Selective and rapid determination of tadalafil and finasteride using solid phase extraction by high performance liquid chromatography and tandem mass spectrometry

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 152; pp. 215 - 223
• Main Authors: Pappula, Nagaraju, Kodali, Balaji, Datla, Peda Varma
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 15.04.2018
• Subjects: Finasteride; Human plasma; Liquid chromatography–tandem mass spectrometry; Solid phase extraction; Tadalafil; Validation; Validation; Tadalafil; Solid phase extraction; Finasteride; Human plasma; Liquid chromatography–tandem mass spectrometry
• ISSN: 0731-7085; 1873-264X; 1873-264X
• DOI: 10.1016/j.jpba.2018.01.020

•A sensitive, selective and rapid method was developed for the simultaneous estimation of tadalafil and finasteride in human plasma.•Small plasma volume consumption of 250 μl, for sample preparation.•Method was fully validated as per USFDA, ANVISA, EMA guidelines.•Application to real subject samples for analysis of tadalafil and finasteride in a clinical study. Highly selective and fast liquid chromatography-tandem mass spectrometric (LC–MS/MS) method was developed and validated for simultaneous determination of tadalafil (TDL) and finasteride (FNS) in human plasma. The method was successfully applied for analysis of TDL and FNS samples in clinical study. The method was validated as per USFDA (United States Food and Drug Administration), EMA (European Medicines Agency), and ANVISA (Agência Nacional de Vigilância Sanitária-Brazil) bio analytical method validation guidelines. Glyburide (GLB) was used as common internal standard (ISTD) for both analytes. The selected multiple reaction monitoring (MRM) transitions for mass spectrometric analysis were m/z 390.2/268.2, m/z 373.3/305.4 and m/z 494.2/369.1 for TDL, FNS and ISTD respectively. The extraction of analytes and ISTD was accomplished by a simple solid phase extraction (SPE) procedure. Rapid analysis time was achieved on Zorbax Eclipse C18 column (50 × 4.6 mm, 5 μm). The calibration ranges for TDL and FNS were 5–800 ng/ml and 0.2–30 ng/ml respectively. The results of precision and accuracy, linearity, recovery and matrix effect of the method are acceptable. The accuracy was in the range of 92.9%–106.4% and method precision was also good; %CV was less than 8.1%.