RP-HPLC法测定大鼠血浆中DHX-11的浓度

Determination of content of DHX-11 in rat plasma by RP-HPLC

目的建立测定大鼠血浆中10-O-[4-(1-乙酰基-5-苯基-4,5-二氢吡唑-3-基)苯基]-(10S)-二氢青蒿素(10-O-[4-[1-acetyl-5-phenyl-4,5-dihydropyrazol-3-yl]phenyl]-(10S)-dihydroartemisinin,DHX-11)的高效液相色谱方法,并应用于DHX-11的药物动力学研究。方法采用反相色谱柱(200mm×4.6 mm,5μm),流动相为甲醇-水(体积比为90∶10),流速为1 mL.min-1,检测波长为294nm,内标为大黄酚,以液液萃取法处理血浆。结果 DHX-11血浆浓度在0.022～1.120 mg.L-1内线性关系良好(r>0.996 0),定量下限为0.022 mg.L-1,日内、日间精密度(以相对标准偏差表示)不大于13.3%,低、中、高质量浓度(0.0448、0.224、0.896 mg.L-1)质控样品回收率分别为92.7%、91.1%、85.7%。大鼠灌胃给药血药浓度-时间数据经DAS2.0药动学软件处理,主要药动学参数为:t1/2:(1.922±0.632)h;CL/F:(104.673±31.629)L.h-.1kg-1;AUC0-t:(3 778.6±1 031.4)μg.h.L-1;AUC0-∞:(3 879.3±1 010.7)μg.h.L-1。结论该方法可作为大鼠血浆中DHX-11浓度的测定方法,为探讨DHX-11的药动学行为提供了方法依据。

Objective To develop an HPLC-UV method for determination of 10-O-[4-[1-acetyl-5-phenyl-4,5-dihydropyrazol-3-yl]phenyl]-（10S）-dihydroartemisinin（DHX-11）in rat plasma.Methods A reverse-phase column（200 mm×4.6 mm,5 μm）was applied with mobile phase consisted of methanol-water（V∶ V=90∶ 10）at a flow rate of 1 mL · min-1,the detection wavelength was set at 294 nm,chrysophanol was used as the internal standard,and the plasma was prepared by liquid-liquid extraction.Results The standard curve was linear over the range of 0.022-1.120 mg · L-1 in rat plasma with correlation coefficient more than 0.996.The lower limit of quantification of the assay was 0.022 mg · L-1.The relative standard deviations of intra-and inter-day assay variations were less than 13.3%.The recoveries of samples at 0.045,0.224 and 0.896 mg · L-1 were 92.7%,91.1% and 85.7% respectively.The plasma concentration-time data after oral administration was calculated by DAS2.1 pharmacokinetic program,and the main pharmacokinetic parameters were：t1/2：（1.922±0.632）h;CL/F：（104.673±31.629）L · h-1 · kg-1;AUC0-t：（3 778.6±1 031.4）μg · h · L-1;AUC0-∞：（3 879.3±1 010.7）μg · h · L-1.Conclusions The method is simple,rapid and accurate and can be applied to investigate the pharmacokinetics of DHX-11 in rats.