高效液相色谱法测定人肝微粒体中帕洛诺司琼的浓度

 目的研究帕洛诺司琼的体外代谢,建立人肝微粒体中帕洛诺司琼的反相高效液相色谱测定法。方法帕洛诺司琼与人肝微粒体共孵育之后用乙醚提取,采用地非三唑为内标,以Diamonsil C₁₈柱(4.6mm×200mm,5μm)为分析柱,0.01μmol·L^-1KH₂PO₄(pH3.0)-甲醇(30∶70)为流动相,流速1.0mL·min^-1,紫外检测波长为241nm。结果帕洛诺司琼在1～100μmol·L^-1内线性关系良好(r=0.9999,n=5),检测限为0.05μmol·L^-1(S/N≥3),定量限为(0.21±0.04)μmol·L^-1(n=5)。方法提取回收率和方法回收率平均分别为89.7%和100.0%,日内,日间RSD均<10%(n=5)。结论此法简便,准确,可用于研究帕洛诺司琼在人肝微粒体中的代谢。

Determination of Palonosetron in Humnan Liver Microsomes by High-Performance Liquid Chromatography with UV-Detection

OBJECTIVE To establish a RP-HPLC method for determinating palonosetron in human liver microsomes. METHODS Palonosetron in human liver microsomal incubates was extracted by diethyl ether and diphenytriazol was used as internal standard. The determination was performed on a Diamonsil C₁₈ column(4.6 mm×200 mm, 5 μm) with a mobile phase consisting of 0.01 mol·L^-1 KH₂ PO₄ (pH 3.0)and methanol (30∶70) at a flow-rate of 1.0 mL·min^-1.UV detector was at 241 nm.RESULTS The assay was linear over the range of 1～100 μmol·L^-1 for palonosetron (r=0.999 9).The limit of detection was 0.05 μmol·L^-1(S/N≥3) and the limit of quantification was (0.21±0.04) μmol·L^-1(n=5).The assay recovery and the absolute recovery were 89.7% and 100.0%, respectively. The inter-day and intra-day variation coefficients were all less than 10% (n=5).CONCLUSION The method developed is applied successfully to study the metabolism of palonosetron in human liver microsomes.