Author(s):
Srinivasa Reddy, Licto Thomas, Nirmala Nayak, Srinu Damatoti, Arindam Mukhopadhyay, Saral Thangam
Email(s):
arindam.mukhopadhyay@norwichclinical.com
DOI:
10.52711/2231-5691.2025.00004 
Address:
Srinivasa Reddy, Licto Thomas, Nirmala Nayak, Srinu Damatoti, Arindam Mukhopadhyay*, Saral Thangam
Norwich Clinical Services Pvt. Limited, 147/F, 8th Main, 3rd Block, Koramangala, Bangalore – 560 034, Karnataka, India.
*Corresponding Author
Published In:
Volume - 15,
Issue - 1,
Year - 2025
ABSTRACT:
A method for quantifying Baloxavir in human plasma using liquid chromatography-tandem mass spectrometry (LCMS/MS) with Baloxavir-d5 as an internal standard was detailed. Sample preparation involved liquid-liquid extraction, and chromatographic separation utilized an Acquity UPLC Peptide BEH C18 Column (300Å, 1.7µm, 2.1mm x 150mm). Positive mode monitoring of multiple reaction monitoring (MRM) transitions included m/z 484.1?247.0 for Baloxavir and m/z 489.1?252.1 for Baloxavir-d5. Sample concentrations were determined through linear regression analysis using Analyst software version 1.7.2, demonstrating an excellent linear response within the concentration range of 0.505 to 302.724 ng/ml for Baloxavir. Intra-day and inter-day precision were both within 3.95% for bulk spiked PA batches. The assay accuracy ranged from 97.08% to 105.51% for intra-batch and from 97.49% to 101.99% for inter-batch bulk spiked PA samples. Mean recoveries were 81.29% for Baloxavir and 92.76% for Baloxavir-d5. The limit of detection for Baloxavir was 0.127ng/ml. This methodology proved successful in a bioequivalence study.
Cite this article:
Srinivasa Reddy, Licto Thomas, Nirmala Nayak, Srinu Damatoti, Arindam Mukhopadhyay, Saral Thangam. Asian Journal of Pharmaceutical Research. 2025; 15(1):13-1. doi: 10.52711/2231-5691.2025.00004
Cite(Electronic):
Srinivasa Reddy, Licto Thomas, Nirmala Nayak, Srinu Damatoti, Arindam Mukhopadhyay, Saral Thangam. Asian Journal of Pharmaceutical Research. 2025; 15(1):13-1. doi: 10.52711/2231-5691.2025.00004 Available on: https://asianjpr.com/AbstractView.aspx?PID=2025-15-1-4
REFERENCES:
1. Carrat F, Vergu E, Ferguson NM, Lemaitre M, Cauchemez S, Leach S, et al. Time lines of infection and disease in human influenza: a review of volunteer challenge studies. Am J Epidemiol. 2008; 167(7): 775-85.
2. Wright PF, Neumann G, Kawaoka Y. Orthomyxoviruses. In: Knipe DM, Howley PM, eds. Fields Virology. 5th edn. LippincottWilliams and Wilkins, 2007; 1691–740.
3. Taubenberger JK, Morens DM. The pathology of influenza virus infections. Annu Rev Pathol. 2008; 3: 499-522.
4. Fukao K, Noshi T, Yamamoto A, Kitano M, Ando Y, Noda T, et al. Combination treatment with the cap-dependent endonuclease inhibitor baloxavir marboxil and a neuraminidase inhibitor in a mouse model of influenza A virus infection. J Antimicrob Chemother. 2019; 74(3): 654-62.
5. Lackenby A, Besselaar TG, Daniels RS, Fry A, Gregory V, Gubareva LV, et al. Global update on the susceptibility of human influenza viruses to neuraminidase inhibitors and status of novel antivirals, 2016-2017. Antiviral Res. 2018; 157: 38-46.
