HPLC and LCMS/MS
based chromatographic screening method impurities of Daprodustat with degradation
kinetics, characterization and in silico toxicity
Bhagya Kumar Tatavarti, Rasheed Babu Shaik, Uppu Naga Babu, Shaheda Niloufer and
Govindarao Yedlapalli
Res. J. Chem. Environ.; Vol. 29(9); 30-40;
doi: https://doi.org/10.25303/299rjce030040; (2025)
Abstract
The present study reports a sensitive and stability-indicating HPLC method with
LC-MS/MS confirmation for the complete impurity profiling, degradation kinetics
and in silico toxicity assessment of daprodustat, a new oral hypoxia-inducible factor
prolyl hydroxylase (HIF-PH) inhibitor. The earlier research describes various methods
for the quantification of daprodustat in drug products and biological matrices suggest
a gap for its impurity profiling, degradation kinetics and toxicity evaluation under
one analytical approach. This study fills this gap by critically elaborating a reverse-phase
chromatographic technique on a LiChrospher RP-18 column by isocratic elution with
acetonitrile and aqueous formic acid (65:35 v/v), with baseline separation of daprodustat
from its known impurities and degradation products (DPs) with resolution values
of more than 2.0.
The forced degradation tests were performed according to ICH-recommended stress
conditions under which significant degradation was seen upon acidic, basic and thermal
stresses. The acidic stress, three different DPs were observed, one of which was
determined to be a primary amide derivative of daprodustat by LC-MS/MS analysis
(m/z 336). In the acidic degradation kinetics conformed to pseudo-first-order kinetics.
The method was validated according to ICH Q2(R1) guidelines for specificity, linearity,
sensitivity, precision, accuracy, robustness and system suitability. The technique
was highly sensitive with LODs of 0.033 μg/mL for daprodustat and 0.075 μg/mL for
impurities. The recovery was between 98% and 102%, with intra- and inter-day precision
(% RSD) being less than 2%. The technique was very specific and did not have any
interference from excipients or formulation matrices. Overall, this study provides
a comprehensive analytical framework for the quality control, safety assessment
and regulatory compliance of daprodustat that makes it highly applicable for routine
pharmaceutical analysis.
