ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF SODIUM PHENYLBUTYRATE AND TAURURSODIOL BY USING RP-HPLC IN BULK AND PHARMACEUTICAL DOSAGE FORM
DOI:
https://doi.org/10.22159/ijap.2026v18i1.55081Keywords:
Sodium phenylbutyrate, Taurursodiol, RP-HPLC, Development, ValidationAbstract
Objective: In order to assess Sodium Phenylbutyrate and Taurursodiol, two active pharmaceutical components, simultaneously, this study set out to create and validate a new RP-HPLC (Reverse Phase High Performance Liquid Chromatography) technique that was simple, responsive, and stable.
Methods: An X-bridge phenyl column with dimensions 150x4.6 mm and a 3.5 µ was used in the chromatographic approach. The mobile phase consisted of acetonitrile and 0.1% triethyl amine with a pH of 2.5, adjusted with OPA (Ortho Phosphoric Acid) in a ratio of 40:60. Isocratic elution was used. The instrumental conditions were set to 1.0 ml/min with a detection wavelength of 266 nm using the PDA (Photo Diode Array) detector. Validation of the proposed method was carried out according to an international conference on harmonization (ICH) guidelines.
Results: The calibration curves were linear, with a regression coefficient (R2) value of 0.999 and concentrations ranging from 75 to 450 µg/ml of Sodium phenylbutyrate and 25–150 µg/ml of Taurursodiol. The method’s LOD (Limit Of Detection) and LOQ (Limit Of Quantification) were 0.63 µg/ml, 0.21 µg/ml, and 2.1 µg/ml, 0.7 µg/ml for Sodium Phenylbutyrate and Taurursodiol, respectively.
Conclusion: The proposed method to be fast, simple, feasible and affordable in assay condition. During stability tests, it can be used for routine analysis of production samples and to verify the quality of drug samples during stability studies.
References
1. Merritt JL, Brody LL, Pino G, Rinaldo P. Newborn screening for proximal urea cycle disorders: current evidence supporting recommendations for newborn screening. Mol Genet Metab. 2018;124(2):109-13. doi: 10.1016/j.ymgme.2018.04.006, PMID 29703588.
2. Milazzo G, Mercatelli D, Di Muzio G, Triboli L, De Rosa P, Perini G. Histone deacetylases (HDACs): evolution, specificity role in transcriptional complexes and pharmacological actionability. Genes. 2020;11(5):556. doi: 10.3390/genes11050556, PMID 32429325.
3. Shahbazi J, Liu PY, Atmadibrata B, Bradner JE, Marshall GM, Lock RB. The bromodomain inhibitor JQ1 and the histone deacetylase inhibitor panobinostat synergistically reduce N-Myc expression and induce anticancer effects. Clin Cancer Res. 2016;22(10):2534-44. doi: 10.1158/1078-0432.CCR-15-1666, PMID 26733615.
4. Shteinberg M, Haq IJ, Polineni D, Davies JC. Cystic fibrosis. Lancet. 2021;397(10290):2195-211. doi: 10.1016/s0140-6736(20)32542-3, PMID 34090606.
5. Xu X, Zhang X, Zhang G, Abbasi Tadi D. Prevalence of antibiotic resistance of Staphylococcus aureus in cystic fibrosis infection: a systematic review and meta-analysis. J Glob Antimicrob Resist. 2024;36:419-25. doi: 10.1016/j.jgar.2023.05.006, PMID 37211214.
6. Motta AP, Rigobello MC, Silveira RC, Gimenes FR. Nasogastric/nasoenteric tube-related adverse events: an integrative review. Rev Lat Am Enfermagem. 2021;29:e3400. doi: 10.1590/1518-8345.3355.3400, PMID 33439952.
7. Kodua M, Mackenzie JM, Smyth N. Nursing assistants experiences of administering manual restraint for compulsory nasogastric feeding of young persons with anorexia nervosa. Int J Ment Health Nurs. 2020;29(6):1181-91. doi: 10.1111/inm.12758, PMID 32578949.
