Structure–retention diagrams of ceramides established for their identification

Molecular species analysis of ceramides was carried out using porous graphitic carbon with gradient elution: chloroform–methanol from 45:55 to 85:15 with a slope at 2.7%/min. These conditions gave a linear relationship between retention data and structure of ceramides. It was demonstrated that linearity occurred when a high slope value of linear gradient elution was used. Thereby the linear diagram was evolved by plotting the adjusted retention time against the total number of carbon atoms of ceramide molecules. Each line represents one ceramide class. Such a Structure–Retention Diagram describes ceramide retention and thus constitutes an identification method using only retention data. This Structure–Retention Diagram was assessed and compared to another obtained from octadesyl-grafted silica in terms of their reproducibility, precision and ability to provide ceramide identification. Better identification was obtained using the results from both Structure–Retention Diagrams. This approach with a two-dimensional separation system allowed to take advantage of the specificity of both identification models.