High-performance liquid chromatographic separation of citraconylinsulins and preparation of GlyA1PheB1-dicitraconylinsulin.

Abstract
A method for the mutual separation of 6 citraconylated porcine insulins and intact insulin from the reaction mixture of insulin with citraconic anhydride is described, based upon anion-exchange high-performance liquid chromatography on a TSK IEX-540 DEAE column. The method permits the preparative isolation of citraconylinsulins. The optimal reaction conditions for the preparation of GlyA1Phe B1-dicitraconylinsulin, a useful starting material for the preparation of Lys-B29 modified insulin derivatives, were also investigated.

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