DEVELOPMENT AND VALIDATION OF A GREEN BIOANALYTICAL METHOD FOR THE DETERMINATION OF SPARFLOXACIN IN PHARMACEUTICAL DOSAGE FORM AND HUMAN PLASMA BY RP-HPLC

Abstract

This study describes the development of an innovative, green, rapid, precise, selective and sensitive reverse phase high-performance liquid chromatography method for the quantitative determination of Sparfloxacin (SPR) in human plasma and pharmaceutical dosage form. Extraction of drug from plasma was done by employing optimized liquid-liquid extraction procedure. The sample was analyzed using Methanol: Water (pH-7 with triethylamine) (60:40 % v/v) as mobile phase. Chromatographic separation was achieved on Prontosil C-18 column (4.6 x 250mm, 5μ particle size) as stationary phase using isocratic elution (at a flow rate of 1 ml/min). The peak was detected using UV-PDA detector set at 254 nm and the total time for a chromatographic separation was 9 min. The calibration curve obtained was linear (r2= 0.9998) over the concentration range of 5-25μg/ml. Method was validated for precision, robustness and recovery. The limit of detection (LOD) and limit of quantitation (LOQ) was 0.573 and 1.542 μg/ml respectively. There was no significant difference between the amount of drug spiked in plasma and the amount recovered and plasma did not interfere in estimation. Other important factors is that the method uses less amounts of organic solvent, produces low levels of waste and does not use buffer solution, minimizing effluent treatment, which contributes to the environment and implements methods aimed green chemistry, making economic for the industry. Thus, the proposed method is suitable for the analysis of SPR in tablet dosage forms and human plasma.