8. Arghya M, Santosh S. Andrographolide prevents acute hyperammonemia-induced motor dysfunction in rats. Evidences for its mechanism of action. Phytomedicine Plus. 2025;5(3):100679. doi: 10.1016/j.phyplu.2024.100679.
9. McGlacken Byrne SM, Conway GS. Premature ovarian insufficiency. Best Pract Res Clin Obstet Gynaecol. 2021;81:98-110. doi: 10.1016/j.bpobgyn.2021.09.011, PMID 34924261.
10. Seppa S, Kuiri Hanninen T, Holopainen E, Voutilainen R. Management of endocrine disease: diagnosis and management of primary amenorrhea and female delayed puberty. Eur J Endocrinol. 2021;184(6):R225-42. doi: 10.1530/EJE-20-1487, PMID 33687345.
11. Bernstein H, Bernstein C. Bile acids as carcinogens in the colon and at other sites in the gastrointestinal system. Exp Biol Med (Maywood). 2023;248(1):79-89. doi: 10.1177/15353702221131858, PMID 36408538.
12. Masato A, Plotegher N, Boassa D, Bubacco L. Impaired dopamine metabolism in Parkinson’s disease pathogenesis. Mol Neurodegener. 2019;14(1):35. doi: 10.1186/s13024-019-0332-6, PMID 31488222.
13. Martinez M, Harding CO, Schwank G, Thony B. State-of-the-art 2023 on gene therapy for phenylketonuria. J Inherit Metab Dis. 2024 Jan;47(1):80-92. doi: 10.1002/jimd.12651, PMID 37401651.
14. Ashe K, Kelso W, Farrand S, Panetta J, Fazio T, De Jong G. Psychiatric and cognitive aspects of phenylketonuria: the limitations of diet and promise of new treatments. Front Psychiatry. 2019;10:561. doi: 10.3389/fpsyt.2019.00561, PMID 31551819.
15. Chiang JY, Ferrell JM. Bile acid receptors FXR and TGR5 signaling in fatty liver diseases and therapy. Am J Physiol Gastrointest Liver Physiol. 2020;318(3):G554-73. doi: 10.1152/ajpgi.00223.2019, PMID 31984784.
16. Shapiro H, Kolodziejczyk AA, Halstuch D, Elinav E. Bile acids in glucose metabolism in health and disease. J Exp Med. 2018;215(2):383-96. doi: 10.1084/jem.20171965, PMID 29339445.
17. Petersen KS, Kris Etherton PM. Diet quality assessment and the relationship between diet quality and cardiovascular disease risk. Nutrients. 2021;13(12):4305. doi: 10.3390/nu13124305, PMID 34959857.
18. Blood Pressure Lowering Treatment Trialists’ Collaboration. Pharmacological blood pressure lowering for primary and secondary prevention of cardiovascular disease across different levels of blood pressure: an individual participant-level data meta-analysis. Lancet. 2021;397(10285):1625-36. doi: 10.1016/S0140-6736(21)00590-0, PMID 33933205.
19. Handsaker RE, Kashin S, Reed NM, Tan S, Lee WS, McDonald TM. Long somatic DNA-repeat expansion drives neurodegeneration in Huntington’s disease. Cell. 2025;188(3):623-639.e19. doi: 10.1016/j.cell.2024.11.038, PMID 39824182.
20. McColgan P, Tabrizi SJ. Huntington’s disease: a clinical review. Eur J Neurol. 2018;25(1):24-34. doi: 10.1111/ene.13413, PMID 28817209.
21. Alborghetti M, Nicoletti F. Different generations of type-B monoamine oxidase inhibitors in Parkinson’s disease: from bench to bedside. Curr Neuropharmacol. 2019;17(9):861-73. doi: 10.2174/1570159X16666180830100754, PMID 30160213.
