大鼠血浆中山奈酚的HPLC测定法及其在药物动力学研究中的应用

目的建立测定大鼠血浆中山奈酚质量浓度的HPLC法,并用于山奈酚经灌胃给药后在大鼠体内药动学研究。方法大鼠灌胃给予200 mg.kg-1山奈酚混悬液,于给药后不同时间采集血样,以非那西丁为内标,VC为抗氧剂,采用2 mol.L-1盐酸,80℃水浴水解30 min后用乙醚萃取的方法处理血浆样品。采用RP-HPLC法测定山奈酚血药质量浓度,色谱柱为Diamonsil C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-体积分数为0.05%磷酸溶液(体积比40∶60),流速为1.0 mL.min-1,检测波长为370 nm,柱温为40℃。结果血浆中内源性物质对山奈酚测定无干扰,线性范围为0.050～10 mg.L-1,r=0.998 5,回收率、准确度及日内、日间精密度均符合生物样品测定要求,定量下限(LLOQ)为50μg.L-1。山奈酚主要药动学参数为AUC0-t=129.2 mg.h.L-1,ρmax=7.39mg.L-1,tmax=9.0 h,t1/2=8.3 h。结论该方法简便、快速、重复性好,适用于山奈酚大鼠血药质量浓度测定及体内药动学研究。

Determination of kaempferol in rat serum by HPLC and its application in pharmacokinetic study

Objective To establish an HPLC method for determining plasma level of kaempferol and investigate its pharmacokinetics in rats.Methods Rats were orally administrated with kaempferol suspension(200 mg · kg-1)and then blood samples were taken.The internal standard was phenacetin,VC was added as antioxidant.The sample was spiked with the same volume of hydrochloric acid(2 mol · L-1)and hydrolyzed for 30 min in a water bath at 80 ℃,then followed by liquid-liquid extraction.The plasma drug levels were determined by RP-HPLC,the analysis was performed on a Diamonsil C18 column(250 mm×4.6 mm,5 μm)at 40 ℃ with acetonitrile-0.05% phosphoric acid(V∶ V=40∶ 60)as the mobile phase at the flow rate of 1.0 mL · min-1.The detecting wavelength was set at 370 nm.Results The linearity was confirmed in the concentration range of 0.050-10 mg · L-1 with the correlation coefficient of 0.998 5.The limit detection(LLOQ)was 50 μg · L-1.The variability and relative recovery rate were suitable for biological analysis.The main pharmacokinetic parameters were calculated as follows:AUC0-t=129.2 mg · h · L-1,ρmax=7.39 mg · L-1,tmax=9 h,t1/2=8.3 h.Conclusions The method is convenient,rapid,accurate and can be used for pharmacokinetic study of kaempferol.