Department of Analytical Chemistry, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Vitoria-Gasteiz, Spain.
J Pharm Biomed Anal. 2012 Nov;70:529-33. doi: 10.1016/j.jpba.2012.04.030. Epub 2012 May 10.
This work proposes a liquid chromatography-electrospray ionization ion trap mass spectrometry (LC-ESI-ITMS) method, for the quantification of sildenafil (SDF), tadalafil (TDF) and vardenafil (VDF) and their metabolites N-desmethylSDF, O-desethylSDF and N-desethylVDF, preceded by a sample preparation step based on protein and phospholipid elimination. A C8 column (150 mm × 4.6 mm, 5 μm) with ammonium formate (20mM) and acetonitrile as the mobile phase components have been used. This method has been validated, obtaining limits of quantification ranged from 1 to 2.5 ng/mL and 2 to 5 ng/g in serum and brain tissue respectively, while limits of detection ranged from 0.3 to 0.9 ng/mL in serum and 0.6 to 1.9 ng/g in brain tissue. Assay recoveries for low level QC samples were higher than 83% and the matrix effect ranged between 91% and 108% in serum and between 98% and 107% in brain tissue. The method has been applied to the quantification of these compounds in the serum and brain tissue of rats treated intraperitoneally with 10 mg/kg of SDF, TDF or VDF.
本工作提出了一种液相色谱-电喷雾电离离子阱质谱（LC-ESI-ITMS）方法，用于定量分析西地那非（SDF）、他达拉非（TDF）和伐地那非（VDF）及其代谢物 N-去甲基 SDF、O-去乙基 SDF 和 N-去乙基 VDF，该方法包括基于蛋白质和磷脂去除的样品前处理步骤。采用 C8 柱（150mm×4.6mm，5μm），以甲酸铵（20mM）和乙腈为流动相成分。该方法经过验证，在血清和脑组织中的定量下限分别为 1 至 2.5ng/mL 和 2 至 5ng/g，检测下限分别为 0.3 至 0.9ng/mL 在血清和 0.6 至 1.9ng/g 在脑组织中。低浓度 QC 样品的测定回收率高于 83%，基质效应在血清中为 91%至 108%，在脑组织中为 98%至 107%。该方法已应用于经腹腔给予 10mg/kg SDF、TDF 或 VDF 的大鼠血清和脑组织中这些化合物的定量分析。
