DEVELOPMENT AND VALIDATION OF ANALYTICAL METHODS FOR IMMUNOSUPPRESSIVE AGENT USING UV- SPECTROPHOTOMETRY AND RP-HPLC.
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Keywords
UV Spectrophotometry, RP-HPLC, Immunosuppressive Agent, Method Development, Method Validation, Pharmaceutical Analysis, Tacrolimus. Dosage Form. Bulk Analysis, Analytical Chemistry
Abstract
This dissertation focuses on the development and validation of analytical methods for the quantification of immunosuppressive agents, specifically Tacrolimus, using UV spectrophotometry and reverse-phase high-performance liquid chromatography (RP-HPLC) in both bulk and dosage forms. A simple and rapid UV-Vis spectrophotometric method for the estimation of Tacrolimus was developed, with calibration curves constructed and the linearity range determined. Additionally, a precise, accurate, and robust HPLC method for the quantification of Tacrolimus in bulk and pharmaceutical formulations was established, optimizing chromatographic conditions including mobile phase composition, flow rate, detection wavelength, and column selection. The HPLC method was validated according to ICH guidelines, demonstrating excellent specificity, linearity, accuracy, precision, LOD, LOQ, and robustness. Comparative analysis between the UV-Vis spectrophotometric and HPLC methods highlighted the former as a viable alternative for Tacrolimus quantification. These validated methods support routine quality control and stability studies, ensuring regulatory compliance, and facilitating the development of innovative drug delivery systems. They also contribute to the understanding of Tacrolimus's physicochemical properties, ultimately enhancing its therapeutic management.
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References
1. Harris DC. Exploring Chemical Analysis. 5th ed. Harris DC, editor. New York: W.H. Freeman and Company; 2012 p. 1–25, 101–115. 2. Christian GD. Analytical Chemistry: John Wiley & Sons; 2004 p. 1-748. 3. Braun RD. Introduction to Instrumental Analysis: McGraw-Hill; 1987 p. 197, 202-205, 205-210, 311-318. 4. Christian GD. Analytical Chemistry. 6th ed.: John Wiley & Sons; 2004 p. 355–360. 5. Kellner R, Mermet JM, Otto M, Widmer HM. Analytical Chemistry: A Modern Approach to Analytical Science. 2nd ed.: John Wiley & Sons; 2004 p. 240–245. 6. Harris DC. Quantitative Chemical Analysis: W. H. Freeman; 2010. 7. Kellner R, Mermet JM, Otto M, Widmer HM. Analytical Chemistry: A Modern Approach to Analytical Science: John Wiley & Sons; 2004 p. 1-1300. 8. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). Validation of Analytical Procedures: Text and Methodology Q2(R1). 2005;: 1-17. 9. Michalski R. Environmental Analysis. Critical Reviews in Analytical Chemistry. 2011; 41: 35-58. 10. Gossner CM, Schlundt J, Ben Embarek P, Hird S, Lo-Fo-Wong D, Beltran JJO, et al. The melamine incident: Implications for international food and feed safety. Environmental Health Perspectives. 2009; 117: 1803-1808. 11. Skoog DA, Holler FJ, Crouch SR. Principles of Instrumental Analysis. 7th ed.: Cengage Learning; 2017 p. 325, 331, 339-344, 344-347, 835-842. 12. Silverstein RM, Webster FX, Kiemle DJ. Spectrometric Identification of Organic Compounds. 8th ed.: Wiley; 2014 p. 143, 145, 156-160, 160-163, 423-425. 13. Sharma BK. Instrumental Methods of Chemical Analysis. 24th ed.: Goel Publishing House; 2007 p. 212, 220-225, 225-230, 310-315. 14. Rahman A, Aziz A, Ahmad M. Application of UV-Vis Spectrophotometry in Environmental Analysis. Journal of Environmental Science. 2020. 15. Kumar R, Singh P, Mehta R. Quality Control in the Food Industry Using UV-Visible Spectrophotometry. Food Chem. 2018. 