HPLC法测定小鼠血浆中洋川芎内酯Ⅰ的浓度及药动学研究

目的:建立测定小鼠血浆中洋川芎内酯Ⅰ浓度的高效液相色谱方法,并应用于药动学研究。 方法:小鼠单剂量静注或灌胃给予(104 mg·kg^-1)洋川芎内酯Ⅰ,在给药后不同时间取血,血浆经沉降蛋白处理后,采用HPLC法测定洋川芎内酯Ⅰ的浓度,使用Megres-C₁₈色谱柱(150 mm×4.6 mm,5 μm),以乙腈-2.0%冰醋酸(1: 3,v/v)为流动相,流速为1.0 mL·min^-1,柱温为30℃,洋川芎内酯H为内标,检测波长278 nm。 结果:在0.075~30 mg·L^-1范围内,洋川芎内酯Ⅰ在血浆中线性关系良好,定量下限为0.075 mg·L^-1,方法回收率大于85%,日内日间 RSD 均小于5%,样品在室温下放置24 h,-4℃放置5 d,经过3次冻融循环后基本稳定。小鼠静注给药后主要药代动力学参数分别为t_1/2 31.93 min,AUC_(0-∞) 2738.80 mg·min·L^-1,MRT_(0-∞) 31.61 min;小鼠灌胃给药后主要药代动力学参数分别为t_1/2 40.07 min,AUC_(0-∞) 892.42 mg·min·L^-1,MRT_(0-∞) 61.38 min。经计算,洋川芎内酯Ⅰ在小鼠体内的绝对生物利用度为32.19%。 结论:所建立的方法可靠,简便快速,适用于洋川芎内酯Ⅰ小鼠体内血药浓度测定及药代动力学的研究。

Objective: To establish an HPLC method for the determination concentration of senkyunolideⅠin mice plasma for pharmacokinetic study. Methods: The mice were intravenously administrated of a single dose or intragastricly administrated senkyunolide I(104 mg·kg^-1).The plasma samples after protein settlement processing were analyzed for the senkyunolideⅠ concentration by HPLC.The Megres-C₁₈(150 mm×4.6 mm,5 μm)column was adopted,the mobile phase was acetonitrile-0.2% glacial acetic acid(1: 3,v/v)at a flow rate of 1.0 mL·min^-1 under 30℃,the senkyunolide H was marked as the internal standard,and the detection wavelength was set at 278 nm. Results: Excellent liner relationship was obtained in the range of 0.075 to 30 mg·L^-1,the limit determination of senkyunolideⅠwas 0.075 mg·L^-1.The recovery of the method was more than 85%,and the intra-and inter-day RSDs were less than 5%.The samples were stable for 24 h at room temperature,5 days at-4℃,and remained stable after three freeze-thaw cycles.The pharmacokinetic parameters were as follows:t_1/2 31.93 min,AUC_(0-∞) 2738.80 mg·min·L^-1,MRT_(0-∞) 31.61 min in mice with intravenous administration,and t_1/2 40.07 min,AUC_(0-∞) 892.42 mg·min·L^-1,MRT_(0-∞) 61.38 min in mice with intragastric administration.The absolute bioavailability of senkyunolideⅠ in mice was calculated to be 32.19%. Conclusion: The method is reliable,simple,rapid,and suitable for the pharmacokinetic study and determination of senkyunolideⅠ in plasma.