| 摘要 |
PROBLEM TO BE SOLVED: To provide a method that separates a hemoglobin A0 fraction in a short time and with excellent reproducibility, by using a cation exchange chromatography.SOLUTION: A separation method of a hemoglobin A0 fraction by using a cation exchange chromatography, is configured to: use at least two kinds of eluents of an eluent A and an eluent B described below and a column packing agent having a cation exchange group; and set a pressure value generated in a measurement system to 9.8×10Pa or more and 19.6×10Pa or less. (1) The eluent A having a pH value of 4.0 to 6.0 and including all components of (1), (2) and (3) described below, (1) a chaotropic ion, (2) an acid having an acid dissociation constant in a range of 3.5 or more and 6.5 or less and being at least one selected from the group consisting of a monocarboxylic acid, dicarboxylic acid, tricarboxylic acid and amino acid, or a salt thereof, and (3) at least any of inorganic oxo acid and the amino acid having an acid dissociation constant in a range of 6.5 or more and 9.5 or less, or a salt of at least the any. (2) The eluent B having a pH value higher by 0.5 to 3.0 than that of the eluent A, and having a concentration smaller by 20 to 200 mmol/L than that of the eluent A, and to be fed after the eluent A is fed. |
