高效液相色谱-荧光光谱法测定大鼠血浆中葛根素及其药动学研究

目的建立高效液相色谱-荧光光谱法测定大鼠血浆中葛根素含量。方法色谱柱选用Hypersil ODS柱,流动相为甲醇∶水(含50 mmol/L醋酸铵)=23∶77(V/V),流速为1 mL/min,荧光检测器参数:激发波长250 nm,发射波长480 nm,增益为11,柱温30℃。结果该方法专属性强,在0.16~120 mg/L浓度范围内线性关系良好,最低检测限为10μg/L,准确度介于99%~104%,日间、日内精密度均小于10%。该方法成功应用于大鼠静脉注射葛根素的药动学研究中,葛根素在大鼠体内符合开放二房室模型,主要的药动学参数如下:AUC0→t(41.94±12.90)mg/(L.h),AUC0→∞(44.37±28.90)mg/(L.h),MRT(0.97±0.37)h,T1/2(1.06±0.39)h,Vss(0.09±0.02)L,Vz(0.14±0.03)L,Cl(0.10±0.05)L/h。结论本方法无需柱后修饰,简单、可靠,适于大鼠血浆中葛根素的检测。

Determination of Puerarin in rat plasma by high performance liquid chromatography with fluorescence detection and its application to pharmacokinetic studies

Objective It is to develop a method of high performance liquid chromatography (HPLC) with fluorescence detection to detect the level of puerarin in rat plasma. Methods Chromatography was performed on a Hypersil ODS column using a mobile phase consisting of methanol-50 mmol/L ammonium acetate in water (23:77, v/v) with a flow rate of 1 mL/min. The fluorescence detector was set at excitation wavelength of 250 nm, emission wavelength of 480 nm and PMT-gain of 11. The column temperature was set at 30 ℃. Results The method was selective and sensitive with a detection limit of 10 μg/L. A good linear response was observed over a wide range (0.16～120.00 mg/L) in rat plasma. This validated method was applied successfully to a pharmacokinetic study in rat plasma after intravenous administration of puerarin. The main pharmacokinetic parameters were as follows: AUC0→t (41.94±12.90) mg/(L/h), AUC0→∞(44.37±28.90)mg/(L/h), MRT (0.97±0.37) h, T1/2α (0.13±0.02) h, T1/2β (1.06±0.39) h, Vss (0.09±0.02) L, Vz (0.14±0.03) L, Cl (0.10±0.05) L/h. Conclusion A simple, rapid, sensitive and selective method of HPLC with fluorescence detection has been developed and validated for quantitative determination of puerarin in rat plasma.