Bioequivalence study between a fixed‐dose single‐pill formulation of nebivolol plus hydrochlorothiazide and separate formulations in healthy subjects using high‐performance liquid chromatography coupled to tandem mass spectrometry

• Published in: Biomedical chromatography Vol. 31; no. 5; pp. np - n/a
• Main Authors: Vespasiano, Celso Francisco Pimentel, Laurito, Tiago Luders, Iwamoto, Renan Donomae, Moreno, Ronilson Agnaldo, Mendes, Gustavo D., De Nucci, Gilberto
• Format: Journal Article
• Language: English
• Published: England 01.05.2017
• Subjects: Adult; Area Under Curve; bioequivalence study; Calibration; Chromatography, High Pressure Liquid - methods; Chromatography, High Pressure Liquid - standards; Cross-Over Studies; Drug Combinations; Female; Healthy Volunteers; human plasma; Humans; hydrochlorothiazide; Hydrochlorothiazide - blood; Hydrochlorothiazide - pharmacokinetics; LC–MS/MS; Male; Middle Aged; nebivolol; Nebivolol - blood; Nebivolol - pharmacokinetics; Reproducibility of Results; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry - methods; Tandem Mass Spectrometry - standards; Therapeutic Equivalency; Tramadol - blood; LC-MS/MS; bioequivalence study; nebivolol; human plasma; hydrochlorothiazide
• ISSN: 0269-3879; 1099-0801
• DOI: 10.1002/bmc.3884

Systemic arterial hypertension is a major risk factor for cerebrovascular disease. Therefore, adequate control of blood pressure is of enormous importance. One of the many fixed‐dose single‐pill antihypertensive formulations available on the market is the combination of nebivolol and hydrochlorothiazide. The objective of this study was to develop two distinct high‐performance liquid chromatography coupled to tandem mass spectrometry methods to simultaneously quantify nebivolol and hydrochlorothiazide in human plasma. The methods were employed in a bioequivalence study, the first assay involving a nebivolol fixed‐dose single‐pill formulation based on healthy Brazilian volunteers. Nebilet HCT™ (nebivolol 5 mg + hydrochlorothiazide 12.5 mg tablet, manufactured by Menarini) was the test formulation. The reference formulations were Nebilet™ (nebivolol 5 mg tablet, manufactured by Menarini) and Clorana™ (hydrochlorothiazide 25 mg tablet, manufactured by Sanofi). For both analytes, liquid–liquid extraction was employed for sample preparation and the chromatographic run time was 3.5 min. The limits of quantification validated were 0.02 ng/mL for nebivolol and 1 ng/mL for hydrochlorothiazide. Since the 90% CI for Cmax, AUC(0–last) and AUC(0–inf) individual test/reference ratios were within the 80–125% interval indicative of bioequivalence, it was concluded that Nebilet HCT™ is bioequivalent to Nebilet™ and Clorana™.