DEVELOPMENT AND VALIDATION OF STABILITY INDICATING GREEN HPLC-UV METHOD FOR DETERMINATION OF CEPHALEXIN IN PHARMACEUTICAL DOSAGE FORMS AND HUMAN URINE USING MICELLAR MOBILE PHASE

摘要

Objective : Development and validation of simple, stability indicating and green high performance liquid chromatographic (HPLC) method with ultraviolet (UV) detection for determination of cephalexin in pure form, pharmaceutical dosage forms and human urine samples. Methods: The method is based on using of a micellar mobile phase for separation of cephalexin and its degradation products. The analyte was chromatographed on a Kinetex C 18 75×4.6?mm, 2.6?μm column. Micellar mobile phase composed of 0.1M sodium dodecyl sulphate (SDS) and 10 % isopropanol (IPA), pH was adjusted to 3±0.05 with phosphoric acid, the flow-rate was 1.0 mL/min, the UV detector was set at 254 nm and the injection volume was 20 μl. Stability indicating properties of the proposed method was proved through exposure of the analyte solutions to 4 different stress conditions of acidic, basic, oxidative and photo-irradiation conditions. Results: Under optimized conditions the average recovery was ranged from 100.4–101.7%. The lower limit of quantification (LOQ) and the lower limit of detection (LOD) were 0.097 and 0.029 μg/ml, respectively. A linear correlation in the range of 1–200?μg/ml with the correlation coefficient (r 2 ) of ≥ 0.999 was obtained. Relatively high inter-and intra-day precisions were achieved, the percentage RSD values were lower than 2. The obtained results were validated according to USP validation parameters. Conclusion : The proposed method was found to be not only a greener method but also faster and more convenient than the USP compendial method. Greener here means that the method is more eco-friendly as it avoids usage of toxic solvent and reagent and switch to more benign chemicals. In addition, allow for injection of urine samples directly into an analytical column without pretreatment due to micellar solubilization of the interfering components of the biological samples. Keywords: Green analytical chemistry, Cephalexin, Sodium dodecyl sulfate, Micelle L. C, Human urine sample, Stability indicating method.