Optimization of fat-soluble vitamin separation by supercritical fluid chromatography

Summary Fat-soluble vitamin separation, achievable using a micropacked SFC column loaded with Cw 20M, is optimized using a rotatable central composite experimental design. Two chromatographic response functions, based on relative retention and the number of peaks resolved are proposed. Temperature and pressure gradients giving the best response in the experimental region are obtained and the effect of their variation on separation is evaluated. Relative standard deviations resulting from both absolute and normalized peak areas are also given.