LC-MS/MS测定大鼠血浆中K15的浓度及其在药动学研究中的应用

Determination of K15 Concentration in Rat Plasma by LC-MS/MS and Its Application in the Pharmacokinetics Study

关键词:  (3AS,4S,6AR)-四氢-4-甲氧基-呋喃并[3,4-B]呋喃-2(3H)-酮  高效液相色谱串联质谱法  药动学  SD大鼠

Key words:  (3AS, 4S, 6AR)-4H,4-methoxy furan[3,4-B] furan-2(3H)-ketone  LC-MS/MS  pharmacokinetics  SD rats

摘要:

目的 建立SD大鼠血浆中（3AS，4S，6AR）-四氢-4-甲氧基-呋喃并[3，4-B]呋喃-2（3H）-酮（K15）的LC-MS/MS测定方法，并进行K15的药动学研究。方法 分别灌胃及静脉注射给予大鼠不同剂量K15后，从眼眶静脉丛取血，并采用LC-MS/MS测定大鼠血浆中K15的浓度变化。利用DAS药动学软件拟合主要药动学参数AUC、C_max、T_max、T_1/2等。结果 大鼠血浆中的内源性杂质不干扰K15的测定，日内精密度4.53%~6.60%，日间精密度5.19%~8.14%，准确度为96.10%~102.49%。K15的线性范围为25~1 000 ng·mL^-1，r=0.998 5。K15的定量下限为25 ng·mL^-1（RSD=12.7%，n=6）。150，450和1 000 mg·kg^-1灌胃给药的生物利用度分别为79.5%，44.4%和57.0%。结论 该方法适用于K15在大鼠中的药动学研究。K15在大鼠中的药动学为非线性动力学，灌胃给予大鼠后在其体内具有一定的系统暴露量，但没有随给药剂量呈线性增加。在给药剂量为100和450 mg·kg^-1时，其在体内的C_max没有显著性增高，但是在1 000 mg·kg^-1时，C_max显著增加。随着给药剂量的增加，K15在大鼠体内的T_1/2显著增加，提示K15在大鼠体内的暴露时间随剂量的增加显著延长。

Abstract:

OBJECTIVE To establish a LC-MS/MS method for the determination of (3AS, 4S, 6AR)-4H,4-methoxy furan[3,4-B] furan-2(3H)-ketone(K15) in rat plasma and for the pharmacokinetic study of K15. METHODS After different doses of K15 were administered by intragastric or intravenous injection, blood was taken from the orbital venous plexus of rats, and the concentration of K15 in rat plasma was determined by LC-MS/MS. DAS software was used to fit pharmacokinetics parameters such as AUC, C_max, T_max, T_1/2, and so on. RESULTS The endogenous impurities in plasma did not interfere with the determination of K15. The intra-day precision was ranged in 4.53%-6.60%, the inter-day precision was ranged in 5.19%-8.14%, and the accuracy was ranged in 96.10%-102.49%. The linear range of K15 was 25-1 000 ng·mL^-1, r=0.998 5. The lower limit of quantitation of K15 was 25 ng·mL^-1(RSD=12.7%, n=6). The bioavailability were 79.5%, 44.4% and 57.0% for administrated at the dose of 150, 450 and 1 000 mg·kg^-1, respectively. CONCLUSION The method is suitable for the analysis of K15 in rat plasma. The pharmacokinetics of K15 in rats is nonlinear dynamics. K15 has a certain amount of systemic exposure in rats after orally administrated to rats, but the systemic exposure dose not increase linearly with the amount of treated. When K15 is administered at 100 and 450 mg·kg^-1, there is no significant increase in C_max in vivo, but C_max increases significantly at 1 000 mg·kg^-1. With the increase of dosage, the T_1/2 of K15 increases significantly in rats, suggesting that the exposure time in rats increase significantly with the increase of dosage.