A GREENNESS EVALUATED BIOANALYTICAL METHOD FOR QUANTIFICATION OF SPARSENTAN IN RAT PLASMA AND ITS APPLICATION TO PHARMACOKINETIC STUDIES BY LC-MS

KRANTHI YALLA; A. KRISHNAMANJARI PAWAR; KIRAN MANDA

doi:10.22159/ijap.2025v17i5.54341

Authors

KRANTHI YALLA AU College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, Andhra Pradesh, India. Shri Vishnu College of Pharmacy, Bhimavaram, West Godavari, Andhra Pradesh, India https://orcid.org/0000-0003-4193-8129
A. KRISHNAMANJARI PAWAR AU College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, Andhra Pradesh, India https://orcid.org/0000-0002-2703-4505
KIRAN MANDA Shri Vishnu College of Pharmacy, Bhimavaram, West Godavari, Andhra Pradesh, India https://orcid.org/0000-0002-8289-2879

DOI:

https://doi.org/10.22159/ijap.2025v17i5.54341

Keywords:

Bench top stability, Macitentan, LC-MS, Cmax, Tmax, etc

Abstract

Objective: A simple, accurate, reproducible, greenness-evaluated method for bioanalytical estimation of sparsentan in rat plasma and its application to pharmacokinetic studies by liquid chromatography mass spectrometry (LC-MS) using macitentan as internal standard.

Methods: The current work sought to develop and estimate sparsentan by Chromatograph, Waters 2695 high performance liquid chromatography (HPLC) with auto sampler, column oven, and degasser, and SCIEX QTRAP 5500 mass spectrometer, and with class Empower-2 software. The Mobile Phase used was acetonitrile with trifluoroacetic acid buffer (50:50) with a run time of 5 min and a flow rate of 1 ml/min.

Results: Retention times of 1.933 for sparsentan and 3.270 for macitentan were found. This method was validated over a linear range of 6.0-240.0 ng/ml with a correlation coefficient of 0.9989. The intra-batch and inter-batch precision percentage coefficient of variation (%CV) across five levels of Quality parameters is less than 15.00. Bench top stability, short-term, and long-term stability studies were performed, and % CVs were found below 15%. Pharmacokinetic studies were performed by administering to rats and reported the maximum concentration (Cmax) and time to maximum concentration (Tmax). Green analysis was also performed.

Conclusion: The optimized sample preparation and chromatographic conditions ensured high sensitivity and minimal matrix interference, allowing reliable quantification of sparsentan even at low concentrations. Future research can focus on method modifications to enable simultaneous quantification of sparsentan and its metabolites.

References

1. Donadio JV, Grande JP. IgA nephropathy. N Engl J Med. 2002 Sep 5;347(10):738-48. doi: 10.1056/NEJMra020109, PMID 12213946.

2. Fabiano RC, Pinheiro SV, Simoes E Silva AC. Immunoglobulin a nephropathy: a pathophysiology view. Inflamm Res. 2016 Oct;65(10):757-70. doi: 10.1007/s00011-016-0962-x, PMID 27351940.

3. Lai KN, Tang SC, Schena FP, Novak J, Tomino Y, Fogo AB. IgA nephropathy. Nat Rev Dis Primers. 2016 Jan 14;2(1):16001. doi: 10.1038/nrdp.2016.1, PMID 27189177.

4. Floege J, Rauen T, Tang SC. Current treatment of IgA nephropathy. Semin Immunopathol. 2021 Oct;43(5):717-28. doi: 10.1007/s00281-021-00888-3, PMID 34495361.

5. Komers R, Plotkin H. Dual inhibition of rennin angiotensin aldosterone system and endothelin-1 in treatment of chronic kidney disease. Am J Physiol Regul Integr Comp Physiol. 2016 May 15;310(10):R877-84. doi: 10.1152/ajpregu.00425.2015, PMID 27009050.

6. Travere Therapeutics. Travere therapeutics announces FDA accelerated approval of FILSPARI™ (sparsentan), the first and only non-immunosuppressive therapy for the reduction of proteinuria in IgA nephropathy; 2023 Feb 17. Available from: https://www.travere.san.com/news/filsprai-fda-approvaldiego:traveretherapeutics. [Last accessed on 27 Feb 2024].

