Liquid Chromatography-Tandem Mass Spectrometry Method for the Determination of Vardenafil and Its Application of Bioequivalence.

Kok Zheng Gan,Mohd Izwan Mohamad Yusof,Riyanto Teguh Widodo,Lay Kek Teh,Mohd Salleh Rofiee,Zamri Chik

doi:10.1155/2021/5590594

Abstract

A simple, rapid, and sensitive method of liquid chromatography-tandem mass spectrometry (LC/MS/MS) method was developed and validated for the determination of vardenafil in rabbit plasma. A simple protein precipitation method with ice-cold acetonitrile was used for plasma extraction. The mass transitions m/z 489⟶151 and m/z 390⟶169 were used to measure vardenafil and tadalafil (internal standard), respectively, with a total assay run time of 6 min. The limit of detection was 0.2 ng/mL. The assay was reproducible with intra-assay and interassay precision ranging 1.17%–9.17% and 1.31%–5.86%, respectively. There was also good intra-assay and interassay accuracy between 89.3%–105.3% and 94%–102% of the expected value, respectively. The linearity range was 0.5–60 ng/mL in rabbit plasma (r2 ≥ 0.99). The measured AUC from 0 to 24 h (AUC0 − 24t) for the test and reference formulations were 174.38 ± 95.91 and 176.45 ± 76.88, respectively. For the test, Cmax and Tmax were 75.36 ± 59.53 ng/mL and 1.42 ± 0.19 h, whereas, for the reference, these were 58.22 ± 36.11 ng/mL and 2.04 ± 0.33 h, respectively. The test formulation achieved a slightly lower AUC0 − 24t value (p > 0.05), higher Cmax values (p > 0.05), faster Tmax (p < 0.05), and almost equal bioavailability compared with the reference formulation.

Highlights

Vardenafil (2-[2-ethoxy-5-(4-ethylpiperazin-1-yl)sulfonylphenyl]-5-methyl-7-propyl-3H-imidazo [5, 1-f] [1, 2, 4] triazin-4-one; trihydrate; hydrochloride) is a highly potent and selective phosphodiesterase 5 (PDE5) inhibitor [1]. is is generally well-tolerated in men with erectile dysfunction (ED) with no reports of abnormal color vision at recommended doses (5–20 mg) [2]
No interfering peak was observed at the retention time of the analytes
Blank plasma spiked with low limit of quantification (LLOQ) and upper limit of quantification (ULOQ) concentrations of vardenafil and Internal Standard (IS) are illustrated in Figures 3(b) and 3(c), respectively

Summary

Introduction

Vardenafil (2-[2-ethoxy-5-(4-ethylpiperazin-1-yl)sulfonylphenyl]-5-methyl-7-propyl-3H-imidazo [5, 1-f] [1, 2, 4] triazin-4-one; trihydrate; hydrochloride) is a highly potent and selective phosphodiesterase 5 (PDE5) inhibitor [1]. is is generally well-tolerated in men with erectile dysfunction (ED) with no reports of abnormal color vision at recommended doses (5–20 mg) [2]. Vardenafil (2-[2-ethoxy-5-(4-ethylpiperazin-1-yl)sulfonylphenyl]-5-methyl-7-propyl-3H-imidazo [5, 1-f] [1, 2, 4] triazin-4-one; trihydrate; hydrochloride) is a highly potent and selective phosphodiesterase 5 (PDE5) inhibitor [1]. E mean half-life (t1/2) of vardenafil ranges from 3.94 to 4.79 h [2]. Vardenafil is highly bound to plasma protein (95%). It is extensively metabolized in the liver mainly by cytochrome P450 (CYP) 3A4 and a smaller portion by CYP3A5 and CYP2C isoforms [3]. Vardenafil film-coated tablets undergo extensive first-pass metabolism, and their absolute bioavailability is approximately 15% [4]

Results

Discussion

Conclusion