Green validated chromatographic methods for simultaneous determination of co-formulated oxytetracycline HCl and flunixin meglumine in the presence of their impurities in different matrices

Abstract

Green analytical chemistry is an important area of chemical sciences. So, developing competent eco-friendly analytical tools is a big challenge. This study devotes two chromatographic techniques for the simultaneous determination of a binary formulated mixture of oxytetracycline HCl (OXY) and flunixin meglumine (FLU), and their impurities tetracycline (TRC) and 2-chloronicotinic acid (CNA), respectively. Primarily, a TLC densitometric method is proposed and validated using TLC plates sprayed with 5% EDTA of pH 9 as a stationary phase and (acetonitrile/ distilled water/ ethanol 7:2:1, by volume) as a developing system. Well-separated spots are detected at 267 nm where linear relations have been achieved at 0.05–2 μg/band and 0.01–2 μg/band for OXY and FLU, respectively. The second developed and validated method is HPLC which is fulfilled on Hypersil BDS column-C18. A mobile phase of distilled water with 0.1%TFA/ acetonitrile in a ratio of 82: 18, v/v firstly then 70: 30, v/v after 9 min is used in a sequential isocratic elution at 210 nm for FLU and 267 nm for OXY, TRC, and CNA. Sensitive and wide linearity ranges are achieved at 0.05–200 μg/mL for both drugs. The two methods are applied successfully in cattle meat and milk for cited drugs determination. According to ICH guidelines, a validation study has been accomplished for the proposed methods. Statistical comparison has been carried out with official and reported methods. Eventually, the greenness of both procedures is evaluated using Eco-Scale which gives eco-friendly results.Graphical abstract