Collagen peptides (also referred to as hydrolysed collagen or collagen hydrolysate) are produced from natural proteins by hydrolysis. They are soluble in cold water, are highly digestible and used in dietary supplements and functional foods.
Collagen peptides are relatively small proteins with typical molar masses of less than 10.000 g/mol comprising at least two and at most 100 amino acids. Common to nearly all macromolecules, collagen peptides do not exhibit only one specific molar mass but a molar mass distribution, MMD. This MMD influences their properties.
GPC/SEC is an established technique to measure MMDs of macromolecules, such as collagen peptides, but suffers from 2 limitations:
- It is a relative technique which allows determination of close to true molar masses only
- if reference materials chemically matching the analyte structure are available
- or if advanced detectors such as online viscometers or online light scattering detectors are applied
- It is a liquid chromatographic technique which requires highly qualified and trained users, especially when advanced detectors such as online viscometers or online light scattering detectors are employed.
The purpose of the present work was to develop a simple, cost-effective and robust high precision GPC/SEC method for QC, allowing to provide close to true molar masses for collagen peptides. The method should be applicable in any laboratory allowing for reliable inter laboratory comparison. Therefore, the use of molar mass sensitive detectors should be avoided.
The single steps in the study were
- identification of collagen peptides suitable as reference materials and determination of their characteristics
- establishing a suitable calibration procedure
- round robin test of the final approach
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