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Simultaneous determination and determination of sildenafil and its active metabolite in human plasma using LC–MS/MS method
Author(s) -
Li Linling,
Zhang Xiujin,
Zheng Chao,
Fan Mingxia,
Xu Haoyun,
Yi Jinling,
Feng Yan,
Luo Xi,
Li Weihua,
Cheng Zeneng
Publication year - 2020
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4927
Subject(s) - chemistry , sildenafil , chromatography , formic acid , desmethyl , pharmacokinetics , protein precipitation , metabolite , selected reaction monitoring , mass spectrometry , analyte , solid phase extraction , liquid chromatography–mass spectrometry , tandem mass spectrometry , pharmacology , biochemistry , medicine
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 C 18 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.