16. Johnston A, Smith B, Patel N. Clinical Applications of UV-Vis Spectrophotometry in Diagnostics. Clinical Biochemistry. 2021. 17. Gupta S, Sharma R, Verma P. Applications of HPLC in Clinical Diagnostics. Clinical Biochemistry. 2019. 18. Gupta S, Sharma R, Verma P. Kinetics of Enzyme-Catalyzed Reactions Using UV-Vis Spectrophotometry. Biochemical Journal. 2019. 19. United States Pharmacopeia. USP General Chapter : Validation of Compendial Procedures. 2019 November.. 20. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Harmonised Guideline: Validation of Analytical Procedures Q2(R2). 2023 November. Final Version, Adopted on 1 November 2023. 21. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Harmonised Tripartite Guideline: Pharmaceutical Quality System Q10. 2008 June.. 22. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Harmonised Guideline: Analytical Procedure Development Q14. 2023 November. Final Version, Adopted on 1 November 2023. 23. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Harmonised Tripartite Guideline: Pharmaceutical Development Q8(R2). 2009 August.. 24. Zhou M, Veenstra TD. Mass Spectrometry: m/z 1983–2008. Biotechniques. 2008; 44: 659–668. 25. Willard HH, Merritt LL, Dean JA, Settle FA. Instrumental Methods of Analysis. 7th ed.: CBS Publishers and Distributors; 1988 p. 118-148. 26. Welch CJ, Wu N, Biba M, Hartman R, Brkovic T, Gong X, et al. Greening analytical chromatography. TrAC Trends in Analytical Chemistry. 2010 September; 29: 667–680. 27. Weidner SM. Mass Spectrometry: MALDI (Matrix-Assisted Laser Desorption/Ionization) and ESI (Electrospray Ionization). Polymer Science: A comprehensive reference. 2012;: 259–280. 28. Wang X, Guo Z, Zhang D, Yan Y, Yu Y, Du B, et al. Integrating liquid chromatography-electrochemical detection-surface Enhanced Raman Spectroscopy on microfluidic chip for phenylurea herbicides analysis. Sens. Actuators, B. 2024; 407: 135436. 29. Rathee P, Rathee S, Thakur S, Kumar V. Simultaneous estimation of Amlodipine besylate and Atenolol as API and in tablet dosage forms by vierodt’s method using UV spectrophotometry. International Journal of Current Trends in Research. 2010; 2: 62-68. 30. Rashmin M. An introduction to analytical method development for pharmaceutical formulations. Pharmainfo.net. 2008; 6: 1-11. 31. Rahman A, Aziz A, Ahmad M. Application of HPLC in Environmental Analysis. Journal of Environmental Science. 2020. 32. Prathap B, Rao GHS, Devdass G, Dey A, Harikrishnan N. Review on Stability Indicating HPLC Method Development. International Journal of Innovative Pharmaceutical Research. 2012; 3: 229-237. 33. Porfireva A, Goida A, Evtugyn V, Evtugyn G. Impedimetric sensor based on molecularly imprinted polythionine from deep eutectic solvent for epinephrine determination. Green Analytical Chemistry. 2024; 4: 100040. 34. Pavia DL, Lampman GM, Kriz GS. Introduction to Spectroscopy. 3rd ed.: Thomson Learning; 2001 p. 356. 35. Ozaki Y. Near-infrared spectroscopy—Its versatility in analytical chemistry. Anal. Sci. 2012; 28: 545–562. 36. Niu Y, Chen Y, Zhou J, Sun W. Online electrochemistry coupling liquid chromatography-mass spectrometry for rapid investigation on the phase I and phase II simulated metabolic reactions of pharmaceuticals. Anal. Bioanal. Chem. 2024 March; 416(1): 53–62. 37. Kumar R, Singh P, Mehta R. Quality Control in the Food Industry Using HPLC. Food Chemistry. 2018. 38. Khopkar SM. Basic Concepts of Analytical Chemistry. 2nd ed. New Delhi: New Age International Publishers; 1998 p. 45–78, 150–175. 39. Ke J, Gao C, Folgueiras-Amador AA, Jolley KE, de Frutos O, Mateos C, et al. Self-Optimization of Continuous Flow Electrochemical Synthesis Using Fourier Transform Infrared Spectroscopy and Gas Chromatography. Appl. Spectrosc. 