Objective:
The aim of the present work is to achieve a novel highly sensitive chromatographic
method for the simultaneous determination of hepatitis C agents, sofosbuvir and velpatasvir
from human plasma using ritonavir as an internal standard.
Methods:
Chromatographic separation was achieved using Hypersil C18 column (50mm x 4.6mm, 3μm)
with an isocratic elution mode using the mobile phase composition 10 mM ammonium formate buffer
(pH 5.0): acetonitrile (20:80 v/v) pumped at a flow rate of 0.5 ml/min. The detection was carried out by
tandem mass spectrometry using Multiple Reaction Monitoring (MRM) positive Electrospray Ionization
(ESI) with proton adducts at m/z 530.10 > 243.10, 883.40 > 114.0 and 721.25 > 197.0.
Results:
The method validated as per USFDA guidelines with respect to linearity, accuracy, and precision
was found to be acceptable over the concentration range of 0.2–2000 ng/ml and 5-2000 ng/ml for
sofosbuvir and velpatasvir respectively and the method was found to be highly sensitive and selective.
Conclusion:
The developed tandem mass spectrometric method is robust and can be applied for the
monitoring of plasma levels of the analyzed drug in preclinical and clinical pharmacokinetic studies.
Highly sensitive and selective analysis of urinary steroids by comprehensive two-dimensional gas chromatography combined with positive chemical ionization quadrupole mass spectrometry
