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目的建立测定人和大鼠血浆及透析外液中番荔枝酰胺衍化物(FLZ)浓度的高效液相色谱(HPLC)方法,测定FLZ与人的血浆蛋白结合率及FLZ与大鼠的血浆蛋白结合率。方法以双环醇为内标,流动相为甲醇-水(60∶40),流速1.0 mL.min-1,检测波长为320 nm。采用平衡透析法结合HPLC法,对5个浓度的FLZ与健康人及大鼠的血浆蛋白结合率进行测定。结果人和大鼠血浆(透析内液)中FLZ在500~8 000 ng.mL-1范围内、透析外液中FLZ在25~500 ng.mL-1范围内线性关系良好;低、中、高3个浓度透析内液中FLZ的回收率均大于90.3%,透析外液中FLZ的回收率均大于91.7%。透析内液及透析外液中日内精密度RSD均小于4.1%,日间精密度均小于6.2%。5个浓度的FLZ与人和大鼠的平均血浆蛋白结合率分别为92.3%和94.1%。结论本实验建立的人和大鼠血浆及透析外液中FLZ的定量分析方法快速、简便、可靠;FLZ与人和大鼠的血浆蛋白结合率均较高,且二者无种属差异。
OBJECTIVE To establish a high performance liquid chromatography (HPLC) method for the determination of spinosad (DEX) in human and rat plasma and dialysis fluid and determine the binding rate of FLZ to human plasma protein and the binding of FLZ to plasma protein of rats rate. Methods Bicyclol was used as internal standard. The mobile phase consisted of methanol-water (60:40), flow rate 1.0 mL · min-1 and detection wavelength 320 nm. Equilibrium dialysis method and HPLC method were used to determine the plasma protein binding rate of five concentrations of FLZ to healthy human and rat. Results FLZ of human and rat plasma (dialysis solution) was in the range of 500-8 000 ng.mL-1 and the FLZ of dialysis solution was good in the range of 25-500 ng.mL-1. The recoveries of FLZ in the three dialysate solutions were all over 90.3%, and the recoveries of FLZ in the dialysate outside solution were all above 91.7%. The intra-day precision RSD of dialysis solution and dialysis solution were less than 4.1%, and the intra-day precision was less than 6.2%. The mean plasma protein binding rates of FLZ at 5 concentrations to humans and rats were 92.3% and 94.1%, respectively. Conclusion The method of quantitative analysis of FLZ in plasma and dialysis extracorporeal fluid of human and rat established in this experiment is rapid, simple and reliable. The plasma protein binding rate of FLZ to human and rat is high, and there is no species difference between them.