Reversed-phase high-performance liquid chromatography of insulin and insulin derivatives : A comparative study

The reversed-phase separation of crystalline insuline (I) and monoiodoinsulins (II) has been investigated, with respect to the effects of buffer, substitution group, pore-size and column support backbone. The separations were performed either isocratically (for II) or by gradient elution with very n...

Full description

Saved in:
Bibliographic Details
Published in:Journal of Chromatography A 1985, Vol.348 (2), p.347-361
Main Authors: Welinder, Benny S., Linde, Susanne, Hansen, Bruno
Format: Article
Language:English
Subjects:
Analysis
Biological and medical sciences
General pharmacology
high-performance liquid chromatography
insulin
Medical sciences
Pharmacology. Drug treatments
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The reversed-phase separation of crystalline insuline (I) and monoiodoinsulins (II) has been investigated, with respect to the effects of buffer, substitution group, pore-size and column support backbone. The separations were performed either isocratically (for II) or by gradient elution with very narrow gradients (for I). Fourteen reversed-phase columns, the majority being silica-based, were investigated, and three main result emerged. (1) Trifluoroacetic acid is unsuitable as a buffer for this type of analysis, whereas trialkylammonium phosphates are very suitable. (2) The separation between the major components in crystalline insulin was comparable in the main for all the columns tested except one. However, the ability to distinguish between the numerous minor components (co-extracted with insulin peptide) varied a great deal between the columns. (3) In an optimized buffer system only three columns were able to separate insulin peptide and the four monoiodoinsulin isomers; all three were 80—100-Å silica-based C 18 columns.
ISSN:0021-9673
DOI:10.1016/S0021-9673(01)92473-2