The application of reverse phase high-performance liquid chromatography to the separation and analysis of cephradine and cephalexin is demonstrated. The procedure has been applied to chemicals, pharmaceutical formulations and reaction solutions. The preparation of samples is simple and rapid. Chromatographic conditions are described for both pellicular and small particle columns. The feasibility of determining cephradine and cephalexin in physiological fluids has also been demonstrated.Cephradine and cephalexin are members of the cephalosporin family, an important new class of antibiotics. As shown in Fig. 1, these compounds have a 13-lactam and a fused six-member sulfurcontaining ring and differ by only one double bond. Cephalexin contains the phenylglycine moiety attached to the nitrogen at the 7-position, while in the case of cephradine the dihydrophenylglycine moiety is attached.Both cephradine and cephalexin, as well as other cephalosporin antibiotics, are frequently determined by microbiological assay or chemical methods, such as iodometric titrations, hydroxylamine assay or the ninhydrin reaction1). These chemical methods are based on functional groups common to both compounds and do not distinguish between cephradine and cephalexin. However, the highperformance liquid chromatographic techniques presented here provide more accurate, convenient, rapid and specific methods of analysis for these compounds. In a previous paper2), we described the application of reverse phase high-performance liquid chromatography to many cephalosporins and other antibiotics. Varian used a variation of our approach for the separation of 7-ADCA and cephalexin using a small particle column3). This paper presents the application of reverse phase chromatography to the separation and quantitation of cephradine and cephalexin in various media.
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