7. Travere Therapeutics Inc. FILSPARI™ (sparsentan) tablets for oral use: US prescribing information. San Diego: Travere Therapeutics; 2023.

8. Vifor CS, Travere Therapeutics. CSL vifor and travere therapeutics announce EMA has accepted for review the conditional marketing authorization application for sparsentan for the treatment of IgA Nephropathy; 2022 Aug 22. Available from: https://www.com/media-releases.viforpharma.

9. Travere Therapeutics. Travere therapeutics announces orphan drug designation for sparsentan for the treatment of IgA nephropathy. Available from: https://www.com/news/orphan-drug-sparsentan.travere. [Last accessed on 27 Feb 2024].

10. Drug Bank. Sparsentan-DB12548. Drug Bank. Available from: https://go.com/drugs/db12548.

11. Krishna PS, Mukkanti Eswarudu M, Srinivasa Babu P, Likhitha T, Venkatesh N, Poojitha C. A review on analytical profile for newly FDA-approved drugs in 2023. JMPAS. 2023;12(4):5955-61. doi: 10.55522/jmpas.V12I4.5491.

12. Siva Krishna P, Eswarudu MM, Srinivasa Babu P, Likhitha T, Venkatesh N, Poojitha C. Validated stability indicating RP-HPLC method for quantification of sparsentan in pharmaceutical dosage form: a 2023 USFDA approved drug. RJC. 2024 Apr-Jun;17(2):631-41. doi: 10.31788/RJC.2024.1728805.

13. Gali H. A new validated method for the estimation of sparsentan in bulk and pharmaceutical dosage form by RP-HPLC. World J Pharm Sci. 2024 Feb;12(2):47-58. doi: 10.54037/WJPS.2022.100905.

14. Wada R, Kleijn HJ, Zhang L, Chen SC. Population pharmacokinetic analysis of sparsentan in healthy volunteers and patients with focal segmental glomerulosclerosis. CPT Pharmacometrics Syst Pharmacol. 2023 Oct;12(8):1080-92. doi: 10.1002/psp4.12996, PMID 37221817.

15. Ramadevi P, Rambabu K. Bioanalytical method development and validation for ezetimibe and pitavastatin and its applications to pharmacokinetic studies in rat plasma by using LC-MS/MS. Int J Res Pharm Sci. 2020;11(4):7854-62. doi: 10.26452/ijrps.v11i4.4670.

16. Eluru A, Surendra Babu K. Bioanalytical method development and validation for aplidine in rat plasma and their pharmacokinetic studies by LC-MS. World J Pharm Pharm Sci. 2019;8:1201-9.

17. Ramchandran D, Kethipalli A, Krishnamurthy M. Bioanalytical method development and validation of daunorubicin and cytarabine in rat plasma by LC-MS/MS and its application in pharmacokinetic studies. J Pharm Sci Res. 2020;12(3):381-6.

18. US Food and Drug Administration. M10 bioanalytical method validation and study sample analysis. Silver Spring, MD: FDA; 2022 Nov.

19. European Medicines Agency. Guideline on bioanalytical method validation. Clinical orthopaedics and related research. Vol. 2. London, UK: EMA; 2011 Jul 21.

20. Gope ER, Pottendla S, Yaparthi S. Bioanalytical approach to ensitrelvir estimation using liquid chromatography tandem mass spectrometry and its application to pharmaceutical research. Asian J Pharm Clin Res. 2025 Mar;18(3):25-9. doi: 10.22159/ajpcr.2025v18i3.53760.

21. Hemanth Kumar AK, Sudha V, Vijayakumar A, Padmapriyadarsini C. Simultaneous method for the estimation of bedaquiline and delamanid in human plasma using high-performance liquid chromatography. Int J Pharm Pharm Sci. 2021;13(6):36-40. doi: 10.22159/ijpps.2021v13i6.40853.

22. Pena Pereira F, Wojnowski W, Tobiszewski M, Agree. Analytical greenness metric approach and software. Anal Chem. 2020 Aug 18;92(15):10076-82. doi: 10.1021/acs.analchem.0c01887.

23. Sajid M, Plotka Wasylka J. Green analytical chemistry metrics: a review. Talanta. 2022 Feb 1;238(2):123046. doi: 10.1016/j.talanta.2021.123046, PMID 34801903.