LC-MS法测定人血浆中的甘草次酸

LC-MS determination of glycyrrhetic acid in human plasma

目的:建立测定人血浆中甘草次酸的 LC-MS 方法。方法:人空白血浆0.5 mL 中加入甘草次酸及内标格列喹酮,以乙酸乙酯萃取后取上清液挥干,再用流动相溶解,进行 LC/MS 测定。色谱条件:岛津 VP-ODS 色谱柱(2 μm,150 mm×2.0mm),甲醇-3 mmol·L^(-1)醋酸铵水溶液-冰醋酸(92:8:2)为流动相,流速:0.2 mL·min^(-1);质谱条件:电喷雾离子化(ESI)方式,采用选择性离子检测(SIM),检测离子为正离子,甘草次酸 SIM 的离子为[M+H]^+(m/z 471),内标格列喹酮 SIM 的离子为[M+H]^+(m/z 528)。结果:本方法线性范围为5～500 ng·mL^(-1),最低定量限(LOQ)为5 ng·mL^(-1)。准确度、精密度以及稳定性均符合有关要求。结论:本法简便,灵敏度高,可用于药代动力学试验中人血浆的甘草次酸浓度测定。

Objective：To develop a liquid chromatography -mass spectrometry（ LC -MS） method for the determination of glyeyrrhetie acid in human plasma. Methods： Gliquidone （ internal standard） and glyeyrrhetie acid was added to 0. 5 mL plasma. Then glyeyrrhetie acid and gliquidone were extracted from plasma with ethyl acetate, after transferred and evaporated, the residue was dissolved with mobile phase and determined by LC - MS method. Separation of the analytes was achieved on a Shimadzu VP- ODS analytical column（2μm,150 mm ×2. 0 mm） with a mobile phase of methanol-3 mmol·L^-1 ammonium acetate water solution-glacial acetic acid （92： 8： 2）at a flow rate of 0. 2 mL ·L^-1. MS determination in eleetrospray ionization（ESI） positive mode and in the selected ion monitoring mode（SIM）. For glyeyrrhetie acid monitored at m/z 471 and gliquidone at 528. Results ：The assay had a calibration range from 5 to 500 ng ·L^-1and a lower limit of quantification（LOQ） of 5 ng ·L^-1 for glyeyrrhetie acid. The accuracy, precision （ intra - day and inter - day）, sensitivity, specificity and stability （ ambient temperature,freeze/thaw,freeze storage） of the method fulfilled the guideline of the analytical validation criteria. Conclu- sion：This method is simple and sensitive, and suitable for the determination of glyeyrrhetie acid in human plasma.