title: Development and validation of UFLC-MS/MS method for determination of bosentan in rat plasma.
abstract: A rapid, simple and sensitive UFLC-MS/MS method was developed and validated for the determination of bosentan in rat plasma using etodolac as an internal standard (IS) after liquid-liquid extraction with diethyl ether-chloroform (4:1, v/v). Bosentan and IS were detected using electrospray ionization in positive ion multiple reaction monitoring mode by monitoring the transitions m/z 551.90→201.90 and 288.20→172.00, respectively. Chromatographic separation was performed on the inertsil ODS-4 column with a gradient mobile phase, which consisted of 0.1% acetic acid with 5mM ammonium acetate in water for solvent A and 5mM ammonium acetate in acetonitrile-methanol (50:50, v/v) for solvent B at a flow rate of 0.3mL/min. The method was sensitive with 0.5ng/mL as the lower limit of quantitation (LLOQ) and the standard calibration curve for bosentan was linear (r>0.997) over the studied concentration range (0.5-2000ng/mL). The proposed method was fully validated by determining specificity, linearity, LLOQ, precision and accuracy, recovery, matrix effect and stability. The validated method was successfully applied to plasma samples obtained from rats.
authors.affiliation: Ataturk University, Department of Analytical Chemistry, Faculty of Pharmacy, Erzurum 25240, Turkey. Electronic address: alptugatila@yahoo.com.
authors.forename: Alptug
authors.initials: A
authors.lastname: Atila
authors.affiliation: Turkish Public Health Association, Directorate of Erzurum, Erzurum 25100, Turkey.
authors.forename: Murat
authors.initials: M
authors.lastname: Ozturk
authors.affiliation: Ataturk University, Department of Analytical Chemistry, Faculty of Pharmacy, Erzurum 25240, Turkey.
authors.forename: Yucel
authors.initials: Y
authors.lastname: Kadioglu
authors.affiliation: Ataturk University, Department of Pharmacology, Faculty of Medicine, Erzurum 25240, Turkey.
authors.forename: Zekai
authors.initials: Z
authors.lastname: Halici
authors.affiliation: Ataturk University, Department of Analytical Chemistry, Faculty of Pharmacy, Erzurum 25240, Turkey.
authors.forename: Didar
authors.initials: D
authors.lastname: Turkan
authors.affiliation: Ataturk University, Department of Pharmacology, Faculty of Medicine, Erzurum 25240, Turkey.
authors.forename: Muhammed
authors.initials: M
authors.lastname: Yayla
authors.affiliation: Ataturk University, Department of Biochemistry, Faculty of Pharmacy, Erzurum 25240, Turkey.
authors.forename: Harun
authors.initials: H
authors.lastname: Un
mesh_terms: D000818:Animals; D000077300:Bosentan; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D065128:Endothelin Receptor Antagonists / Q000097:blood*; D059625:Liquid-Liquid Extraction / Q000379:methods*; D008297:Male; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013449:Sulfonamides / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D065128:Endothelin Receptor Antagonists; D013449:Sulfonamides; D000077300:Bosentan
keywords: Bosentan; Liquid-liquid extraction; Rat plasma; UFLC–MS/MS; Validation