Stability Evaluation on Diminazene Diaceturate and Phenazone in Bulk and Combined Formulations using Validated Chromatographic Method
Abstract
The combined therapy of diminazene aceturate (DMZ) and phenazone (PHENZ) is widely used in veterinary medicine to combat trypanosomiasis and babesiosis. This study presents a novel, validated HPLC method for accurately quantifying DMZ and PHENZ in various pharmaceutical formulations, including bulk powders, sachets, vials, and injectables. The chromatographic separation was achieved using a C18 column (150 x 4.6 mm, 5 μm particle size) with a mobile phase composed of phosphate buffer (pH 3.0) and methanol (70:30 v/v) at a flow rate of 1 mL/minute. UV detection was set at 250 nm. The method demonstrated linearity over a concentration range of 20-100 μg/mL for DMZ and 25-125 μg/mL for PHENZ, with correlation coefficients exceeding 0.999. Forced degradation studies were conducted under various stress conditions to assess the method's stability-indicating power. DMZ exhibited first-order degradation under acidic pH conditions. While slight degradation (2.4-3.1%) was observed under alkaline, UV, and indoor room light conditions, PHENZ remained stable. The validated HPLC method effectively quantified DMZ and PHENZ in the presence of their degradation products and impurities, demonstrating its suitability for quality control and stability studies of these combined drug formulations.
Full text article
References
2. Dayoub M, Shnaigat S, Tarawneh RA, Al-Yacoub AN, Al-Barakeh F, Al-Najjar K. Enhancing Animal Production through Smart Agriculture: Possibilities, Hurdles, Resolutions, and Advantages. Ruminants. 2024;4(1):22-46. DOI: 10.3390/ruminants4010003
3. Akode RM, Shantier SW, Gadkariem EA, Mohamed MA. Simultaneous Determination and Stability Studies on Diminazene Diaceturate and Phenazone Using Developed Derivative Spectrophotometric Method. Int J Anal Chem. 2017;2017:4269587. DOI: 10.1155/2017/4269587; PMCID: PMC5299212; PMID: 28246529
4. Kassaye L, Hymete A, Bekhit AA, Genete G. Validation of an HPLC method for the simultaneous determination of diminazene diaceturate and phenazone in injectable veterinary granules and bulk powders. Pak J Pharm Sci. 2012;25(1):255-9. PMID: 22186338
5. Roybal JE, Walker CC, Pfenning AP, Turnipseed SB, Storey JM, Gonzales SA, et al. Concurrent determination of four fluoroquinolones in catfish, shrimp, and salmon by liquidchromatography with fluorescence detection. J AOAC Int. 2002;85(6):1293–301. PMID: 12477191
6. Gilbert RJ. Studies in rabbits on the disposition and trypanocidal activity of the anti-trypanosomal drug, diminazene aceturate (Berenil). Br J Pharmacol. 1983;80(1):133-9. DOI: 10.1111/j.1476-5381.1983.tb11058.x; PMCID: PMC2044970; PMID: 6652366
7. Gummow B, du Preez JL, Swan GE. Paired-ion extraction and high-performance liquid chromatographic determination of diminazene in cattle plasma: a modified method . Onderstepoort J Vet Res. 1995;62(1):1–4. PMID: 8539029
8. Fouda HG. Determination of diminazene in plasma by high-performance liquid chromatography. J Chromatogr Sci. 1977;15(11):537-8. DOI: 10.1093/chromsci/15.11.537; PMID: 925118
9. Turnipseed SB, Clark SB, Andersen WC, Karbiwnyk CM, Miller KE, Hurlbut JA. Confirmation of diminazene diaceturate in bovine plasma using electrospray liquid chromatography-mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2006;844(1):127–33. DOI: 10.1016/j.jchromb.2006.07.016; PMID: 16891161
10. Fouda HG. Gas chromatography chemical ionization mass spectrometric analysis of diminazene in plasma. Biomed Mass Spectrom. 1978;5(1):72–5. DOI: 10.1002/bms.1200050113; PMID: 623896
11. Staroverov SA, Vasilenko OA, Gabalov KP, Pristensky DV, Yermilov DN, Aksinenko NM, et al. Preparation of polyclonal antibodies to diminazene and its detection in animal blood plasma. Int Immunopharmacol. 2008;8(10):1418-22. DOI: 10.1016/j.intimp.2008.05.014; PMID: 18573353
12. Karanja WM, Mdachi RE, Murilla GA. A competetive enzyme-linked immunosorbent assay for diminazene. Acta Trop. 2002;84(2):75-81. DOI: 10.1016/s0001-706x(02)00184-5; PMID: 12429424
13. Campbell M, Prankerd RJ, Davie AS, Charman WN. Degradation of berenil (diminazene aceturate) in acidic aqueous solution. J Pharm Pharmacol. 2004;56(10):1327–32. DOI: 10.1211/0022357044409; PMID: 15482649
14. Abualhasan MN, Batrawi N, Zaid AN, Watson DG. A validated and stability indicating HPLC method for analysis of diminazene aceturate and antipyrine combination in a ready injectable solution. Drug Res. 2013;63(6):300-4. DOI: 10.1055/s-0033-1337939; PMID: 23532624
15. Atsriku C, Watson DG, Tettey JN, Grant MH, Skellern GG. Determination of diminazene aceturate in pharmaceutical formulations by HPLC and identification of related substances by LC/MS. J Pharm Biomed Anal. 2002;30(4):979-86. DOI: 10.1016/s0731-7085(02)00450-8; PMID: 12408888
16. Miao HF, Cao M, Xu DY, Ren HY, Zhao MX, Huang ZX, et al. Degradation of phenazone in aqueous solution with ozone: influencing factors and degradation pathways. Chemosphere. 2015;119:326-33. DOI: 10.1016/j.chemosphere.2014.06.082; PMID: 25038548
17. The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. Validation of Analytical Procedures: Text and Methodology Q2(R1). In: ICH Harmonised Tripartite Guideline. Geneva: The International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; 2005. 1-13.
