Volume : 12, Issue : 06, June – 2025
Title:
DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR ESTIMATION OF SINGLE DRUG IN PHARMACEUTICAL DOSAGE FORM BY RP-HPLC
Authors :
Pooja S. Gonge*, Mahesh S. Armal, Dr. Dhirendra B. Sanghai, Prof. Dr. Nitin S. Bhajipale
Abstract :
The present study focuses on the development and validation of a simple, accurate, and robust reverse-phase high-performance liquid chromatography (RP-HPLC) method for the estimation of Eslicarbazepine Acetate (ESC) in pharmaceutical tablet formulations. ESC, a widely prescribed antiepileptic agent, requires precise quantification to ensure quality control and regulatory compliance. The chromatographic method was optimized using an Inertsil C18 column (4.6 mm × 250 mm, 5 µm) with a mobile phase composed of Acetonitrile and Ammonium Phosphate Buffer (70:30 v/v, pH 3), and detection was performed at 228 nm using a UV detector. The method produced a sharp and well-resolved peak with a retention time of 5.684 minutes and an asymmetry factor of 1.1754, demonstrating excellent efficiency and system suitability. The linearity of the method was confirmed over a concentration range of 80%–120% with a correlation coefficient (R²) of 0.9999. Validation was conducted in accordance with ICH guidelines, assessing parameters such as accuracy, precision, specificity, ruggedness, robustness, and linearity. The method demonstrated high recovery (100.21%) and minimal variability (RSD < 2%), indicating its reliability and reproducibility. The developed RP-HPLC method was successfully applied for the quantitative estimation of ESC in both laboratory mixtures and marketed formulations, showing its potential for routine quality control analysis.
KEYWORDS: Eslicarbazepine Acetate (ESC); RP-HPLC; Method Validation; Pharmaceutical Formulation; Chromatography; Accuracy; Precision; ICH Guidelines
Cite This Article:
Please cite this article in press Pooja S. Gonge et al, Development And Validation Of Analytical Method For Estimation Of Single Drug In Pharmaceutical Dosage Form By Rp-Hplc.,Indo Am. J. P. Sci, 2025; 12(07).
Number of Downloads : 10
References:
1. ICH: Q3A (R2) Impurities in New Drug Substances, Geneva, (2006).
2. Sethi, P. D., High Performance Liquid Chromatography and Quantitative analysis Pharmaceutical Formulations, 1st Edition, CBS Publishers and Distributors, (2001).
3. Jeffery, G. H., Vogel’s textbook of quantitative chemical analysis, 5th Edition, pp- 216-229
4. http://en.wikipedia.org/wiki/Chromatography
5. James, W. Munson., Modern Methods of Pharmaceutical Analysis, Medical Book Distributors, Mumbai, (2001).
6. Erwing, G. W., Instrumental Methods of Chemical Analysis, 2ndEdition, McGraw Hill Book Company, (1960).
7. Sharma, B. K., Instrumental Method of Chemical Analysis, 23rd Edition, Goal Publication House, Meerut, (1991).
8. Sastry, C. S. P., Prasad, T. N. V. and Rao, E. V., Recent application of High Performance Liquid Chromatography in Pharmaceutical Analysis, Indian. Pharm. Education, Vol. 21, (1987).
9. 21 Code of Federal Regulations, Parts 210 and 21, published, (1996).
10. MHRA revises guidelines on “GxP Data Integrity Definitions and Guidance for Industry”, (2016).
11. General Chapter for Physical test: Chromatography Chapter <621>., United States Pharmacopeia, (USP-40 NF-35).
12. Chromatographic separation techniques, European Pharmacopoeia, Vol.7.0, section: 2.2.46, pp. 70-74.
13. Lloyd, R. Snyder., Joseph, J. Kirkland, and joseph. L. Glajch., Practical HPLC Method Development, 2nd Edition, Wiley–Interscience Publication, USA, pp.10-12.
14. Gurdeep, R. Chatwal. and Sham, K. Anand., Instrumental Methods of Chemical Analysis, 5th Edition, Himalaya Publishing House, pp. 1.2 – 1.5, (2002).
