Skip to Main content Skip to Navigation
Journal articles

Size-exclusion HPLC as a sensitive and calibrationless method for complex peptide mixtures quantification

Abstract : This work describes an original methodology to quantify complex peptide mixtures by size-exclusion high-performance liquid chromatography (SE-HPLC). The methodology was first tested on simulated elutions of peptide mixtures. For this set of experiments, a good estimation of the total peptide concentration was observed (error less than 10%). Then 30 fractions obtained by ultrafiltration of hydrolysates from two different sources were titrated by Kjeldahl or BCA analysis and analysed by SE-HPLC for an experimental validation of the methodology. Very good matchs between methods were obtained. The linear working range depends on the hydrolysate but is generally between 0.2 and 4g L-1 (i.e. between 10 and 200 mu g). Moreover, the presence of organic solvents or salts in samples does not impact the accuracy of the methodology contrary to common quantification methods. Hence, the findings of this study show that total concentration of complex peptide mixture can be efficiently determinate by the proposed methodology using simple SE-HPLC analysis.
Complete list of metadatas

https://hal.univ-lorraine.fr/hal-01264857
Contributor : Lrgp Ul <>
Submitted on : Friday, January 29, 2016 - 5:06:14 PM
Last modification on : Friday, March 20, 2020 - 3:54:15 PM

Identifiers

Collections

Citation

Alice Bodin, Xavier Framboisier, Dominique Alonso, Ivan Marc, Romain Kapel. Size-exclusion HPLC as a sensitive and calibrationless method for complex peptide mixtures quantification. Journal of Chromatography B - Analytical Technologies in the Biomedical and Life Sciences, Elsevier, 2015, 1006, pp.71-79. ⟨10.1016/j.jchromb.2015.09.035⟩. ⟨hal-01264857⟩

Share

Metrics

Record views

382