应用微乳液相色谱法测定大鼠脑脊液中槲皮素-3-O-槐糖苷含量研究

Determination of quercetin-3-O-sophoroside in rat cerebrospinal fluid by microemulsion liquid chromatography

目的:建立测定大鼠脑脊液中槲皮素-3-O-槐糖苷含量微乳液相色谱方法,并将其用于大鼠体内药代动力学的研究。方法:采用微乳液相色谱法分别测定给药后不同时间点大鼠脑脊液中的药物浓度,并研究其药代动力学参数。结果:最佳微乳流动相组成为:2.0%Genapol X-080-2.5%正丁醇-1.5%乙酸乙酯-0.3%三乙胺-93.7%水溶液(p H 5.0)。大鼠脑脊液中槲皮素-3-O-槐糖苷在浓度0.1~10.0μg·m L^(-1)内线性关系良好(r=0.998 4),最低定量浓度(S/N=10)为0.1μg·m L^(-1),日内和日间RSD均小于4.9%,平均加样回收率为96.3%,RSD为5.1%。槲皮素-3-O-槐糖苷脑内药代动力学模型符合二室模型。分布和消除相半衰期分别为7.3 min和28.7 min。药-时曲线下面积(AUC)为804.37μg·min^(-1)·m L^(-1)。表观分布容积(V_d)为0.325μg·m L^(-1)。平均驻留时间(MRT)为25.8 min。结论:本微乳液相色谱法已成功的用于测定大鼠脑脊液中槲皮素-3-O-槐糖苷含量的测定及药代动力学研究。

Objective: To establish a microemulsion liquid chromatography method for the determination of quercetin-3-O-sophoroside in rat cerebrospinal fluid, and further to apply this method to pharmacokinetics study of quercetin-3-O-sophoroside in rats. Methods: After intravenous injection of quercetin-3-O-sophoroside, the cerebrospinal fluid concentrations of rats at different time points were determined by microemulsion liquid chromatography. At the same time, the pharmacokinetic parameters were studied. Results: The optimized microemulsion mobile phase consisted of 2.0% (v/v) of Genapol X-080, 2.5% (v/v) n-butyl alcohol, 1.5% (v/v) ethyl acetate, 0.3% (v/v) triethylamine and 93.7% (w/v) of aqueous solution (pH = 5.0). The calibration curve for quercetin-3-O-sophoroside was linear in the range of 0.1 – 10.0 μg·mL-1 (r = 0.998 4). The intra-day and inter-day precision (RSD) were below 4.9% and the limits of detection (LOD) was 0.1 μg·mL-1 (S/N = 10). The average rate of recovery was 96.3% and RSD was 5.1%. The pharmacokinetic course of quercetin-3-O-sophoroside in rats fitted the two-compartment model. The relavent pharmacokinetic parameters were as follows: t1/2α = 7.3 min, t1/2β = 28.7 min, AUC = 804.37 μg·min-1·mL-1, Vd = 0.325 μg·mL-1, MRT =25.8 min. Conclusion: The microemulsion liquid chromatography method was successfully applied to determine the concentration of quercetin-3-O-sophoroside in rat cerebrospinal fluid and to the pharmacokinetics research in rats.