人血浆中西沙必利的高效液相色谱测定法

HPLC determination of the concentration of cisapride in human plasma

目的 :建立人血浆中西沙必利的高效液相色谱测定法。方法 :取血浆 2 0 0 μL ,碱化后用正庚烷 -异戊醇 (95∶5 )一步提取 ,常温下氮气吹干。采用Shim -packCLC -ODS色谱柱 ;流动相为磷酸二氢钾溶液 (pH =4 5 ,0 0 2mol·L- 1 ) -乙腈=5 7∶43,流速 1 3mL·min- 1 ;检测波长 315nm ;柱温 35℃。结果 :西沙必利与内标物的保留时间分别为 4 0min和 5 4min ;西沙必利浓度C在 4～ 2 0 0 μg·L- 1 范围内与西沙必利 内标物的峰高比PHR呈良好线性关系 ,回归方程为 :PHR =0 0 0 8193C +0 0 0 2 75 4,r=0 9997,n =9,,斜率RSD =4 82 %。高中低 3个质控样本的批内 (n =10 )及批间 (n =9)RSD分别为 2 91%～ 3 5 7%和 3 30 %～ 3 99% ,方法学回收率为 96 %～ 10 0 % ,最低可检出浓度为 2 μg·L- 1 。除妥拉苏林外 ,西沙必利代谢物Norcisapride及其它被测药物对西沙必利和内标物均无干扰。结论 :本方法取样量小、灵敏度高、操作便捷 ,为血药浓度监测及临床药代动力学研究提供了方法学基础。

Objective:To improve a HPLC method for the determination of cisapride in human plasma.Method:Alkalinized samples of plasma(200 μL)were extracted with 1.0 mL of 5% isopropanol in heptane.The organic phase was evaporated to dryness under nitrogen at ambient temperature.The residues was redissolved in mobile phase and injected.Cisapride and internal standard were analyzed on Shim-pack CLC-ODS column(6.0 mm×150 mm,5 μm)at 35 ℃ with a 0.02 mol·L -1,(pH 4.5)potassium dihydrogen phosphate solution-acetonitrile(57∶43,v/v)mobile phase and detected at 315 nm.The flow rate was 1.3 mL·min -1.Result:Cisapride and internal standard were eluted at 4.0 min and 5.4 min,respectively,after injection.Calibration plots in human plasma were linear(r=0.9997,n=9)from 4 to 200 μg·L -1.For 8,20,80 μg·L -1 check samples,intra-run(n=10)precisions(RSD) were 2.91%～3.57% and inter-run(n=9)precisions (RSD)were 3.30%～3.99%,respectively.The average recoveries were 100%,96% and 97% for the low,middle and high check samples,respectively.The limit of detection was 2 μg·L -1.Edogenous plasma components,the major cisapride metabolite(norcisapride)and other drugs(not including tolazoline)used in pharmacotherapeutics did not interfere.Conclusion:The improved analytical method for cisapride was found to be sensitive,simple and rapid,suitable for application in pharmacokinetic studies and routine determination of numerous samples.