Validation of quantitation analytical procedure of chloramphenicol in ointment with high performance liquid chromatography (HPLC)

Abstract

     An accurate and sensitive method was developed for the qualitative and quantitative analysis of chloramphenicol in ointment using high performance liquid chromatography (HPLC) with UV detection. Experimental results optimized the chromatographic conditions with the mobile phase component being methanol: dikali hydrophosphat solution at (70:30; v/v) at flow rate of 1.0 mL/min which reduced the running time of the samples and the chromatograms obtained met the symmetry coefficient. At the same time, optimizing the processing of samples with a high fat content,  using a combination of ultrasound and cooling, % recovery efficiency increased significantly from (65.38 to 100.74) % and relative standard deviation %RSD = 0.67 (< 2) compared to ultrasound – assisted extraction method. Quantitative results of chloramphenicol obtained a linear concentration range of (0.09965-0.2989) mg/mL. The accuracy of the method was determined by the recovery rate in the range of (98.49-101.23) %, corresponding to the concentration of (0,1677-0,2392) mg/mL and the repeatability (in %RSD) of 1.05 %. The analytical procedure met the criteria of specificity, linearity, accuracy, and precision set by the guidance of ICH - The International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use. This analytical procedure can be applied in routine quality control of chloramphenicol in ointment.