title: Development and validation of sensitive and rapid UPLC-MS/MS method for quantitative determination of daclatasvir in human plasma: Application to a bioequivalence study.
abstract: A rapid and sensitive UPLC-MS/MS method was developed and validated for determination of daclatasvir (DAC) in human plasma using sofosbuvir (SOF) as an internal standard (IS). The Xevo TQD LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. Precipitation with acetonitrile was used in sample preparation. The prepared samples were chromatographed on Acquity UPLC HSS C18 (50×2.1mm, 1.8μm) column by pumping 10mM ammonium formate (pH 3.5) and acetonitrile in an isocratic mode at a flow rate of 0.30ml/min. Method validation was performed as per the FDA guidelines and the standard curves were found to be linear in the range of 5-4000ng/ml for DAC. The intra-day and inter-day precision and accuracy results were within the acceptable limits. A very short run time of 1.2min made it possible to analyze more than 500 human plasma samples per day. The wider range of quantification of DAC allowed the applicability of the developed method for its determination in a bioequivalence study in human volunteers.
authors.affiliation: Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562, Cairo, Egypt. Electronic address: mamdouh.rezk@pharma.cu.edu.eg.
authors.forename: Mamdouh R
authors.initials: MR
authors.lastname: Rezk
authors.affiliation: Clinical Pharmacy Department, Faculty of Pharmaceutical Sciences & Pharmaceutical Industries, Future University in Egypt, Egypt.
authors.forename: Ehab R
authors.initials: ER
authors.lastname: Bendas
authors.affiliation: Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562, Cairo, Egypt.
authors.forename: Emad B
authors.initials: EB
authors.lastname: Basalious
authors.affiliation: Advanced Research Center (ARC), Nasr City, Cairo, Egypt.
authors.forename: Iman A
authors.initials: IA
authors.lastname: Karim
mesh_terms: D002219:Carbamates; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004355:Drug Stability; D006801:Humans; D007093:Imidazoles / Q000097:blood* / Q000737:chemistry*; D057230:Limit of Detection; D010949:Plasma / Q000737:chemistry*; D011759:Pyrrolidines; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D013810:Therapeutic Equivalency; D014633:Valine / Q000031:analogs & derivatives
chemical_list: D002219:Carbamates; D007093:Imidazoles; D011759:Pyrrolidines; D014633:Valine; C549273:daclatasvir
keywords: Bioequivalence; Daclatasvir; Pharmacokinetics; Plasma; UPLC–MS/MS; Validation