Development and Validation of a High Performance Liquid Chromatography Ultraviolet Detection Method for the Quantitative Determination of Vancomycin Hydrochloride

Vancomycin hydrochloride is a tricyclic glycopeptide that contains amino acids and sugars. This substance is indicated to treat serious infections caused by Gram-positive bacteria by intravenous infusion. The objective of this study was to develop and validate an analytical methodology by high performance liquid chromatography with ultraviolet detection (HPLC-UV) to determine vancomycin hydrochloride content by assessing the parameters of selectivity, linearity, working range, matrix effect, robustness, precision, and accuracy. The sample used was vancomycin hydrochloride in a vial and analyzes were carried out on HPLC-UV system, with C18 reverse-phase column at 30 °C, pH=4 and diode-array detection (220 nm). The mobile phase was composed of acetonitrile and monobasic ammonium phosphate buffer (8:92 v/v), 1 mL min-1 flow, injection volume of 20 μL and 15 minute of run time. The method has been shown to be selective, free from mobile phase interference, diluent and other substances on vancomycin hydrochloride retention time; the method is linear in the range between 25 and 175 µg mL-1; matrix effect showed parallelism between the lines, thus indicating the absence of interference of the matrix constituents in analysis of the compound of interest; the method was robust with drug variations proportional to the deliberate changes caused by the change in the flow rate of the mobile phase and in the column temperature; the method showed accuracy at 25, 50, and 75 µg mL-1 concentrations, showing satisfactory recovery rate after addition of the standard. The analytical methodology described proved to be simple, fast, safe and was considered valid.