22. Abusrair AH, Elsekaily W, Bohlega S. Tremor in Parkinson’s disease: from pathophysiology to advanced therapies. Tremor Other Hyperkinet Mov (NY). 2022;12:29. doi: 10.5334/tohm.712, PMID 36211804.
23. Masrori P, Van Damme P. Amyotrophic lateral sclerosis: a clinical review. Eur J Neurol. 2020;27(10):1918-29. doi: 10.1111/ene.14393, PMID 32526057.
24. Vitturi BK, Montecucco A, Rahmani A, Dini G, Durando P. Occupational risk factors for multiple sclerosis: a systematic review with meta-analysis. Front Public Health. 2023;11:1285103. doi: 10.3389/fpubh.2023.1285103, PMID 38054069.
25. Tateishi H, Monde K, Anraku K, Koga R, Hayashi Y, Ciftci HI. A clue to unprecedented strategy to HIV eradication: “Lock-in and apoptosis”. Sci Rep. 2017;7(1):8957. doi: 10.1038/s41598-017-09129-w, PMID 28827668.
26. Raju C, Sivagami B. Validation of a stability-indicating analytical method development for simultaneous estimation of sodium phenylbutyrate and taurursodiol in bulk and formulation using UPLC method. Iran J Pharm Sci. 2025;21(1):70-81. doi: 10.22037/ijps.v21i1.45528.
27. Beludari MI, Chedela HK, Lingareddy SS, Dara CB, Karimulla S. Development and validation of a stability-indicating analytical method for simultaneous determination of sodium phenylbutyrate and taurursodiol in bulk and formulation using reverse phase ultra-performance liquid chromatography. Egypt Pharm J. 2024;23(2):264-71. doi: 10.4103/epj.epj_295_23.
28. Malyaadri M, NL Janaki. Analytical method development and validation of sodium phenyl butyrate and taurursodiol by using RP-UPLC in bulk and pharmaceutical dosage form. High Technol Lett. 2024;30:164-77.
29. Karishma SK, Anusha B, Vegesna S. Stability indicating method devleopment and validation for the estimation of sodium phenyl butyrate and taurursodilol by using RP-HPLC. Afr J Biomed Res. 2024;27(4S):15422-32. doi: 10.53555/AJBR.v27i4S.7699.
30. Bhavani P, Prasada Rao K, Mohan S. Novel validated reversed-phase high-performance liquid chromatography method for determination of glucosamine diacerein and methyl sulfonyl methane in micro sample rat plasma and its application to pharmacokinetic and dissolution studies. Asian J Pharm Clin Res. 2020;13(12):50-63. doi: 10.22159/ajpcr.2020.v13i12.36547.
31. Gomathy S, Narenderan ST, Meyyanathan SN, Gowramma B. Development and validation of HPLC method for the simultaneous estimation of apigenin and luteolin in commercial formulation. J Crit Rev. 2020;7(19):4785-90. doi: 10.31838/jcr.07.19.560.
32. Supriya T, Naresh D, Vijaya Kumar G, Haneer MA. Stability indicating RP-HPLC method development and validation for simultaneous estimation of escitalopram and flupentixol pure and marketed formulation. Asian J Pharm Res. 2018;8:4-10.
33. Naykode MD, Bhagwat DA, Jadhav SD, More HN. Analytical and bioanalytical method for quantification of pure azilsartan not its salts by RP-HPLC. Res J Pharm Technol. 2017;10(3):708-14. doi: 10.5958/0974-360X.2017.00133.0.
34. Singh M, Charde M, Shukla R, Rita MC. Determination of calcipotriene in calcipotriene cream 0.05% w/w by RP-HPLC method development and validation. Res J Pharm Technol. 2011;4(8):1219-23.
