Citation: PAN Jian, TAO Yunguo. Method development of reversed-phase preparative liquid chromatography for insulin[J]. Chinese Journal of Chromatography, ;2017, 35(8): 848-854. doi: 10.3724/SP.J.1123.2017.04014 shu

Method development of reversed-phase preparative liquid chromatography for insulin

PAN Jian ,
TAO Yunguo

Hisun Pharmaceutical(Hangzhou) Co. Ltd., Hangzhou 311404, China

Received Date: 13 April 2017

The development and optimization of reversed-phase preparative liquid chromatography method for insulin were performed. The chromatographic retention, sample loading and peak broadening were investigated. The mobile phase gradient conditions and stationary phases were optimized. The parameters of the method contained the peak broadening levels under different amounts of sample loading and the concentration distribution of the target compound in the elution curves. The parameters of peak broadening levels were defined and expressed as a matrix, which consisted of sample loading, the forward broadening and the backward broadening levels. The most suitable chromatographic system (including stationary phases and mobile phase conditions) was selected. A gradient program with slow change of the strong elution solvent was used. The most suitable stationary phase should exhibit the narrower peak broadening and the peaks were best to broaden to both sides comparing to that under the analytical conditions. Besides, the concentration distribution of the target compound should be focused on the middle of the elution. The guide principles were validation by purification of crude insulin products. The preparative chromatographic system constructed through this method can remove impurities effectively. The method has good practical value. It can provide reference for the development of preparative chromatographic methods of other macromolecular compounds.
- reversed-phase preparative liquid chromatography,
- chromatographic parameters,
- peak broadening,
- concentration distribution,
- insulin
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