18. Ermer J, Kleinschmidt G, Rignall A, McGregor P, Agut C. Method Performance Qualification. In: Ermer J, Nethercote P, editors. Method Validation in Pharmaceutical Analysis: A Guide to Best Practice, Second, Completely Revised and Updated Edition. Method Validation in Pharmaceutical Analysis: A Guide to Best Practice, Second, Completely Revised and Updated Edition. Weinheim: Wiley-VCH Verlag; 2014. DOI: 10.1002/9783527672202.ch7
19. Guimaraes GJ, Bartlett MG. The critical role of mobile phase pH in the performance of oligonucleotide ion-pair liquid chromatography-mass spectrometry methods. Future Sci OA. 2021;7(10):FSO753. DOI: 10.2144/fsoa-2021-0084; PMCID: PMC8610006; PMID: 34840810
20. Blessy M, Patel RD, Prajapati PN, Agrawal YK. Development of forced degradation and stability indicating studies of drugs-A review. J Pharm Anal. 2014;4(3):159-65. DOI: 10.1016/j.jpha.2013.09.003; PMCID: PMC5761119; PMID: 29403878
21. Teja GS, Archana D, Srinu B, Ali SKA, Reddy SSN, Parvez SK, et al. A Comprehensive Guide for Analytical Method Validation. Int J Pharm Sci Rev Res. 2023;82(2):5-17. DOI: 10.47583/ijpsrr.2023.v82i02.002
22. Borman P, Elder D. Q2(R1) Validation of Analytical Procedures. In: Teasdale A, Elder D, Nims RW. ICH Quality Guidelines: An Implementation Guide. New Jersey: John Wiley & Sons; 2017. DOI: 10.1002/9781118971147.ch5
23. González-González O, Ramirez IO, Ramirez BI, O'Connell P, Ballesteros MP, Torrado JJ, et al. Drug Stability: ICH versus Accelerated Predictive Stability Studies. Pharmaceutics. 2022;14(11):2324. DOI: 10.3390/pharmaceutics14112324; PMCID: PMC9693625; PMID: 36365143
24. Kako D, Ghareeb MM, Al-Lami MS. High-Performance Liquid Chromatography (HPLC) Method Validation for Identifying and Quantifying Rebamipide in Ethosomes. Cureus. 2024;16(3):e56061. DOI: 10.7759/cureus.56061; PMCID: PMC11009477; PMID: 38618434
25. Jenke JR. Chromatographic Method Validation: A Review of Current Practices and Procedures. I. General Concepts and Guidelines. J Liq Chromatogr Relat Technol. 1996;19(5):719-36. DOI: 10.1080/10826079608005533
26. Shantier SW, Gadkariem EA, Adam MO, Mohamed MA. Development of stability-indicating methods for cefquinome sulphate. Int J Biomed Sci. 2013;9(3):162-7. PMCID: PMC3809348; PMID: 24170991
27. Wehr CT. Sample preparation and column regeneration in biopolymer separations. J Chromatogr. 1987;418:27-50. PMID: 3305543
28. Horvath C, Melander W, Molnar I, Molnar P. Enhancement of retention by ion-pair formation in liquid chromatography with nonpolar stationary phases. Anal Chem. 1977;49(14):2295-305. DOI: 10.1021/ac50022a048
29. Food and Agriculture Organization of the United Nations. Residues of some veterinary drugs in animals and foods. Rome: Joint FAO/WHO Expert Committee on Food Additives; 2004.
Authors
Copyright (c) 2024 Amna Mubarak, Shaza Wagiealla Shantier, Magdi Awadalla Mohamed, Elrasheed Ahmed Gadkariem
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Authors continue to retain the copyright to the article if the article is published in the Borneo Journal of Pharmacy. They will also retain the publishing rights to the article without any restrictions.
Authors who publish in this journal agree to the following terms:
- Any article on the copyright is retained by the author(s).
- The author grants the journal the right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share work with an acknowledgment of the work authors and initial publications in this journal.
- Authors can enter into separate, additional contractual arrangements for the non-exclusive distribution of published articles (e.g., post-institutional repository) or publish them in a book, with acknowledgment of their initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their websites) prior to and during the submission process. This can lead to productive exchanges and earlier and greater citations of published work.
- The article and any associated published material are distributed under the Creative Commons Attribution-ShareAlike 4.0 International License.