超高效液相色谱-串联质谱法测定保健食品中10种磷酸二酯酶-5抑制剂

Simultaneous Determination of Ten Phosphodiesterase-5 Inhibitors in Health Foods by Ultra Performance Liquid Chromatography with Tandem Mass Spectrometry

建立了超高效液相色谱-串联质谱法（UPLC-MS/MS）同时测定保健食品中10种磷酸二酯酶-5（PDE-5）抑制剂。样品采用Waters Acquity UPLC BEH C18（100mm×2.1mm×1.7μm）色谱柱分离,以甲醇-0.1%甲酸水溶液为流动相进行梯度洗脱,甲醇为提取溶剂超声萃取,经HLB固相萃取柱净化浓缩;在电喷雾正离子模式下,以多反应监测（MRM）模式进行检测。结果表明,10种磷酸二酯酶-5抑制剂在1.0-100.0μg/L范围内线性关系良好,线性相关系数（R2）均大于0.993,方法定量限为1.0μg/kg;空白样品在100.0、200.0和1 000.0μg/kg添加水平下的回收率为85.6%-94.3%。该方法简便、快速、检出限低,适用于保健食品中10种磷酸二酯酶-5抑制剂的同时检测。

A method was developed for simultaneous determination of ten phosphodiesterase-5(PDE-5)inhibitors in health foods by ultra performance liquid chromatographytandem mass spectrometry(UPLC-MS/MS),such as vardenafil,acetildenafil,sildenafil,homosildenafil,udenafil,tadalafil,thioaildenafil,gendenafil,pseudovarnafil,nor-neosildenafil.The separation was performed using water containing 0.1% formic acid and methanol as mobile phases with gradient elution at a flow rate of 0.2 mL/min.Qualitative and quantitative analysis were achieved after the chromatographic separation on a Waters Acquity UPLC BEH C18column(100mm×2.1mm×1.7μm).The electrospray ionization(ESI)source in positive ion mode was used for analysis of ten phosphodiesterase-5inhibitors in the multiple reaction monitoring(MRM)mode.The nuclear ratio of parent ion and daughter ion,cone voltage,collision voltage were investigated.The sample was ultrasonic-assisted treatment using the methanol as extraction solvent,and then purified by Waters Oasis HLB solid-phase extraction(SPE)column.The results indicate that the linear range of the method is from 1.0μg/L to 100.0μg/L with R2>0.993,which means the method has a good linearity.The limits of detection(LOD)and limits of quantification(LOQ)of ten phosphodiesterase-5inhibitors are 0.3-1.0μg/kg and 1.0-2.5μg/kg,respectively.The average recoveries of ten phosphodiesterase-5inhibitors at three spiked levels of 100.0,200.0and 1 000.0μg/kg are 85.6%-94.3%.The method was applied to analysis of the investigated compounds in twenty health foods.The results indicate that sildenafil are found in eighteen samples with the range of 1.3-210.8 mg/g,vardenafil and tadalafil are found in a tablet and a capsule,respectively.This method is accurate,simple,rapid and feasible for simultaneous determination of phosphodiesterase-5inhibitors in health foods.