Quantification of sofosbuvir and ledipasvir in human plasma by UPLC-MS/MS method: Application to fasting and fed bioequivalence studies.
A rapid and sensitive LC-MS/MS method was developed, optimized and validated for quantification of sofosbuvir (SF) and ledipasvir (LD) in human plasma using eplerenone as an internal standard (IS). Analytes and IS were extracted from plasma by simple liquid-liquid extraction technique using methyl tertiary butyl ether. The prepared samples were chromatographed on Acquity UPLC BEH C18 column. Separation was done using a mobile phase formed of 0.1% formic acid and acetonitrile (50:50, v/v) in an isocratic mode at a flow rate of 0.4ml/min. The Xevo TQD LC-MS/MS was operated under the multiple-reaction monitoring mode using electrospray ionization. A full validation of the method was performed according to the FDA guidelines. Linearity was found to be in the range of 0.25-3500ng/ml for SF and 5-2000ng/ml for LD. The intra-day and inter-day precision and accuracy results were within the acceptable limits. A short run time of 2min allows analysis of more than 400 plasma samples per day. The developed method was successfully applied to both fasting and fed bioequivalence studies in healthy human volunteers.
Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562, Cairo, Egypt. Electronic address: mamdouh.rezk@pharma.cu.edu.eg.
Mamdouh R
MR
Rezk
Clinical Pharmacy Department, Faculty of Pharmaceutical Sciences & Pharmaceutical Industries, Future University in Egypt, Egypt.
Ehab R
ER
Bendas
Pharmaceutics and Industrial Pharmacy Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini Street, 11562, Cairo, Egypt.
Emad B
EB
Basalious
Advanced Research Center (ARC), Nasr City, Cairo, Egypt.
Iman A
IA
Karim
D000328:Adult; D000998:Antiviral Agents / Q000097:blood*; D001562:Benzimidazoles / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005215:Fasting; D005260:Female; D005449:Fluorenes / Q000097:blood*; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008875:Middle Aged; D000069474:Sofosbuvir / Q000097:blood*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013810:Therapeutic Equivalency; D055815:Young Adult
D000998:Antiviral Agents; D001562:Benzimidazoles; D005449:Fluorenes; C586541:ledipasvir; D000069474:Sofosbuvir
Bioequivalence; Ledipasvir; Plasma; Sofosbuvir; UPLC–MS/MS; Validation