[Method development of reversed-phase preparative liquid chromatography for insulin]

Area of Science:

• Biochemistry
• Analytical Chemistry
• Chromatography

Background:

• Insulin purification is crucial for therapeutic applications.
• Developing efficient preparative chromatography methods is essential for isolating biomolecules.

Purpose of the Study:

• To develop and optimize a reversed-phase preparative liquid chromatography method for insulin.
• To investigate chromatographic retention, sample loading, and peak broadening for insulin purification.

Main Methods:

• Optimization of mobile phase gradient and stationary phases.
• Investigation of peak broadening matrix (sample loading, forward/backward broadening).
• Selection of optimal chromatographic system and gradient program.

Main Results:

• Identified optimal stationary phase for narrower peak broadening.
• Achieved focused concentration distribution in the middle of elution.
• Successfully validated by purifying crude insulin products, effectively removing impurities.

Conclusions:

• The developed method provides effective impurity removal for insulin.
• This approach offers practical value and serves as a reference for purifying other macromolecular compounds.