Abstract: A chromatographic separation method for separating plural components contained in a liquid to be separated by a chromatography, with a separating device 1 including plural filling portions 10 filled with a separating agent for separating the plural components contained in the liquid to be separated, a supply portion 20 provided in each of the plural filling portions 10 to supply the liquid to be separated or an eluent for extracting any component contained in the liquid to be separated to the filling portion 10, and an extraction portion 30 to extract any component contained in the liquid to be separated from the filling portion 10, the method including: an upward supplying and extracting step of extracting any component contained in the liquid to be separated from an upward stream extraction portion while supplying the eluent to at least one filling portion 10 from an upward stream supply portion by an upward stream.