Drug Res (Stuttg) 2013; 63(06): 300-304
DOI: 10.1055/s-0033-1337939
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

A Validated and Stability Indicating HPLC Method for Analysis of Diminazene Aceturate and Antipyrine Combination in a Ready Injectable Solution

M. N. Abualhasan
1   Department of Pharmacy, An-Najah National University, Nablus, Palestine
N. Batrawi
2   Advanced Veterinary Company, Ramallah, Palestine
A. N. Zaid
1   Department of Pharmacy, An-Najah National University, Nablus, Palestine
D. G. Watson
3   Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK
› Institutsangaben
Weitere Informationen


received 21. Dezember 2012

accepted 17. Februar 2013

26. März 2013 (online)


Diminazene aceturate and Antipyrine combination therapy is widely used in veterinary medicine. A simple reverse HPLC method for the analysis of samples of a ready injectable formulation containing a mixture of active ingredients and inactive excipients has been developed. The HPLC analysis was carried out using a reversed phase (RP)-C18 (250 mm×4.0 mm, 5 μm) column. The isocratic mobile phase consisted of a mixture of acetonitrile, methanol, phosphate buffer and hexane sulfonate; the flow rate was 0.6 mL/min and ultraviolet detection was at 291 nm. This method was validated in accordance with FDA and ICH guidelines and showed good linearity, accuracy, precision, selectivity and the system suitability results were within the acceptance criteria. A stability-indicating study was also carried out and indicated that this method could be used for purity and degradation evaluation of these formulations.

  • References

  • 1 Turnipseed SB, Clark SB, Andersen WC et al. Confirmation of diminazene diaceturate in bovine plasma using electrospray liquid chromatography-mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2006; 844: 127-133
  • 2 Schad GJ, Allanson A, Mackay SP et al. Development and validation of an improved HPLC method for the control of potentially counterfeit isometamidium products. J Pharm Biomed Anal 2008; 46: 45-51
  • 3 http://www.merckmanuals.com/vet/circulatory_system/blood_parasites/babesiosis.html Accessed: 03.12.2012
  • 4 Martindale . Reynolds JEF, Parfitt K, Parsons AV, Sweetman SC. eds. The Extra Pharmacopoeia. The Pharmaceutical Press; London: 1989: 34-35
  • 5 Merck Index . An encyclopedia of chemicals, drugs and biologicals. Merck and Co., Inc; USA: 1989: 113
  • 6 Macharia JC, Bourdichon AJ, Gicheru MM. Efficacy of Trypan: a diminazene based drug as antileishmanial agent. Acta tropica 2004; 92: 267-272
  • 7 Boothe DM, Cullen JM, Calvin JA et al. Antipyrine and caffeine dispositions in clinically normal dogs and dogs with progressive liver disease. Am J Vet Res 1994; 55: 254-261
  • 8 http://www.alibaba.com/showroom/dd-and-antipyrine-granules.html) http://mercatorpharma.com/products/734/veterinary%20products/dd-antipyrine-injection Accessed: 01.12.2013
  • 9 http://mercatorpharma.com/products/734/veterinary-products/c7/diminazene-antipyrine-injection Accessed: 05.12.2013
  • 10 Kassaye L, Hymete A, Bekhit AA et al. 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: 255-259
  • 11 Alvi NK, Haqqi TM, Hadi SM. Irreversible binding of berenil, a trypanocidal drug to blood proteins. Indian J Exp Biol 1985; 23: 172-173
  • 12 Aliu YO, Mamman M, Peregrine AS. Pharmacokinetics of diminazene in female Boran (Bos indicus) cattle. J Vet Pharmacol Ther 1993; 16: 291-300
  • 13 Atsriku C, Watson DG, Tettey JN et al. Determination of diminazene aceturate in pharmaceutical formulations by HPLC and identification of related substances by LC/MS. J Pharm Biomed Anal 2002; 30: 979-986
  • 14 Eichelbaum M, Sonntag B, Dengler HJ. HPLC determination of antipyrine metabolites. Pharmacology 1981; 23: 192-202
  • 15 Patil SR, Kumar L, Kohli G et al. Validated HPLC Method for Concurrent Determination of Antipyrine, Carbamazepine, Furosemide and Phenytoin and its Application in Assessment of Drug Permeability through Caco-2 Cell Monolayers. Sci Pharm 2012; 80: 89-100
  • 16 Shargel L, Cheung WM, Yu AB. High-pressure liquid chromatographic analysis of antipyrine in small plasma samples. J Pharm Sci 1979; 68: 1052-1054
  • 17 Shen J, Wanwimolruk S. A Simple and Sensitive HPLC Method for Antipyrine in Plasma. Journal of Liquid Chromatography 1991; 14: 2801-2808
  • 18 Weber A, Opheim K, Smith AL. Simplified High Performance Liquid Chromatographic Quantitation of Antipyrine. Journal of Chromatographic Science 1984; 22: 239-240
  • 19 Ghulam AS. Validation of high-performance liquid chromatography methods for pharmaceutical analysis: Understanding the differences and similarities between validation requirements of the US Food and Drug Administration, the US Pharmacopeia and the International Conference on Harmonization. Journal of Chromatography A 2003; 987: 57-66