Zhao Yan, Qu Huihua, Wang Xueqian, Zhang Yue, Shan Wenchao, Zhao Yan, Wang Qingguo
School of Basic Medical Sciences, Beijing University of Chinese Medicine, Beijing, China.
Scientific Research Experiment Center, Beijing University of Chinese Medicine, Beijing, China.
Planta Med. 2015 Jun;81(9):765-70. doi: 10.1055/s-0035-1545913. Epub 2015 Apr 9.
This study aimed to develop an indirect competitive enzyme-linked immunosorbent assay based on monoclonal antibodies against paeoniflorin to study the effects of different doses of glycyrrhizinic acid on the pharmacokinetics of paeoniflorin in mice. An anti-paeoniflorin monoclonal antibody was produced from a hybridoma created through a fusion of splenocytes immunized with paeoniflorin-bovine serum albumin and conjugated with the hypoxanthine-aminopterin-thymidine-sensitive mouse myeloma cell line SP2/0. The resultant antibody was used to develop and validate a rapid, specific and sensitive, indirect competitive enzyme-linked immunosorbent assay for the measurement of paeoniflorin (linear range 4.8-312.5 ng · mL(-1)). The intraday and interday precision values of the indirect competitive ELISA method were well within the recommended range (≤ 10 %), and the recovery rate was, on average, 101.13 %. Pharmacokinetic parameters obtained from mouse blood samples at various intervals following the oral administration of paeoniflorin and glycyrrhizic acid at three doses (1 : 0.3, 1 : 1, 1 : 3, respectively) demonstrated that the highest dose of glycyrrhizic acid inhibits the absorption of paeoniflorin.
本研究旨在开发一种基于抗芍药苷单克隆抗体的间接竞争酶联免疫吸附测定法，以研究不同剂量甘草酸对小鼠体内芍药苷药代动力学的影响。通过用芍药苷-牛血清白蛋白免疫的脾细胞与次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷敏感的小鼠骨髓瘤细胞系SP2/0融合产生的杂交瘤制备抗芍药苷单克隆抗体。所得抗体用于开发和验证一种快速、特异且灵敏的间接竞争酶联免疫吸附测定法，用于测定芍药苷（线性范围4.8 - 312.5 ng·mL⁻¹）。间接竞争ELISA法的日内和日间精密度值均在推荐范围内（≤10%），平均回收率为101.13%。在分别以三种剂量（1∶0.3、1∶1、1∶3）口服给予芍药苷和甘草酸后，在不同时间间隔从小鼠血样中获得的药代动力学参数表明，最高剂量的甘草酸会抑制芍药苷的吸收。
