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Simultaneous determination and determination of sildenafil and its active metabolite in human plasma using LC-MS/MS method.
Biomedical Chromatography ( IF 1.8 ) Pub Date : 2020-06-20 , DOI: 10.1002/bmc.4927
Lin-Ling Li 1 , Xiu-Jin Zhang 2 , Chao Zheng 2 , Ming-Xia Fan 2 , Hao-Yun Xu 2 , Jin-Ling Yi 3 , Yan Feng 3 , Xi Luo 2 , Wei-Hua Li 2 , Ze-Neng Cheng 1
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A sensitive and selective high‐performance liquid chromatography–tandam mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous quantification of sildenafil and its metabolite N‐desmethyl sildenafil in human plasma. Sildenafil‐d8 was used as an internal standard. The analytes were extracted by precipitation extraction and chromatographed on a C18 column using mobile phase A of water (containing 0.1% formic acid) and mobile phase B of acetonitrile (containing 0.1% formic acid) with gradient elution. Quantification was done using multiple reaction monitoring mode to monitor the precursor‐to‐product ion transitions of m/z 475.4 → m/z 283.3 for sildenafil, m/z 461.4 → m/z 283.2 for N‐desmethyl sildenafil and m/z 483.3 → m/z 108.1 for IS in positive ionization mode. The calibration curve was established over the range of 2.00–1,000 ng/ml and the correlation coefficient was >0.99. The intra‐day and inter‐day relative standard deviations were <6.5% for sildenafil and 6.3% for N‐desmethyl sildenafil respectively. Accuracy determinaed at four concentrations was 86.50–105.67% for sildenafil and 96.83–114.40% for N‐desmethyl sildenafil. This method was successfully applied to a pharmacokinetic description of sildenafil and the effect of food intake on the pharmacokinetics of sildenafil was also demonstrated in healthy Chinese volunteers.
更新日期:2020-06-20
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