RP-HPLC内标法测定血浆中沙利度胺的浓度

DETERMINATION OF THALIDOMIDE IN HUMAN PLASMA BY RP-HPLC INTERNAL STANDARD METHOD

目的建立高效液相色谱法测定人血浆中沙利度胺的浓度。方法采用液-液萃取方法,用乙酸乙酯提取血浆中沙利度胺,选用非那西丁为内标,色谱柱为Waters Atlantis T3 C18(4.6 mm×150 mm,5μm)反相柱,流动相:乙腈-25 mmol/L KH2PO4缓冲液(PH3.5)(25∶75);沙利度胺的检测波长为220 nm,非那西丁的检测波长为248 nm;流速为1 ml/min。结果本方法的线性范围为0.05～20μg/ml,r2=0.999 0,沙利度胺保留时间7.8 min,平均方法回收率为(95.3±8.2)%(n=15),日内RSD≤4.5%,日间≤7.8%。准确度在88.1%～95.6%之间。沙利度胺的最低检测浓度为0.05μg/ml。结论本方法灵敏、准确、选择性强,适用于临床血药浓度监测和人体药代动力学研究。

Objective To establish a HPLC method for the determination of thalidomide in human plasma. Methods A liquid liquid extraction method was used. Thalidomide in plasma was extracted with ethyl acetate, and phenacitin was used as the internal standard. The sample was separated on a Waters Atlantis T3 C18 （4. 6 mm x 150 mm ,5 μm）column, with a mobile phase of 25 mmol/L potassium dihydrogenphosphate buffer （PH3.5） acetonitrile （75：25）. Thalidomide was detected at UV wavelength of 220 nm, and phenacitin was detected at 248 nm. The flow rate was lmL/min and the column temperature was room temperature. Results The method had a chromatographic running time of 7.8 min and linear calibration curves over the thalidoide concentrations ranges of 0. 05 20 I.Lg/ml （ r2 = 0. 999 0）. The recovery of assay was （95.3± 8.2） % （ n = 15 ）. Inter day and intra day variation coefficient was less than 4. 5% and 7.8%, repectively. The detection limit of thalidomide was 0. 05μg/ml. Conclusion The method is sensitive, accurate, and selective for clinical monitoring of thalidomide and its pharmacokinetic studies.