Determination of true molar masses of collagen peptides

Collagen peptides are relatively small proteins with typical molar masses of less than 10.000 g/mol. They exhibit a molar mass distribution, MMD, which influences their properties.

A monograph by GME describes a procedure how to determine true MMDs for collagen peptides using standard Liquid Chromatography, LC, equipment (1). The LC setup comprises an isocratic pump, an injection system, GPC/SEC columns and an UV/VIS detector.

Figure 1: Step 2 of the guided setup processing the overlay of chromatograms of the five collagen peptide reference samples with assigned weight average molar masses

Column calibration is a two-step procedure. A base calibration is established using one CNBr-Peptides sample. Applying broad standard calibration using 5 different broadly distributed collagen peptide reference materials allows transforming the calibration curve into a chemically matching collagen peptide calibration curve to obtain true molar masses (2).

PSS WinGPC UniChrom software offers a convenient wizard for broad calibration, which guides the users through each step of the calibration and automatically adjusts the base calibration curve such that best agreement with target values is obtained. The resulting calibration curve allows for the determination of true molar masses of collagen peptides.

Read the full application note here

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