSD ＜5％。结论：ELISA 对中药成分测定准确可靠，重复性好，是 HPLC 测定有益的技术补充。 关键词：清燥救肺汤；甘草酸；ELISA；HPLC；含量测定 ELISA and HPLC in the Determination of glycyrrhizic acid in Qingzaojiufei Tang WU Zhenqi1, WANG Siyuan1, PING Jing1, YU Yan2, NAN Chunhong1 (1. Department of Pediatrics, Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, ShenYang 110032; 2. Laboratory of Chinese Medicine, Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, ShenYang 110032) Abstract: Objective: To establish an immunoassay method for the determination of glycyrrhizic acid(GA) in Qingzaojiufei Tang. Methods: An enzyme linked immunosorbent assay(ELISA) using monoclone antibody to determine the content of GA, combining with the analysis method of HPLC to confirm. Results: GA had good linearity in the ranges of 0.3125～10 μg (r=0.9969), the average recoveries of GA were 109.0%, 92.5% and 103.9%, respectively. The average content of GA in three sample was 6.3003 mg·g-1 by using ELISA. The RSD of GA determinations between ELISA and HPLC less than±5％，showed a high similarity. Conclusions: The developed ELISA method is accurate with high repeatability, which is helpful to provides useful technical supplement for the determination of HPLC. Keywords: Qingzaojiufei Tang; glycyrrhizic acid; ELISA; HPLC; content determination 清燥救肺汤是治疗燥热伤肺的经典名方，首见于《医门法律》，为清代医家喻嘉言创制， 由桑叶、石膏、麦冬、杏仁、甘草、人参、阿胶、枇杷叶、胡麻仁组成。此方重用桑叶为君 药，轻清宣泄肺中燥热；杏仁、石膏、枇杷叶宣中有清，清中有降，宣肺气、清肺热而肃降 有权；人参、甘草、胡麻仁、阿胶、麦冬益气养阴，培土生金，补肺润肺以扶正[1]。清燥救 肺汤在临床上广泛应用，然而目前尚缺乏质量控制方面的研究。现代研究表明，甘草酸(GA) 具有抗肿瘤、抗炎、抗病毒、增强免疫功能等作用[2]，可作为甘草及其制剂的定量指标，用 于评价制剂的稳定性、优劣制订药品质量标准等。