HPLC法同时测定小鼠血浆中3种银杏酸单体的浓度

目的: 建立小鼠血浆中银杏酸（ginkgolic acid，GA) 3种主要单体GA 13∶0，GA 15∶1和GA 17∶1的高效液相色谱(HPLC)测定方法。方法: 以制备的GA 13∶0，GA 15∶1和GA 17∶1单体为标准品，采用外标法建立标准曲线。采用乙腈预处理样品以沉淀其中蛋白质。采用SinoChrom ODS AP C18色谱柱 (250 mm×4.6 mm，5 μm)，甲醇-3%醋酸(92∶8)为流动相，流速1.0 mL· min^-1，检测波长为310 nm。结果： GA 13∶0，GA 15∶1和GA 17∶1标准曲线分别在0.135～20μg·mL^-1，0.129 ～20μg·mL^-1和0.211～22μg·mL^-1范围内线性关系良好（r均＞0.999 0）；回收率均＞70.2% (n=5)，测定准确度为95.2%~101.5%；日内相对标准偏差≤6.2%，日间相对标准偏差≤9.5%。结论：成功建立测定GA 13∶0，GA 15∶1和GA 17∶1单体的HPLC方法，其灵敏度高，重现性、精密度和准确性好。

High performance liquid chromatography determination method for three monomers of ginkgolic acid in mouse plasma

Objective: To develop a high performance liquid chromatography (HPLC) method for determining three monomers of ginkgolic acids, GA 13∶0, GA 15∶1 and GA 17∶1,  in mouse plasma. Methods: Plasma samples were treated with acetonitrile for protein precipitation. The analytical column was SinoChrom ODS AP C18(250 mm×4.6 mm, 5 μm).The mobile phase consisted of a mixture of methanol 3% acetic acid (92∶8) at a flow rate of 1.0 mL· min^-1. The detection wavelength was at 310 nm. Results: Standard curves of GA 13∶0, GA 15∶1 and GA 17∶1 were linear in the range  of 0.135-20 μg·mL^-1, 0.129-20 μg·mL^-1, and 0.211-22 μg·mL^-1, respectively (both r＞0.9990). The lower limit of quantification were 0.135, 0.129 and 0.211 μg·mL^-1, respectively. The extraction recoveries＞70.2% (n=5) and the accuracies were 95.2%-101.5%. The intra-day and inter-day precisions were lower than 6.2% and 9.5 % (n=5), respectively. Conclusion: The HPLC method was successfully built for determining GA 13∶0, GA 15∶1 and GA 17∶1, which was simple and  sensitive, accurate with  reproducible, and suitable for clinical pharmacokinetic study.  ［Key words］ginkgolic acid monomer; high performance liquid chromatography; mouse