15. Sharma, B. K., Instrumental Method of Chemical Analysis, 23rd Edition, Goal Publication House, Merrut, pp: 7-10, (1991).
16. Rama, Rao., Murthy, G. S. S. N. and Khadgapathi, P., High Performance Liquid Chromatography and its role in pharmaceutical analysis, Eastern Pharmacist, Vol. 20, No. 346, (1986).
17. Lloyd, R. Snyder., Joseph, J. Kirkland, and joseph. L. Glajch., Practical HPLC Method Development, 2nd Edition, Wiley–Interscience Publication, USA, pp.359-362.
18. www.fda.gov/cder/guidance/cmc3.pdf.
19. The International Conference on Harmonization (ICH)-Validation of Analytical Procedures: Methodology (Q2B), Food and Drug Administration, USA, (1996).
20. International Conference on Harmonization, “Q2B: Validation of Analytical Procedures: Methodology; Availability,” Federal Register, Vol. 62, No. 96, 27463– 27467, (1997).
21. FDA, “Analytical Procedures and Methods Validation: Chemistry, Manufacturing and Controls Documentation; Availability,” Federal
22. Register (Notices), Vol. 65, No. 169, pp. 52776–52777, (2000).
23. The Merck Index, Merck Research Laboratories, Division of Merck & Co.,14th Edition, Whitehouse Station, NJ, USA, (2006).
24. https://www.drugs.com/avodart.html
25. Yamana, K. and Labrie, F. Luu., The V. Human type 3 5α-reductase is expressed in peripheral tissues at higher levels than types 1 and 2 and its activity is potently inhibited by finasteride and dutasteride. Hormone Molecular Biology and Clinical Investigation, (2010).
26. 5-alpha reductase inhibitors (5-ARIs): Label Change – Increased Risk of Prostate Cancer, U.S. Department of Health & Human Services.
27. www.drugbank.com/drugs/DB06695
28. https://www.pradaxa.com
29. Patel, B., Ram, P., Khatri, T., Ram, V. and Dave, P., Study of Anticoagulant Dabigatran by Analytical Instrumentation, International Letters of Chemistry, Physics and Astronomy, Vol. 11, pp. 233-242, (2014).
30. Prajapati, Kumar, A., Sen S, A. K., Zanwar, A. and Seth, A. K., Spectrophotometric method for estimation of dabigatran etexilate in bulk and its pharmaceutical dosage form, Pharma science monitor, Vol. 5, No. 31, (2014).
31. https://en.wikipedia.org/wiki/Abacavi.
32. Panel on Antiretroviral Guidelines for Adults and Adolescents. Guidelines for the Use of Antiretroviral Agents in HIV-1-Infected Adults and Adolescents. Department of Health and Human Services, USA, (2016).
33. Nachega, J. B., Mugavero, M. J., Zeier, M., Vitoria, M. and Gallant, J. E., Treatment simplification in HIV-infected adults as a strategy to prevent toxicity, improve adherence, quality of life and decrease healthcare costs, Patient Prefer Adherence, Vol: 5, pp. 357–367, (2011)
34. Ford, N. Flexner., C. Vella, S., Ripin, D., and Vitoria, M., Optimization and simplification of antiretroviral therapy for adults and children. CurrOpin HIV AIDS, Vol: 8, pp. S591–S599, (2013).
35. Casado, J. L. and Moreno, S., Potential role of rilpivirine in simplification regimens, Enferm Infecc Microbiol Clin, Vol: 31, pp. S30–S35, (2013).
36. https://www.drugbank.com/drugs/DB06209
37. Prasugrel. Micromedex Drugdex Evaluations online, (2009).
38. European Commission approves Efficient (Prasugrel) for patients with acute coronary syndrome undergoing PCI. Eli Lilly, (2009).
