高效液相色谱-荧光结合蛋白沉淀法测定人血浆中格列喹酮及相对生物利用度

Determination of gliqudone in human plasma by HPLC-fluorescence and bioequivalence of 2 gliqudone tablets

目的建立HPLC结合荧光测定血浆中格列喹酮的方法，研究2种格列喹酮片剂的相对生物利用度。方法以甲醇-20mmol·L^-1磷酸二氢氨（氨水调pH为7．35）=60：40（v／v）为流动相，样品沉淀蛋白后直接进样，荧光检测器的EX为315nm，EM为410nm。18名健康志愿者采用随机交叉试验设计，分别单剂量口服格列喹酮片60mg后测定两者相对生物利用度。结果血浆中格列喹酮与内源性杂质分离完全，在30．5～1950μg·L^-1浓度与峰面积线性关系良好，回归方程为：A=797．7C-3．320×10^4（r^2=0．9991），最低定量限为30．5μg·L^-1；绝对回收率为81．7％～103．6％（n=15），方法回收率为89．3％～108．7％（n=15），日内精密度（RSD）〈1．3％，日间精密度（RSD）〈2．4％，以AUC0～24为指标，试验制剂相对于参比制剂的生物利用度为（101．5±13．3）％。结论该法简单，准确度高，灵敏度好。方差分析结果表明试验制剂与参比制剂的主要药动学参数之间无明显差异，试验制剂与参比制剂为生物等效制剂。

Objective To determine gliqudone in the plasma and evaluate the bioequivalence of 2 gliqudone tablets （T and R） by HPLC-fluorescence method. Methods Gliqudone was separated on a C18 column with a mixture of methanol-phosphate buffer solution （60 ： 40, v/v, pH value was adjusted to 7.35 by NH3 · H2O） as the mobile phase, and was detected at EX： 315 nm, EM： 410 nm. A single oral dose of 60 nag gliqudone was administered to 18 healthy volunteers in a randomized crossover study, and the pharmacokinetics and bioavailability were studied. Results Gliqu- done was separated well with the endogenous substances. The calibration curve was linear at 30. 5～1 950 μg·L-1 with r^2=0. 999 1, the absolute recovery was at 81.7% ～103.6% （n=15）, and the method recovery was 89.3%～ 108.7% （n= 15）. Inter-day RSD〈1. 3% and intra-day RSD（2.4%, with the AUC0-24 as the index, the relative bioavailability of tablet T was （101.5±13. 3）% when compared with tablet R. Conclusion The established method is accurate and sensible for the determination of gliqudone in human plasma. Statistic analysis showed no significant difference between the two preparations, which are bioequivalent.