UPLC-Q-TOF-MS法快速检测补肾壮阳类中成药中11个PDE5型抑制剂类药物

目的：建立补肾壮阳类中药中11个磷酸二酯酶5型（PDE5型）抑制剂类药物的超高效液相色谱-四极杆飞行时间质谱（UPLC-Q-TOF-MS）快速检测方法。方法：样品经乙腈超声提取后，应用Agilent Extend-C₁₈（1.8 μm，2.1 mm×50 mm）色谱柱，以乙腈（A）-含0.1%醋酸的5 mmol·L^-1醋酸铵水溶液（B）为流动相，梯度洗脱（0~3.5 min，27%A；3.5~12 min，27%A→69%A；12~14 min，69%A；14~20 min，27%A），流速0.20 mL·min^-1，采用四极杆飞行时间质谱（Q-TOF-MS）检测器，电喷雾离子源（ESI），毛细管电压（capillary voltage）为4.0 kV，碎裂电压（Fragmentor）为180 V，正离子全扫描检测，扫描范围为m/z 100~1500。结果：那红地那非、红地那非、伐地那非、羟基豪莫西地那非、西地那非、豪莫西地那非、氨基他达拉非、他达拉非、硫代艾地那非、伪伐地那非、那莫西地那非11个化合物14 min内即可完成检测，在50~10000 μg·L^-1范围内线性相关系数均大于0.999；10倍定量限（LOQ）添加水平的加样回收率在93.8%~102.1%之间，RSD在2.6%~4.8%之间。结论：该方法操作简便、快速，经方法学验证可用于中成药中非法添加PDE5型抑制剂的快速检测。

Objective: To develop an ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometric (UPLC-Q-TOF-MS) method for the determination of 11 phosphodiesterase-5 inhibitors in Chinese medicine preparations for invigorants. Methods: Samples were extracted by ultrasonic method with acetonitrile,and then analyzed by UPLC-Q-TOF-MS.The Agilent Extend-C₁₈ column (1.8 μm,2.1 mm×50 mm) was adopted with the mobile phase consisting of acetonitrile(A)-5 mmol·L^-1 ammonium acetate water solution containing 0.1% acetic acid(B) with gradient elution (0-3.5 min,27%A;3.5-12 min,27%A→69%A;12-14 min,69%A;14-20 min,27%A) at a flow rate of 0.20 mL·min^-1 .The Q-TOF-MS was equipped with ESI source.The capillary voltage was 4 kV,and the fragmentor was 180 V,respectively.The detector was operated in positive ion mode by full scan range of m/z 100-1500. Results: Eleven PDE5 inhibitors were determined within 14 minutes,such as noracetildenafil,acetildenafil,vardenafil,hydroxyhomosildenafil,sildenafil,homosildenafil,aminotadalafil,tadalafil,thioaildenafil,pseudovardenafil,norneosildenafil.The correlation coefficients were all more than 0.999 with linear range of 50-10000 μg·L^-1.The average recovery of 11 PDE5 inhibitors at the spiked level of 10-fold LOQ was between 93.8% and 102.1% with the relative standard deviation between 2.6% and 4.8%. Conclusion: The method is simple and rapid,which can be applied to the detection of PDE5 inhibitors in Chinese medicine preparations according to methodology validation.