39. https://www.rxlist.com/mozobil-drug.htm
40. Mozobil (plerixafor injection) Full Prescribing Information. Genzyme Corporation, (2013).
41. Lapidot, T. and Petit, I., Current understanding of stem cell mobilization: the roles of chemokines, proteolytic enzymes, adhesion molecules, cytokines, and stromal cells. Exp Hematol, Vol. 30, pp. 973-981, (2002).
42. Martin, C., Bridger, G. J., Rankin, S. M., Structural analogues of AMD3100 mobilise haematopoietic progenitor cells from bone marrow in vivo according to their ability to inhibit CXCL12 binding to CXCR4 in vitro. Br J Amatol, Vol. 134, No. 3, pp. 326-329, (2006).
43. Haresh B. Patel, Rohit H. Dave, Sandip Vadariya Development and Validation of HPLC Method for the Quantification of Impending Genotoxic Impurities in Dapson Drug Substances. Biotechnology Journal International 2024 – Volume 28 ,Issue 4 Page: 1-17
44. Andreas Vrachas , development and validation of a novel RP-HPLC method for the determination of cetrimide and chlorhexidine gluconate in antiseptic solution, Analytica 2022, 3(1), page no 79-91
45. Kumaraswamy Gandla , development and validation of RP-HPLC method for the estimation of lisinoprile in bulk and pharmaceutical dosage form, world journal of pharmaceutical and medical research, Volume 8, issue 8, page no 289-294
46. Samineni R. chimakarthy J, Yamarthy V. Development and validation of analytical method for estimating for balofloxacin in bulk and pharmaceutical dosage form by RP HPLC research general pharmacy and technology 2022 Page no. 2992-2996
47. Baile M Ramole R, and jain A. A Review Analytical Method Development And Validation International Journal of All Research Education & Scientific Methods 2021 page no 450-454
48. M. Mohan Varma, Ashok Thulluru, K. T. Sunil Kumar, G. Sai Kumar and K. Pavani Hplc Method Development And Validation: A Review world journal of pharmaceutical research Vol 10, Issue 11, 2021. Page no 405-426
49. Mohammed Elmowafy Ibrahim El-Bagory Quality by design (QbD) based development and validation of bioanalytical RP-HPLC method for dapagliflozin: Forced degradation and preclinical pharmacokinetic study Journal of Liquid Chromatography & Related Technologies 43(1) page no 1-12
50. V. Kumar, R. Bharadwaj, G.G., S. Kumar, An Overview on HPLC Method Development, Optimization and Validation process for drug analysis, The Pharmaceutical and Chemical Journal, 2(2) (2015) page no 30-40.
51. K. Kardani, N. Gurav, B. Solanki, P. Patel, B. Patel, RP-HPLC Method Development and Validation of Gallic acid in Polyherbal Tablet Formulation, Journal of Applied Pharmaceutical Science. 3 (5) (2013) page no 37-42.
52. Anjali Bakshi, A. Mounika, Shweta Bhutada and Dr. M. Bhagvan Raju Simultaneous Estimation Of Empagliflozin And Linagliptin By RP-HPLC Method world journal of pharmaceutical research Vol 7 Issue 8, 2018. Page no 1062-1071
53. Hani Naseef, Ramzi Moqadi, and Moammal Qurt Development and Validation of an HPLC Method for Determination of Antidiabetic Drug Alogliptin Benzoate in Bulk and Tablets J Anal Methods Chem. 2018; 2018: 1902510.
54. Madhusudan T Bachute, Sunil V Shanbhag, Shankar L Turwale Simultaneous determination of four active pharmaceuticals in tablet dosage form by reversed- phase high performance liquid chromatography Tropical Journal of Pharmaceutical Research October 2019; 18 (10): 2161-2166
55. V. Gupta, A.D. K. Jain, N.S. Gill, K. Gupta, Development and validation of HPLC method – a review, Int. Res J Pharm. App Sci., 2(4) (2012) page no 17-25
56. Mukthi Thammana A Review on High Performance Liquid Chromatography (HPLC) Research & Reviews: Journal of Pharmaceutical Analysis Volume 5 Issue 2 July – September, 2016 page no 22-28