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. 2017:2017:1529280.
doi: 10.1155/2017/1529280. Epub 2017 Jul 11.

Development and Validation of a Stability-Indicating HPLC Method for the Simultaneous Determination of Florfenicol and Flunixin Meglumine Combination in an Injectable Solution

Nidal Batrawi  1 , Hani Naseef  1 , Fuad Al-Rimawi  2
Affiliations

Development and Validation of a Stability-Indicating HPLC Method for the Simultaneous Determination of Florfenicol and Flunixin Meglumine Combination in an Injectable Solution

Nidal Batrawi et al. J Anal Methods Chem. 2017.

Abstract

The combination of the powerful antimicrobial agent florfenicol and the nonsteroidal anti-inflammatory flunixin meglumine is used for the treatment of bovine respiratory disease (BRD) and control of BRD-associated pyrexia, in beef and nonlactating dairy cattle. This study describes the development and validation of an HPLC-UV method for the simultaneous determination of florfenicol and flunixin, in an injectable preparation with a mixture of excipients. The proposed RP-HPLC method was developed by a reversed phase- (RP-) C18e (250 mm × 4.6 mm, 5 μm) column at room temperature, with an isocratic mobile phase of acetonitrile and water mixture, and pH was adjusted to 2.8 using diluted phosphoric acid, a flow rate of 1.0 mL/min, and ultraviolet detection at 268 nm. The stability-indicating method was developed by exposing the drugs to stress conditions of acid and base hydrolysis, oxidation, photodegradation, and thermal degradation; the obtained degraded products were successfully separated from the APIs. This method was validated in accordance with FDA and ICH guidelines and showed excellent linearity, accuracy, precision, specificity, robustness, LOD, LOQ, and system suitability results within the acceptance criteria.

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Figures

Figure 1
Figure 1
Chemical structure of florfenicol and flunixin meglumine.
Figure 2
Figure 2
Chromatogram of Flr and Flx in drug product using the developed method in this study.
Figure 3
Figure 3
Chromatogram of stress testing of Flr and Flx under basic hydrolysis condition of 0.02 N NaOH, at RT for 2 hours.
Figure 4
Figure 4
Chromatogram of stress testing of Flr and Flx under acidic hydrolysis condition of 1 N HCl, at 40°C for 2 days.
Figure 5
Figure 5
Chromatogram of stress testing of Flr and Flx under oxidative condition of 0.2% H2O2 at 40°C, protected from light for 7 days.
Figure 6
Figure 6
Chromatogram of Flr and Flx under thermal stress testing condition of 75°C for 14 days.
See this image and copyright information in PMC

References

    1. United States Pharmacopeial Convention. Vet. Syst. Florfenicol. United States Pharmacopeial Convention, p. 6, 2007.
    1. British Pharmacopoeia. Monograph on Flunixin meglumine. British Pharmacopoeia. 2013 doi: 10.1111/j.1365-2885.2000.00284.x. - DOI
    1. United States Pharmacopeial Convention. Vet. Syst. Flunixin. United States Pharmacopeial Convention, p. 5, 2007.
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