Validated isocratic stability-indicating RP-HPLC method for concurrent remdesivir estimation in drug products used in bioavailability studies | [Pakistan Journal of Pharmaceutical Sciences • 2025]

Author(s):

1. Muhammad Ibrahim: Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Government College University Faisalabad,,Pakistan

2. Muhammad Irfan: Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Government College University Faisalabad,,Pakistan

3. Syed Haroon Khalid: Department of Pharmaceutical Technology, Faculty of Pharmacy, Universiti Teknologi Mara (UiTM), Puncak Alam, Selangor, Malaysia; Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Government College University Faisalabad, Universiti Teknologi Mara (UiTM), Puncak Alam, Selangor, Pakistan

Abstract:

This study presents a stability-indicating, isocratic RP-HPLC method for the quantitative analysis of remdesivir (RMD) in pharmaceutical formulations and bioavailability studies. The method was developed to ensure robustness, precision and accuracy across diverse matrices, including freeze-dried powders, microspheres, novasomes and biological samples like blood. Chromatographic separation was achieved using a Zorbax Eclipse Plus C-18 column (4.6 × 250 mm, 5 µm) with a mobile phase of 1% trifluoroacetic acid (TFA) and acetonitrile (55:45) at a 1 mL/min flow rate. Detection was carried out at 254 nm using a variable wavelength detector (VWD). Validation followed current good manufacturing practices (cGMP), covering parameters such as system suitability, linearity, specificity, precision, accuracy, robustness and forced degradation. The method showed a retention time consistent with RMD standards, a limit of detection (LOD) of 0.73 µg/mL and a limit of quantification (LOQ) of 2.22 µg/mL. Excellent linearity (R² = 0.999) was observed in the 12-120 µg/mL range. Long-term and accelerated stability studies confirmed the method's capability to detect degradation products without interference. The validated method supports reliable routine quality control testing and pharmacokinetic studies, making it suitable for in-vivo bioavailability assessments of RMD formulations.

Page(s): 2120-2128

DOI: 10.36721/PJPS.2025.38.6.REG.14360.1

Published: Journal: Pakistan Journal of Pharmaceutical Sciences, Volume: 38, Issue: 6, Year: 2025

Keywords:

RPHPLC , Remdesivir , Stability Indicating Method

References:

[1] Abdul Samad H,Nadir Ali S,Qayoom A,Haroon U,Saad M,Mumtaz Alavi G .2023 .COVID-19 pandemic: perspectives on an unfolding crisis. Pak. J. Pharm. Sci., 36(1) : 159-170.

[2] Abo-Gharam AH,El-Kafrawy DS .2022 .Ecofriendly stability-indicating HPTLC microdetermination of the first FDA approved SARS-CoV-2 antiviral prodrug Remdesivir: Study of degradation kinetics and structural elucidation of the degradants using HPTLC-MS. Sustan. Chem, 29 : 100744.

[3] Anitha A,Rajamohan R,Murugan M,Seo JH M .2024 .Inclusion complexation of remdesivir with cyclodextrins: A comprehensive review on combating coronavirus resistance-Current state and future perspectives. Molecules, 29(19) : 4782.

[4] Arayne MS,Sultana N,Hussain F .2011 .Validated RP-HPLC method for determination of permethrin in bulk and topical preparations using UV-vis detector. J. Chromatogr. Sci., 49(4) : 287-291.

[5] Avataneo V,de Nicolo A,Cusato J,Antonucci M,Manca A,Palermiti A,D'Avolio A .2020 .Development and validation of a UHPLC-MS/MS method for quantification of the prodrug remdesivir and its metabolite GS-441524: A tool for clinical pharmacokinetics of SARS-CoV-2/COVID-19 and Ebola virus disease. J. Antimicrob. Chemother., 10(7) : 1772-1777.

[6] Bakheit AH,Darwish H,Darwish IA,Al-Ghusn AI .2023 .Remdesivir. Profiles of drug substances, excipients. , 48 : 71-108.

[7] Bhujbal S,Rupenthal ID,Agarwal P .2024 .Development and validation of stability-indicating HPLC method for assay of tonabersat in pharmaceutical formulations. Methods, 231 : 178-185.

[8] Cucinotta D and Vanelli M .2020 .WHO declares COVID-19 a pandemic. Acta bio medica: Atenei parmensis, 91(1) : 157.

[9] Gilead R .2020 .. Summary on Compassionate Use, : .

[10] Gupta PK,Pancholi SS,Das P .2024 .In-vitrodrug release testing method for nepafenac ophthalmic suspension. J.Pharm. Sci., 113(4) : 1061-1067.

