Quick and Sensitive UPLC-ESI-MS/MS Method for Simultaneous Estimation of Sofosbuvir and Its Metabolite in Human...

Area of Science:

• Analytical Chemistry
• Pharmacokinetics
• Biotechnology

Background:

• Accurate quantification of antiviral drugs and their metabolites is crucial for therapeutic drug monitoring.
• Sofosbuvir is a key component in Hepatitis C treatment, necessitating reliable analytical methods for its assessment.
• Existing methods may lack the sensitivity or throughput required for comprehensive pharmacokinetic studies.

Purpose of the Study:

• To develop and validate a sensitive, rapid, and robust RP-UPLC-MS/MS method for simultaneous determination of sofosbuvir and its major metabolite GS331007 in human plasma.
• To establish a method suitable for high-throughput bioanalysis in pharmacokinetic and bioequivalence studies.
• To ensure the method meets stringent regulatory validation requirements.

Main Methods:

• A reversed-phase ultra-performance liquid chromatography tandem mass spectrometry (RP-UPLC-MS/MS) technique was employed.
• Separation was performed on a C18 column with a mobile phase consisting of acetonitrile, ammonium formate, and formic acid.
• Sample preparation involved dichloromethane extraction, and analysis was conducted using positive electrospray ionization in multiple-reaction monitoring mode.

Main Results:

• The method demonstrated linearity over wide concentration ranges: 1-1000 ng/mL for sofosbuvir and 10-1500 ng/mL for GS331007.
• All validation parameters, including accuracy, precision, and selectivity, met FDA guidelines.
• Negligible matrix effects were observed, indicating minimal ion suppression or enhancement from plasma components.

Conclusions:

• The developed RP-UPLC-MS/MS method is simple, fast, and highly sensitive for quantifying sofosbuvir and GS331007 in human plasma.
• The method's robustness and validation compliance make it ideal for large-scale pharmacokinetic and bioequivalence studies.
• This assay provides a valuable tool for advancing research and clinical applications related to sofosbuvir therapy.