Low molecular weight collagen peptides have gained a lot of interest as it has the potential to stimulate bone and cartilage regeneration, prevent skin aging, and other health benefits. Although collagen has been long used for medical and cosmetic purposes, a standard efficient analysis technique to determine the molecular weight and distribution of collagen has not yet been developed. Gel permeation chromatography (GPC) is an analytical technique that can measure the molecular weight and generate information about the structure, conformation as well as distribution of polymers. Collagen is made hydrophilic through various enzyme treatments for efficacy when ingested and analyzed using an aqueous column. However, due to the presence of a small number of carboxyl groups in the packing material of aqueous columns, an ionic interaction occurs resulting in interference. To minimize and overcome this barrier, salts and other additives can be added to the mobile phase when analyzing collagen. Therefore, in this study, an attempt was made to develop a multi-angle approach to improve the distribution analysis of collagen peptides. GPC analysis conditions were optimized for different salts and additive in order to reduce the interference and successful results were obtained.