6. Bosaeed M, Kumar D. Seasonal influenza vaccine in immunocompromised persons. Hum Vaccin Immunother. 2018; 14(6): 1311-22.
7. Omoto S, Speranzini V, Hashimoto T, Noshi T, Yamaguchi H, Kawai M, et al. Characterization of influenza virus variants induced by treatment with the endonuclease inhibitor baloxavir marboxil. Scientific Reports. 2018; 8.
8. Jancy Rani E. H3N2 Influenza Virus: Is it Pandemic? A and V Pub International Journal of Nursing and Medical Research. 2023:56-9.
9. Kengar M, Jadhav A, Jadhav R, Jarag P, Shete S. Swine Flu and its Recent Medicines. Asian Journal of Pharmaceutical Analysis. 2019; 9: 30.
10. Hayden FG, Sugaya N, Hirotsu N, Lee N, de Jong MD, Hurt AC, et al. Baloxavir Marboxil for Uncomplicated Influenza in Adults and Adolescents. N Engl J Med. 2018; 379(10): 913-23.
11. Reddy S, Nayak N, Ahmed I, Thomas L, Mukhopadhyay A, Thangam S. Development and Validation of two LCMS/MS Methods for Simultaneous Estimation of Oseltamivir and its Metabolite in Human Plasma and Application in Bioequivalence Study. Asian Journal of Pharmaceutical Analysis. 2016; 6: 91.
12. Koshimichi H, Ishibashi T, Kawaguchi N, Sato C, Kawasaki A, Wajima T. Safety, Tolerability, and Pharmacokinetics of the Novel Anti-influenza Agent Baloxavir Marboxil in Healthy Adults: Phase I Study Findings. Clin Drug Investig. 2018; 38(12): 1189-96.
13. A New Stability Indicating RP-HPLC Method Development And Validation For Estimation Of Baloxavir Marboxil In Pharmaceutical Dosage Form. Journal of Pharmaceutical Negative Results. 2022: 970-83.
14. Gouda AS, Abdel-Megied AM, Rezk MR, Marzouk HM. LC-MS/MS-based metabolite quantitation of the antiviral prodrug baloxavir marboxil, a new therapy for acute uncomplicated influenza, in human plasma: Application to a human pharmacokinetic study. J Pharm Biomed Anal. 2023; 223: 115165.
15. Guidance for Industry, Bioanalytical Method Validation, U.S. Department of Health and Human Services Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM) May 2018, Available form: https://www.fda.gov/files/drugs/published/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf
16. Khan H, Ali J. UHPLC: Applications in Pharmaceutical Analysis. Asian Journal of Pharmaceutical Analysis. 2017; 7: 124.
17. Yousra Y, Fatima S, Rasheed N, Mohammad A. A Brief Review on Fundamentals of Analytical Chemistry. Asian Journal of Research in Pharmaceutical Science. 2017; 7: 13.
18. Shashikala P, Dodda S, Bakshi V. Development and Validation of Bioanalytical Method for the Estimation of Carisoprodol in Human Plasma using LC-MS/MS. Asian Journal of Pharmaceutical Analysis. 2015; 5: 181.
19. Khan H. Analytical Method Development in Pharmaceutical Research: Steps involved in HPLC Method Development. Asian Journal of Pharmaceutical Analysis. 2017; 7: 203.
20. Amruta S. Kadam, Nayana V. Pimpodkar, Puja S. Gaikwad, Sushila D. Chavan. Bioanalytical Method Validation. Asian Journal of Pharmaceutical Analysis. 2015; 5(4): 219-25.
21. Sahoo CK, Sudhakar M, Ramana DV, Satyanarayana K, Panda KC. Validation of Analytical Procedures- A Review. Asian Journal of Pharmaceutical Analysis. 2018; 8 (2): 109.
22. Saudagar R. B., Thete P. G. Bioanalytical Method Validation: A Concise Review. Asian Journal of Pharmaceutical Analysis, 2018; 8(2): 107-114