Quick bloanalytical method of quantification and validation of sulbactam in plasma using high performance liquid chromatography

A quick method of quantitative determination of sulbactam in human plasma, using liquid chromatography-UV spectroscopy, has been developed and validated. After derivatization with imidazole, plasma samples were treated by direct deproteinization with acetonitrile as an extraction solvent. After ultracentrifugation, sulbactam extract was directly injected onto the LC column. Chromatographic separation was performed on TSK Gel Super ODS (50 mm x 4.6 mm i.d., 2 mu m) using methanol and phosphate buffer with tetrabutylammonium hydroxide solution as a mobile phase. Gradient elution was employed. The method was fully validated according to the United States Food and Drug Administration requirements (linearity, precision, trueness, quantification limit, detection limit, recovery, specificity and stability), The calibration curves were linear within the concentration range of 0.05-4.0 mu g mL(-1). Good method/system precision and accuracy of the method were demonstrated.