BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF REMDESIVIR IN HUMAN PLASMA BY LC-MS/MS AS PER ICH M10 GUIDELINE

SUBHRANSHU PANDA; TUSHAR CHAVAN; RAVINDRA BHAVSAR

doi:10.22159/ijap.2025v17i6.55897

Authors

SUBHRANSHU PANDA School of Pharmaceutical Sciences, Jaipur National University, Jagatpura, Jaipur, Rajasthan-302017, India https://orcid.org/0000-0003-4782-8895
TUSHAR CHAVAN School of Pharmaceutical Sciences, Jaipur National University, Jagatpura, Jaipur, Rajasthan-302017, India https://orcid.org/0009-0008-7137-2879
RAVINDRA BHAVSAR Department of Clinical, Pharmadesk Solutions Pvt Ltd, Navi Mumbai, Maharashtra-400710, India

DOI:

https://doi.org/10.22159/ijap.2025v17i6.55897

Keywords:

Remdesivir, Bioanalytical, Method validation, Bioanalysis, Clinical study, ICHM10, LC-MS/MS

Abstract

Objective: Remdesivir, an adenosine analogue prodrug widely used during Coronavirus disease 2019 (COVID-19) pandemic. In this research, the objective was to develop a cost-effective and robust bioanalytical method capable of accurately quantifying remdesivir in human clinical studies as per ICH M10 guideline.

Methods: Development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with electro spray ionisation (ESI) source primarily involved extensive chromatographic optimization trials. Among various tested columns, Waters Symmetry C18 (5 µm, 3.9 mm×150 mm) demonstrated most consistent and well-defined chromatographic response when used with a highly organic acidified mobile phase, achieving a short run time of 3 min. For extraction method optimization, multiple approaches were evaluated, with protein precipitation emerging as preferred technique with environmentally friendly reagent use.

Results: Bioanalytical method validation was performed well as per ICH M10 (International Council for harmonisation of Technical Requirements for pharmaceuticals for Human Use) guidelines. The method demonstrated good linearity from 50-5000 ng/ml, with a correlation coefficient (R²) of 0.9978. The within-run and between-run precision for all quality control (QC) levels remained up to 6.65%, while accuracy ranged from 95.22-110.74%. Remdesivir was stable under freeze-thaw, auto sampler, bench top and long-term settings, with a percentage change of±15%. The paired t-test analysis revealed no significant difference between fresh and stability samples (p>0.1).

Conclusion: Validated calibration range of 50–5000 ng/ml is suitable for bioanalysis of remdesivir in human clinical studies involving intravenous infusion, considering reported Cmax of approximately 2229 ng/ml. This study highlights the integration of advanced LC-MS/MS techniques for quantification of remdesivir using remdesivir D5 as an internal standard in human plasma.

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