[11] Hoseininezhad-Namin MS,Rahimpour E,Jouyban A .2024 .Favipiravir, remdesivir and lopinavir: Metabolites, degradation products and their analytical methods. Drug Metab. Rev., 56(2) : 127-144.

[12] Illendula S,Sindhuja PL,Suresh CV . .Rao K (2023b). A Novel Val.dated RP-HPLC method for the estimation of remdesivir in bulk and pharmaceutical dosage form. Int. J. Adv. Res. Med. Pharm. Sci. (IJARMPS), 8(1) : 58-67.

[13] Jubele A .2018 .Chromatographic determination of amines in food samples. M. Sc. , : .

[14] Mahase E and Kmietowicz Z .2020 .Doctors are told not to perform CPR on patients in cardiac arrest: BMJ 368-m1282. , 19 : .

[15] Mohyeldin SM,Daabees HG,Talaat W,Kamal MF .2024 .Efficient integration of analytical quality-bydesign and tri-hued HPLC-DAD approach for synchronized assessment of gemigliptin and rosuvastatin in their fixed-dose pills: Application to stability study. Sustain. Chem, 42 : 101777.

[16] Nyarko RO,Boateng E,Kahwa I,Boateng PO .2020 .A comparison analysis on remdesivir, favipiravir, hydroxychloroquine, chloroquine and azithromycin in the treatment of corona virus disease 2019 (COVID19)-A review. World J. Pharm. Pharm. Sci., 9(5) : 121-133.

[17] Palermiti A,De Nicolo A,Antonucci M,Soloperto S,Billi M,Manca A,Calcagno A .2025 .A novel HPLCMS/MS method for the intracellular quantification of the active triphosphate metabolite of remdesivir: GS443902. , 15(4) : 107.

[18] Patel V,Tiwari N,Patel K .2021 .Stability indicating RP-HPLC method development and validation for the estimation of Remdesivir in API form. World J. Pharm. Pharm. Sci., 10 : 1544-1551.

[19] Paul M,Astier A,Vieillard V .2018 .Extended stability of a biosimilar of trastuzumab (CT-P6) after reconstitution in vials, dilution in polyolefin bags and storage at various temperatures. GaBI J, 7(3) : 101-110.

[20] Poston JT,Patel BK,Davis AM .2020 .Management of critically ill adults with COVID-19. , 323(18) : 1839-1841.

[21] Qiu C,Zhang Y,Fan Y,Li S,Gao J,He X,Zhao X .2024 .Solid dispersions of genistein via solvent rotary evaporation for improving solubility, bioavailability and amelioration effect in HFD-induced obesity mice. Pharmaceutics, 16(3) : 306.

[22] Raasi KM .2021 .Analytical method development and validation of Remdesivir in bulk and pharmaceutical dosage forms using reverse-phase-high performance liquid chromatography. Int. J. Phram. Sci. Clin. Res., 1(2) : 282-329.

[23] Rodriguez-Morales AJ,Cardona-Ospina JA,GutiérrezOcampo E,Villamizar-Peña R,Holguin-Rivera Y,Escalera-Antezana JP,Henao-Martinez AF .2020 .Clinical, laboratory and imaging features of COVID19: A systematic review and meta-analysis. Travel Med, 34 : 101623.

[24] Tayeb HH,Felimban R,Almaghrabi S,Hasaballah N S .2021 .Nanoemulsions: Formulation, characterization, biological fate and potential role against COVID-19 and other viral outbreaks. Colloid. Interface. Sci. Commun, 45 : 100533.

[25] Wang M,Cao R,Zhang L,Yang X,Liu J,Xu M,Xiao G .2020 .Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019- nCoV) in-vitro. Cell Research, 30(3) : 269-271.

[26] Weiss P and Murdoch DR .2020 .Clinical course and mortality risk of severe COVID-19. , 395(10229) : 1014-1015.

[27] Weiss SR,Leibowitz JL SR .2011 .Coronavirus pathogenesis. Adv.Virus Res., 81 : 85-164.

[28] Wu C,Chen X,Cai Y,Zhou X,Xu S .2020 .Risk factors associated with acute respiratory distress syndrome and death in patients with coronavirus disease 2019 pneumonia in Wuhan, China. JAMA Internal Medicine, 180(7) : 934-943.

[29] Yuan M,Hu W,Feng Y,Tong Y,Wang X,Tan B,Liu J .2022 .Development and validation of an LC-MS/MS method for simultaneous determination of remdesivir and its hydrolyzed metabolite and nucleoside and its application in a pharmacokinetic study of normal and diabetic nephropathy mice. Biomed, 36(7) : e5380.

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