Accuracy and Precision Comparison for Molecular Weight Distribution Assay of Fish Collagen Peptides: a Methodology Study Between Two Gel Permeation Chromatography Columns

摘要

Two gel permeation chromatography columns, Superdex Peptide HR 10/30 column and TSK G2000 SWXL column, were compared for their accuracy and precision for determining the molecular weight distribution of fish collagen peptides using methodology study. To confirm the results, collagen peptides were subjected to ultrafiltration separation and then the filtrates (MW&3kDa) were loaded to MALDI-TOF-MS. Superdex column was consider as the most promising candidate for the determination of small molecular weight peptides (MW&3kDa), since it spiked recovery was 101.90% and the RSD was below 1%. As for TSK column, it was more suitable for the assay of large molecular weight peptides (MW&10kDa). The spiked recovery was 105.66% and the RSD was below 0.5%. Besides, acetonitrile reagent narrowed the pore size distribution of column packing materials, which made it more inaccurate for the analysis of smaller molecular weight peptides.