Background:
Simeprevir is one of the recently discovered drugs for treating hepatitis C
which is one of the major diseases across the globe.
Objective:
The present study involves the development of a new and unique High-Performance Liquid
Chromatography (HPLC) method using fluorescence detection for the determination of simeprevir
(SIM) in human plasma.
Methods:
Two methods of extractions were tested, protein precipitation using acetonitrile and liquidliquid
extraction. A 25 mM dipotassium hydrogen orthophosphate (pH 7.0)/ACN (50/50; v/v), was
used as mobile phase and C18 reversed phase column as the stationary phase. The chromatographic
conditions were optimized and the concentration of simeprevir was determined by using the fluorescence
detector. Cyclobenzaprine was used as an internal standard.
Results:
Recovery of the assay method based on protein precipitation was up to 100%. Intra-day and
inter-day accuracies range from 92.30 to 107.80%, with Relative Standard Deviation (RSD) range
1.65-8.02%. The present method was successfully applied to a pharmacokinetic study where SIM
was administered as a single dose of 150 mg SIM/capsule (Olysio®) to healthy individuals.
Conclusion:
This method exhibits high sensitivity with a low limit of quantification 10 ng mL-1,
good selectivity using fluorescence detection, wide linear application range 10-3000 ng mL-1, good
recovery and highly precise and validation results. The developed method can be applied in routine
analysis for real samples.
