RP-HPLC法测定人血浆中扎来普隆浓度

Determination of Zaleplon in Human Plasma by RP - HPLC with Fluorescence Detection

目的:建立测定人血浆中扎来普隆浓度的反相高效液相色谱法。方法:血浆样品中加入内标β-萘酚后用乙酸乙酯提取。固定相为Supelcosil C8色谱柱(250 mm ×4.6 mm,5μm),流动相为0.04 mol·L-1磷酸二氢钾缓冲液-乙腈(6:5,含三乙胺0.3%,磷酸调至pH 5.0),流速为1.0 mL·min-1。荧光检测:荧光激发波长345nm,发射波长460 nm。结果:本方法线性范围为1-128 ng·mL-1(r=0.9995),最低检测浓度为0.2 ng·mL-1,提取回收率为87.4%-91.9%,方法回收率为98.8%-100.8%,日内RSD为2.9%-5.4%,日间RSD为3.8%-13.3%。结论:本法简便、准确,适用于扎来普隆药代动力学的研究。

Objective: An RP - HPLC method was established for the determination of zaleplon in human plasma. Methods: After addition of β- naphthol (internal standard ) , zaleplon in plasma samples was extracted with ethyl acetate and determined by RP - HPLC. The analysis involved a 250 mm×4. 6 mm column packed with Supelcosil C8 (5 μm). The mobile phase consisted of 0. 04 mol·L-1 potassium dihydrogen phosphate solution - acetonitrile (6 :5,contained triethylamine 0. 3% ,adjusted to pH 5. 0 with phosphoric acid) with a flow rate of 1. 0 mL·min-1. The fluorescence detector was set at Ex 345 nm and Em 460 nm. Results:The linear range was of 1 - 128 ng·mL-1 ( r= 0. 9995 ). The minimal detectable concentration was 0. 2 ng·mL-1. The extraction recovery was 87. 4% -91. 9%. The method recovery was 98. 8% - 100. 8% . Within - day RSD was 2. 9% -5. 4% . Between - day RSDwas 3. 8% - 13. 3%. Conclusion: The method is simple and accurate, which is suitable for the pharmacokinetic study of zaleplon.