PURPOSE: To perform the accurate analysis of lipid within a short time, by a method wherein a fraction soluble in hot methanol and n-hexane is collected from a blood specimen to be injected into a reciprocal column to elute an objective lipid and, after elution is finished, the supply of a moving phase is stopped to send an org. solvent-containing washing liquid.

CONSTITUTION: After a blood specimen is dripped in hot methanol to remove serum protein, n-hexane is added to said specimen to collect a fraction soluble in hot methanol and n-hexane. This fraction is applied to a reciprocal column by using a moving phase prepared by adjusting the pH of a water/methanol mixed liquid to 2.4 with sodium phosphate and phosphoric acid and, for example, the peak P1 of testosterone is detected by using ultraviolet rays with a wavelength of 210nm. At this time, the supply of the moving phase is stopped and a washing liquid prepared by mixing an equal amount of isopropanol and water is sent to elute unknown components P2', P3' and, in this stage, the washing liquid remaining in the column is washed off by again sending the moving phase and, after large unknown component P4 is detected, the supply of liquid is succeeded until a base line is fixed. By this method, analysis is enabled without inhibiting the next analysis with unknown components.