High pressure liquid chromatographic separation of C18 and C19 Steroids

High Pressure liquid chromatographic procedures for the resolution of mixtures of C18 and C19 steroids of biologic importance are described. On-line monitoring of the absorbance and refractive index of the eluate was found to be suitable for the quantitative measurement of the amounts of eluted steroids. Using a high sensitivity absorbance detector, reliable measurements of as little as 5 ng delta(3)-3 keto steroids were possible.