OBETICHOLIC ACID: AN INSIGHT INTO A QUANTITATIVE DETERMINATION AND METHODOLOGICAL VALIDATION THROUGH NUCLEAR MAGNETIC RESONANCE
Objective: The research work unveils the use of nuclear magnetic resonance (NMR) technique for quantitative determination and method validation of obeticholic acid. As standard expository methodology for more up to date medications or formulations may not be available in pharmacopeias, hence it is fundamental need to create novel analytical procedures which should be precise and accurate.
Methods: Proton (1H) and carbon (13C) NMR analysis were initially performed to confirm the preliminary authenticity of obeticholic acid API. Method validation was accomplished on the basis of standard guidelines for the parameters, in which tetramethylbenzene as an internal standard and deuterated dimethyl sulfoxide as a diluent were used to assess the obeticholic acid.
Results: For the quantification of the drug, the proton nuclear magnetic resonance signals at 0.602 ppm and 6.86 ppm corresponding to the analyte proton of drug and internal standard respectively were used. The curve equation calculated from the regression method, the relative-standard-deviation and correlation-coefficient were found to be 0.743% and 0.9989 respectively, indicating good linearity. Consequently, the quantitative assay of the drug was found to be 99.91% in linearity with limit of detection and quantification values as 0.0773 mg and 0.2344 mg respectively, making successful the study of method validation for obeticholic acid.
Conclusion: The advantage of the method was that no reference standard of analyte drug was required for quantification and method validation. The method is non-destructive and can be applied for quantification of drug in commercial pharmaceutical formulation products.
2. Roberts GC. Applications of NMR in drug discovery. Drug Discovery Today 2000;5:230-40.
3. Van Halbeek H. NMR developments in structural studies of carbohydrates and their complexes. Curr Opin Struc Biol 1994;4:697-709.
4. Gawrisch K, Eldho NV, Polozov IV. Novel NMR tools to study structure and dynamics of biomembranes. Chem Phys Lipids 2002;116:135-51.
5. Holzgrabe U, Deubner R, Schollmayer C, Waibel B. Quantitative NMR spectroscopy-applications in drug analysis. J Pharm Biomed Anal 2005;38:806-12.
6. Wells RJ, Hook JM, Al-Deen TS, Hibbert DB. Quantitative nuclear magnetic resonance (QNMR) spectroscopy for assessing the purity of technical grade agrochemicals: 2, 4-dichlorophenoxyacetic acid (2, 4-D) and sodium 2, 2-dichloropropionate (Dalapon sodium). J Agric Food Chem 2002;50:3366-74.
7. Pellicciari R, Fiorucci S, Camaioni E, Clerici C, Costantino G, Maloney PR, et al. 6?-ethyl-chenodeoxycholic acid (6-ECDCA), a potent and selective FXR agonist endowed with anticholestatic activity. J Med Chem 2002;45:3569-72.
8. Pellicciari R, Costantino G, Camaioni E, Sadeghpour BM, Entrena A, Willson TM, et al. Bile acid derivatives as ligands of the farnesoid X receptor. Synthesis, evaluation, and structure? activity relationship of a series of body and side chain modified analogues of chenodeoxycholic acid. J Med Chem 2004;47:4559-69.
9. Pellicciari R, Costantino G, Fiorucci S. Farnesoid X receptor: from structure to potential clinical applications. J Med Chem 2005;48:5383-403.
10. Markham A, Keam SJ. Obeticholic acid: first global approval. Drugs 2016;76:1221-6.
11. Shah RA, Kowdley KV. Obeticholic acid for the treatment of nonalcoholic steatohepatitis. Expert Rev Gastroenterol Hepatol 2020;14:311-21.
12. Dousa M, Slavíkova M, Kubelka T, Cerny J, Gibala P, Zezula J. HPLC/UV/MS method application for the separation of obeticholic acid and its related compounds in development process and quality control. J Pharm Biomed Anal 2018;149:214-24.
13. Li XY, Zhu SH, Yang F, Hu GX, Yuan LJ. An ultra-performance liquid chromatography-tandem mass spectrometry method for the determination of obeticholic acid in rat plasma and its application in preclinical pharmacokinetic studies. J Chromatogr B 2019;1121:82-8.
14. Edwards JE, LaCerte C, Peyret T, Gosselin NH, Marier JF, Hofmann AF, et al. Modeling and experimental studies of obeticholic acid exposure and the impact of cirrhosis stage. CTS Clin Transl Sci 2016;9:328-36.
15. Olthof PB, Huisman F, Schaap FG, Van LKP, Bennink RJ, Van GRF, et al. Effect of obeticholic acid on liver regeneration following portal vein embolization in an experimental model. Br J Surg 2017;104:590-9.
16. Malz F, Jancke H. Validation of quantitative NMR. Biomed Anal 2005;38:813-23.
17. Harahap Y, Irawan H, Kuswardani. Development and validation of analytical method of 3, 4-methylenedioxy-n-ethylamphetamine in dried blood spot using gas chromatography-mass spec-trometry. Int J Appl Pharm 2020;12:94-9.
18. El-Sheikh R, Hassan WS, Fahmy R, Gouda AA. Validated spectrophotometric methods for determination of montelukast sodium in pure and dosage forms using n-bromosuccinimide and dyes. Int J Appl Pharm 2020;12:152-9.
19. Guideline IH. Validation of analytical procedures: text and methodology Q2 (R1). Vol. 11. InInternational conference on harmonization, Geneva, Switzerland; 2005.
20. Pauli GF, Chen SN, Simmler C, Lankin DC, Go?decke T, Jaki BU, et al. Importance of purity evaluation and the potential of quantitative 1H NMR as a purity assay: miniperspective. J Med Chem 2014;57:9220-31.
21. Bridwell H, Dhingra V, Peckman D, Roark J, Lehman T. Perspectives on method validation: importance of adequate method validation. Qual Assur J 2010;13:72-7.
22. De-Souza SV, Junqueira RG. A procedure to assess linearity by ordinary least squares method. Anal Chimica Acta 2005;552:25-35.
23. Mocak J, Bond AM, Mitchell S, Scollary G. A statistical overview of standard (IUPAC and ACS) and new procedures for determining the limits of detection and quantification: application to voltammetric and stripping techniques (technical report). Pure Appl Chem 1997;69:297-328.
24. Maniara G, Rajamoorthi K, Rajan S, Stockton GW. Method performance and validation for quantitative analysis by 1H and 31P NMR spectroscopy: applications to analytical standards and agricultural chemicals. Anal Chem 1998;70:4921-8.
This work is licensed under a Creative Commons Attribution 4.0 International License.