Simultaneous HPLC Assay of Gliclazide and Ciprofloxacin in Plasma and its Implementation for Pharmacokinetic Study in Rats

Abstract

Abstract A simple and reliable high-performance liquid chromatography method for simultaneous quantitation of gliclazide and ciprofloxacin in plasma sample has been developed and validated. This method implements protein precipitation, a simple and practical pretreatment method by the addition of acetonitrile that gives a clean supernatant. The separation was carried out in a system consisted of a C18 column with acetonitrile and KH2PO4 (0.01 M, 0.1% v/v of triethylamine, pH 2.7) as the mobile phase in a gradient elution at a total flow-rate of 1 mL/min. Gliclazide and ciprofloxacin were quantitated using an ultraviolet detector set at wavelengths of 229 and 277 nm, respectively, which ensures optimal sensitivity for both compounds. This method possesses an excellent linearity at concentration ranges of 0.5–50 mg/L for gliclazide and 0.1–10 mg/L for ciprofloxacin. High within- and between-run accuracy for both gliclazide (% error of −8.00 to 0.45%) and ciprofloxacin (% error of −10.00 to 7.63%) were demonstrated. The intra- and inter-day precision (expressed as %CV) was <8 and 12% for gliclazide and ciprofloxacin, respectively. Both analytes were stable during storage and sample processing. The method reported in this study can be implemented for pharmacokinetic interaction study in rats.