高效液相色谱法测定大鼠血浆和子宫中黄体酮及其代谢物的浓度

目的建立高效液相色谱法测定大鼠血浆和子宫样品中的黄体酮及其代谢物20α-羟基黄体酮,并研究大鼠肌肉注射黄体酮后血浆和子宫中的药物代谢动力学。方法样品经液-液萃取后,以乙腈-水(60∶40,pH 4.0)为流动相,用ODS柱进行分离,240 nm检测。以18-甲基炔诺酮为内标。结果血浆中黄体酮C_max为(508±62) μg·L^-1,T_max为(3.2±0.4) h,T_1/2(k_e)为(10±4) h,AUC_0-48h为(5 886±1 573) μg·L^-1·h,子宫中黄体酮T_max为(5.2±1.1) h,C_max(1.7±1.1) μg·g^-1。20α-羟基黄体酮具有与黄体酮相似的T_max。结论该方法简便、准确,可同时测定黄体酮和代谢物,适用于黄体酮及其代谢物20α-羟基黄体酮的药代动力学研究。

Determination of progesterone and its main metabolite in rat plasma and uterus using HPLC

AimTo quantify progesterone(P) and one of its metabolites 20α-hydroxy-4-pregnen-3-one (20α-OHP) in rat plasma and uterus after im administration of progesterone. MethodsPlasma and uterus samples were prepared by liquid-liquid extraction and separated through Shimadzu VP-ODS column (150 mm×4.6 mm ID, 5 μm). The mobile phase consisted of acetonitrile and water (60∶40, adjusted to pH 4.0 with phosphoric acid). The detector was set at 240 nm. Norgestrel was used as the internal standard. ResultsC_max of P in plasma was (508±62) μg·L^-1, T_max was (3.2±0.4) h, T_1/2(k_e) was (10±4) h and mean AUC_0-48h was (5 886±1 573) μg·L^-1·h. The maximum concentration of P in uterus was(1.7±1.1) μg·g^-1 and the peak time was (5.2±1.1) h. 20α-OHP showed a similar T_max with P. ConclusionThe method is accurate and convenient. It can be used to determine P and its main metabolite 20α-OHP simultaneously for studying their preclinical pharmacokinetics.