2021 December; 76: 38–50. 40. Kayasth S, Swain K. Role of analytical chemistry in environmental monitoring. J. Radioanal. Nucl. Ch. 2004; 262: 35–39. 41. Kasture AV, Mahadik KR, Wadodkar SG, More HN. Pharmaceutical Analysis. 7th ed.: Nirali Publication; 2007 p. 28-30. 42. Johnston A, Smith B, Patel N. HPLC in Biochemical Research. Biochemical Journal. 2021. 43. Jeffery GH, Bassett J, Mendham J, Denney RC. Vogel's Textbook of Quantitative Chemical Analysis. 5th ed.: Longman Scientific and Technical Publishers; 1991 p. 3-13. 44. Inturi R, Raju MD, Rao MVB, Inturi S. Validation of LC-MS/MS Method for Monitoring of Upadacitinib in Rat Plasma Samples: A review. Institutional Narasaraopeta of Pharmaceutical Sciences. 2022;: 102-106. 45. Horváth C, Nahum A, Frenz JH. High-performance displacement chromatography. J. Chromatogr. A. 1981; 203: 219–230. 46. Harris DC, Lucy CA. Quantitative Chemical Analysis. 10th ed.: W.H. Freeman & Company; 2020 p. 1-911. 47. Gossner CM, Schlundt J, Embarek PK, Hird S, Lo-Fo-Wong D, Beltran JJO, et al. Food Safety: Contaminants and Toxins. Infectious Disease Clinics. 2009; 23: 973-993. 48. Gonçalves-Filho D, De Souza D. Detection of synthetic antioxidants: what factors affect the efficiency in the chromatographic analysis and in the electrochemical analysis? Molecules. 2022; 27(20): 7137. 49. Giselbrecht S, Länge K, Huang TJ, Manz A. Revisiting lab-on-a-chip technology for drug discovery. Nat. Rev. Drug Discov. 2012; 11: 371–387. 50. Dole MN, Patel PA, Sawant SD. Advance applications of Fourier transform infrared spectroscopy. International Journal of Pharmaceutical Sciences Review and Research. 2011. 51. Dittrich PS, Manz A. Lab-on-a-chip: microfluidics in drug discovery. Nat. Rev. Drug Discov. 2006; 5: 210– 218. 52. Beebe P, Tambe V, Dhole S, Patil V. An update on various analytical techniques based on UV spectroscopy used in determination of dissociation constant. International Journal of Pharmaceutical Sciences Review and Research. 2014; 4: 278-285. 53. Beckett AH, Stenlake JB. Practical Pharmaceutical Chemistry. 4th ed.: CBS Publishers and Distributors; 2004 p. 284-300, 162-163. 54. United State Pharmacopeia. Validation of Compendial Procedures: United State Pharmacopeia; 2010. 55. United States Pharmacopeia. USP General Chapter : Analytical Procedure Lifecycle. 2020 November.. 56. International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. ICH Harmonised Tripartite Guideline: Quality Risk Management Q9. 2005 November.. 57. U.S. Food and Drug Administration. Guidance for Industry: Analytical Procedures and Methods Validation for Drugs and Biologics. 2015 July.. 58. PubChem. Tacrolimus. 2024. Available from: \url 59. Organization WH. Tacrolimus: International Pharmacopoeia. 2024. Available from: \url 60. Kino T, Hatanaka H, Miyata S, Inamura N, Nishiyama M, Yajima T, et al. FK-506, a novel immunosuppressant isolated from a Streptomyces. I. Fermentation, isolation, and physico-chemical and biological characteristics. The Journal of antibiotics. 1987; 40: 1249–1255. 61. Kelly PA, Burckart GJ, Venkataramanan R. Tacrolimus: a new immunosuppressive agent. American Journal of Health-System Pharmacy. 1995; 52: 1521–1535. 62. Food US, Administration D. Tacrolimus. 2024. Available from: \url 63. Faulds D, Goa KL, Benfield P. Tacrolimus. A further update of its pharmacology and therapeutic use in the management of organ transplantation. Drugs. 1993; 46: 441–484. 64. DrugBank. Tacrolimus. 2024. Available from: \url 65. Agency EM. Tacrolimus: EPAR - Product Information. 2024. Available from: \url
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DECEMBER 27, 2024
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