高效液相色谱法测定人血浆中那格列奈的含量

HPLC Determination of Nateglinide in Human Plasma

目的:建立高效液相色谱法测定人血浆中那格列奈含量。方法:使用固相C18提取柱,将那格列奈和双氯芬酸钠(内标)从血浆中提取,在ODS 5μm,4.6mm×150mm柱上,以流动相0.05 mol·L-1磷酸二氢钾溶液-乙腈(16:10;用1 mol·L-1氢氧化钠溶液调节pH 6.3)进行分离,流速1 mL·min-1,210nm波长检测。结果:血药浓度线性范围0.05-20μg·mL-1(r=0.9996),血浆中最低检测浓度0.05 μg·mL-1,萃取回收率在82.4%-92.2%,相对回收率98.4%-99.7%,日内及日间RSD分别为4.3%-8.2%和4.0%-8.6%。结论:本法简便、准确,精密度高,重现性好,可应用于那格列奈人体药代动力学研究和生物等效性研究。

Objective: To establish HPLC method of the determination of nateglinide in human plasma. Method: The drug and diclofenac sodium (internal standard) were extracted from plasma using a reversed phase C18 extraction column(Sep - pak C18). Separation was accomplished on an ODS reversed phase column(5μm,4. 6 mm×150 mm) with a mobile phase of 0. 05 mol·L-1 potassium dihydrogen phosphate - acetonitrile(16: 10;adjusted pH to 6. 3 with 1 mol·L-1sodium hydroxide solution). The flow rate was 1 mL·min-1. Quantitation was a chieved by monitoring the ultraviolet absorbance at 210 nm. Results:The linear range of calibration curve was within drug plasma concentration of 0. 05 -20μg·mL-1 (r=0.9996 ) , the detection limit concentration was 0. 05μg·mL-1 in human plasma,the extraction recoveries were 82. 4% - 92. 2%. the relative recoveries were 98. 4% -99. 7% . The within - day RSD and between - day RSD were 4. 3% - 8. 2% and 4. 0% - 8. 6% respectively. Conclusion: The method possesses merits of simplicity, good accuracy, sensitivity, high precision and fine reproducibility. The method has been successfully used for analysis of plasma samples form human following oral administration of nateglinide.