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.
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™.
State University of Campinas, Campinas, SP, Brazil.
Celso Francisco Pimentel
CF
Vespasiano
State University of Campinas, Campinas, SP, Brazil.
Tiago Luders
TL
Laurito
State University of Campinas, Campinas, SP, Brazil.
Renan Donomae
RD
Iwamoto
Galeno Research Unit, Campinas, SP, Brazil.|Faculty of Health Sciences of Barretos, Barretos, Brazil.
Ronilson Agnaldo
RA
Moreno
State University of Campinas, Campinas, SP, Brazil.|Galeno Research Unit, Campinas, SP, Brazil.|University Camilo Castelo Branco, São Paulo, SP, Brazil.
Gustavo D
GD
Mendes
State University of Campinas, Campinas, SP, Brazil.|Galeno Research Unit, Campinas, SP, Brazil.|University Camilo Castelo Branco, São Paulo, SP, Brazil.
Gilberto
G
De Nucci
D000328:Adult; D019540:Area Under Curve; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D018592:Cross-Over Studies; D004338:Drug Combinations; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D006852:Hydrochlorothiazide / Q000097:blood / Q000493:pharmacokinetics*; D008297:Male; D008875:Middle Aged; D000068577:Nebivolol / Q000097:blood / Q000493:pharmacokinetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards; D013810:Therapeutic Equivalency; D014147:Tramadol / Q000097:blood
D004338:Drug Combinations; D000068577:Nebivolol; D006852:Hydrochlorothiazide; D014147:Tramadol
LC-MS/MS; bioequivalence study; human plasma; hydrochlorothiazide; nebivolol