Ion Exchange HPLC of a Marine Collagen

Abstract

HPLC resolution of fibrillar collagen chains, and particularly the α₂ chain from β and γ components, has continued to provide a challenge. A single column method for the clear resolution of the denatured chains of hoki (Macruronus novaezelandiae) skin type I collagen is presented. The effect of changing chromatographic parameters (flow rate, loading, temperature, gradient and solvent, pH) was examined. The α₁ and α₃ chains were readily resolved under all the conditions studied. The α₂ chain was the most difficult component to resolve but also gave the largest response to changes in solvent gradient and pH.

Keywords:

• Collagen
• Hoki
• HPLC

ACKNOWLEDGMENT

This work was carried out under the New Zealand government funding program FRST CO2X0301.

Notes

k' = t₁ − t₀/t₀ where t₀ is the column void time and t₁ is peak retention time.

Rs = 2x (T_R1 − T_R2)/(w₁ + w₂) [15].