35. Kumar SA, Debnath A, Rao JV, Sankar DG. Development and validation of a sensitive RP-HPLC method for simultaneous estimation of rosuvastatin and fenofibrate in tablet dosage form by using PDA detector in Gradient mode. Res J Pharm Technol. 2016;9(5):549-54. doi: 10.5958/0974-360X.2016.00104.9.
36. Manoranjani M. A study of method development validation and forced degradation for simultaneous quantification of cisplatin and fluorouracil in bulk and pharmaceutical dosage form by RP-HPLC. J Pharm Sci Res. 2021;13:155-61.
37. Prasada Rao PT. A study of method development and validation for simultaneous estimation of pemetrexed and cisplatin using RP-HPLC. J Pharm Sci Res. 2021;13:143-8.
38. Satyadev TN. A new selective separation method development and validation of cabozantinib and Nivolumab using HPLC. J Pharm Sci Res. 2021;13:188-92.
39. David Raju M. Development and validation of HPLC method for the determination of lasmiditan drug in bulk and tablet dosage form. J Pharm Sci Res. 2021;13(3):170-3.
40. Baje Syed IB, Nannapaneni M. Bioanalytical validation method for Capmatinib and Spartalizumab in rabbit plasma by using highly effective mass spectrophotometric method. Rasayan J Chem. 2022;15(4):2748-55. doi: 10.31788/RJC.2022.1547098.
41. Shivani CP, Maheshwari DG. Development and validation of UV spectrometric and HPLC method for estimation of escitalopram oxalate and flupentixol dihydrochloride in combined dosage form. AJPTI. 2016;4(17):59-70.
42. Shanmugasundaram P, Kamarapu SK. RP-HPLC method for the simultaneous estimation and validation of amlodipine besylate and atenolol in bulk and tablet dosage form in biorelevant dissolution medium (Fassif). Res J Pharm Technol. 2017;10(10):3379-85. doi: 10.5958/0974-360X.2017.00601.1.
43. ICH harmonized tripartite guideline. Validation of analytical procedures: text and methodology Q2(R1) International conference on the harmonization; 2005.
44. Raziq A, Jan SU. Relative comparison of stability and degradation of methylcobalamin tablets of different brands at different storage settings. Int J Appl Pharm. 2021;13:171-75.
45. Rafi S, Rambabu K. Stability indicating validated HPLC method for the determination of aceclofenac and misoprostol in bulk and pharmaceutical formulation. IJRPS. 2020;11(4):7848-53. doi: 10.26452/ijrps.v11i4.4669.
46. Swati K, Abhishek P, Sushank S, Bothiraja C, Atmaram P. High-performance liquid chromatography for the simultaneous estimation of cefoperazone and sulbactam in rat plasma and its importance in therapeutic drug monitoring. Int J Pharm Pharm Sci. 2020;12(10):92-7. doi: 10.22159/ijpps.2020v12i10.38638.
47. Vejendla A, Talari S, Moturu R, Murthy Boddapati SN, Kola AE. Method development and validation for Cabotegravir and Rilpivirine by using HPLC and its degradants are characterized by LCMS and FTIR. Futur J PharmSci. 2021;7:1-18. doi: 10.1186/s43094 021 00355 8.
48. Charu Pandya P, Sadhana Rajput J. Development and validation of stability indicating method RP-HPLC method of acotiamide. Int J Pharm Pharm Sci. 2018;10(9):1-8. doi: 10.22159/ijpps.2018v10i9.24925.
49. Athavia BA, Dedania ZR, Dedania RR, Swamy SM, Prajapati CB. Stability indicating HPLC method for determination of vilazodone hydrochloride. Int J Curr Pharm Sci. 2017;9(4):123-9. doi: 10.22159/ijcpr.2017v9i4.20975.
Published
How to Cite
Issue
Section
Copyright (c) 2026 SUNIL RAYUDU, M. MANORANJANI, D. ANITHA
This work is licensed under a Creative Commons Attribution 4.0 International License.