HPLC测定血浆和尿液中那格列奈对映体浓度

Determination of Nateglinide Enantiomer in Human Plasma and Urine by HPLC

目的:建立手性色谱柱拆分血浆和尿液中那格列奈对映体（D-那格列奈和L-那格列奈）的简便方法。方法:采用Chiralcel OD-R （10μm,0.46cm×25cm）手性色谱柱,流动相为乙腈-0.5mol·L-1高氯酸钠（70:30）,0.5mol·L-1高氯酸调pH2.2,检测波长214nm,流速:0.4mL·min-1,内标法室温测定。结果:血浆中D-那格列奈的线性范围是0.02～20mg·L-1,r=0.9995;L-那格列奈的线性范围是0.08～20mg·L-1,r=0.999 4 ;尿液中D-那格列奈的线性范围是0.02～10mg·L-1,r=09998;L-那格列奈的线性范围是0.08～10mg·L-1,r=0.9997;血浆中D-那格列奈高中低3个质控样品的日内RSD≤6.9％,日间RSD≤8.2％,L-那格列奈高中低3个质控样品的日内RSD≤7.1％,日间RSD≤10.0％;尿液中D-那格列奈高中低3个质控样品的日内RSD≤7.0％,日间RSD≤9.8％,L-那格列奈高中低3个质控样品的日内RSD≤7.3 ％,日间RSD≤10.3％。结论:本方法分离度好,简便快速,完全适用于D-那格列奈人体药代动力学及肝代谢过程中消旋化的研究。

Objective: To establish a simple method for the determination of nateglinide enantiomers in human plasma and urine. Method: HPLC was applied for quantitative analysis. Chiralcel OD - R (10μm, 0.46 cm ×25 cm) column was used. The mobile phase was consisted of acetonitrile - 0.5 mol·L-1 sodium perchlorate (pH 2.2) (70:30), and the flow rate was 0.4 mL·min-1 . The UV detection wavelength was 214 nm and the whole operation was under room temperature . Results : The linearity was obtained over the range of 0.02-2 0 mg·L-1 for D - nateglinide ( r = 0.999 5) and 0.08-20mg·L-1 for L - nateglinide ( r = 0.999 4) in plasma, respectively. The linearity was obtained over the range of 0.02-10 mg·L-1 for D - nateglinide ( r = 0.999 8) and 0.08 -10 mg·L-1 for L -nateglinide ( r = 0.999 7 ) in urine, respectively . The intra - day and inter - day RSD for D - nateglinide in plasma were less than 6.9% and 8.2% ( n = 5 ) , respectively. The intra - day and inter - day RSD for L - nateglinide in plasma were less than 7.1% and 10.0% ( n = 5 ), respectively. The intra- day and inter- day RSD for D -nateglinide in urine were less than 7.0% and 9.8% ( n = 5 ) , respectively. The intra- day and inter- day RSD for L - nateglinide in urine were less than 7.3% and 10.3% (n=5), respectively. Conclusion: The assay was rapid and simple to allow accurate and precise measurements of D - nateglinide and its enantiomer in plasma during pharma-cokinetic studies in human.