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The objective of this investigation was to develop a HPTLC method for the determination of ceftriaxone, cefixime and cefotaxime, cephalosporins widely used in clinical practice. High performance TLC of cephalosporins was performed on pre-coated silica gel HPTLC plates with concentrating zone (2.5x10 cm) by development in mobile phase ethyl acetate-acetone-methanol-water (5:2.5:2.5:1.5 v/v/v/v). A TLC scanner set at 270 nm was used for direct evaluation of the chromatograms in reflectance/absorbance mode. The calibration curves were established as dependence of peak height (linear and polynomial regression) and peak area (polynomial regression) versus ng level (125-500 ng for all cephalosporins investigated). Relative standard deviations obtained from calibration curves was compared. Precision (RSD: 1.12-2.91% (peak height versus ng) and RSD: 1.05-2.75% (peak area versus ng)), and detection limits (ng level) was validated and found to be satisfactory. The method was found to be reproducible and convenient for quantitative analysis of ceftriaxone, cefixime and cefotaxime in their raw materials and their dosage forms. Copyright (C) 1998 Elsevier Science B.V.