The present invention provides a separation process of chromatography for recovering polyunsaturated fatty acid (PUFA) products from a feed mixture comprising: (i) contacting a plurality of aqueous solutions containing an aqueous organic solvent as an eluent to obtain an intermediate product Purifying the feed mixture in a first separation step of a simulated or actual moving bed chromatographic apparatus having a connected chromatotrapy; Obtained in step (i) in a second separation step using a simulated or actual mobile layer chromatography apparatus having a plurality of connected chromatography columns containing an aqueous organic solvent as eluent to obtain the PUFA product, and (ii) Purifying the product; Wherein (a) the first and second separation steps are performed sequentially in the same chromatographic apparatus and the intermediate product is recovered between the first and second separation steps, wherein in the chromatographic apparatus, Adjusted between the first and second separation steps so that the product is separated from the other components of the feed mixture in each separation step; Or (b) the first and second separation steps are performed in separate first and second chromatography devices, the intermediate product obtained from the first separation step is introduced into the second chromatography device, The PUFA product is separated from the other components of the feed mixture in each separation step.
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