Simultaneous Liquid Chromatographic Analysis of the β-Lactam Antibiotics Cefazolin, Cefadroxil, Cephalexin, Ampicillin, and Cephradine in Solution

A liquid chromatographic method was developed for the determination of nanogram quantities of 5 broad-spectrum structurally related beta-lactam antibiotics (cefazolin, cefadroxil, cephalexin, cephradine, and ampicillin) in solution. The method uses a C18 reversed-phase column, UV absorption (240 nm) detection, and an aqueous mobile phase containing isopropyl alcohol and acetic acid. Relative resolution between the antibiotic peaks ranged from 1.7 to 5.9 for all peaks. Chromatographic retention times were 2.97, 3.92, 4.57, 5.37, and 6.56 min for cefazolin, cefadroxil, cephalexin, ampicillin, and cephradine, respectively. Accuracy, precision, linearity, and long term analytical reproducibility were determined by statistical analysis. Use of the proposed method to evaluate the degradation of cephradine solutions stored at room temperature illustrated its potential as a stability-indicating assay.