新型抗帕金森氏病化合物FLZ与人和大鼠血浆蛋白结合率的测定

 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。

Determination of Human and Rat Plasma Protein Binding Rate of the Novel Anti-Parkinson’s Disease Candidate Drug FLZ

OBJECTIVE To establish an HPLC method for the determination of compound FLZ, a novel synthetic analogue of natural squamosamide, in human and rat plasma and dialysate to determine the human and rat plasma protein binding rate of FLZ. METHODS Bicyclol as the internal standard, the sample was analyzed with a mobile phase consisting of methanol and water (60∶40) at a flow rate of 1.0 mL·min-1, with the detection wavelength set at 320 nm. Equilibrium dialysis combined with HPLC method was used to determine the binding rate of FLZ to human and rat plasma protein in five concentrations. RESULTS The calibration curve was linear within the concentration range from 500 to 8 000 ng·mL-1 in human and rat plasma and from 25 to 500 ng·mL-1 in dialysate, the recovery of quality control sample with three concentration was all more than 90.3% in human and rat plasma, and more than 91.7% in dialysate, the relative standard deviation (RSD)for intra-day precision was less than 4.1% and less than 6.2% for inter-day precision in both plasma and dialysate. The average human and rat plasma protein binding rate of FLZ were 92.3% and 94.1%, respectively. CONCLUSION The established HPLC method was validated to be a simple, rapid and reliable procedure in human and rat plasma and dialysate, FLZ shows high plasma protein binding rate and there was no species difference between human and rat.