Separation of Lipid Classes by HPLC on a Cyanopropyl Column

A new method for the separation and identification of lipid classes by normal-phase HPLC on a cyanopropyl column is described. The use of a simple binary gradient, with toluene as a component, provided a rapid separation of non-polar as well as phospholipid classes. The inherent small differences in performances between possible non-polar eluent components of the gradient, such as hexane, heptane, and iso-octane, had a pronounced impact on retention times for individual phospholipid classes. Separation of molecular species within a lipid class could also be observed.