Analysis of insulins by high-performance liquid chromatography. II. Separation of various species of insulins.

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Abstract
A reversed-phase high-performance liquid chromatographic (RP-HPLC) method for the separation of bovine, porcine, ovine and equine insulins has been investigated. The complete chromatographic separation of these four insulins was achieved on a column of LiChrosorb RP-18 (5μm) with a mixture of acetonitrile and 5mM tartrate buffer (pH 3.0) containing sodium sulfate as the mobile phase at 40°C. The capacity factors of the four insulins were greatly affected by the concentrations of acetonitrile and sodium sulfate in the mobile phase and by the column temperature. Careful preparation of the mobile phase and a constant column temperature are essential for the reproducible and complete separation of insulins by this RP-HPLC method.

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