Simultaneous quantification of Daclatasvir and Sofosbuvir in human plasma and pharmacokinetic study by LCMS/MS

Jaiswal, Amarnath

Abstract

In the treatment of hepatitis C, direct-acting antivirals (DAA) are highly efficient and well tolerated with a series of DAA combinations available for treatment. A sensitive high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for the simultaneous quantification of Sofosbuvir (SOF) and Daclatasvir (DAC) in human plasma. Sofosbuvir D6 (SOF D6) and Daclatasvir 13C22H6 (DAC 13C22H6) are used as internal standard (IS). Quantification for both the analytes has been attained with MS-MS detection in positive ion mode using an Acquity UPLC system (Waters) equipped with Waters Xevo TQ MS system with a Gemini NX 5µ C18 (50 × 2.0mm) (Phenomenex) column, and a gradient mobile phase consisting 5 mM Ammonium Formate buffer: Acetonitrile at a flow rate of 0.300 mL/min is used as mobile phase to separate the analytes and detection is performed by electrospray ionization technique using the mass spectrometer. Full validation is performed for bio-analytical methods with respect to linearity, precision, accuracy, selectivity, carry-over, stability and dilution integrity. Linearity is obtained over a concentration range of 10.002 -3000.488 and 10.004 -3001.218 ng mL-1 for SOF and DAC respectively by applying weighted least-squares linear regression method (1/x2). The developed method was applied successfully in bioequivalence and/or clinical studies in 48 male subjects for the simultaneous quantification of SOF and DAC.




Keyword(s)

Bioequivalence; Daclatasvir; Good clinical practice; High performance liquid chromatography; Liquid Chromatography; Lower limit of quantification; Mass Spectrometry; Sofosbuvir


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