Use of principal component analysis for investigation of factors affectingretention behaviour of ceramides on porous graphitized carbon column

The potentiality of a porous graphitized carbon stationary phase was assess ed for ceramide analysis in nonaqueous reversed phase chromatography with a n evaporative light scattering detection. The influence of the nature of th e mobile phase was first performed under isoeluotropic isocratic conditions . Eight mobile phase compositions in the presence or absence of triethylami ne/formic acid were tested with a set of structurally relevant ceramides. V alues obtained under these conditions for the peak heights, retention facto rs, plate numbers. asymmetry factors, relative retentions and Time-Correcte d Calibrated Normalised Resolution Product were analysed by Principal Compo nent Analysis. Principal Component Analysis was effective in attributing re lationships between the chromatographic behaviour and analytical conditions . The relationship between the increase of the detector response and the pr esence of triethylamine/formic acid has been confirmed, and the main change of selectivity has been attributed to interactions provided by the weak so lvent of the mobile phase. These conclusions have allowed the development o f gradient conditions for the analysis of complex ceramide mixtures on poro us graphitized carbon column.