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Chromatography 2016
September 21-23, 2016
Volume 7, Issue 5(Suppl) J Chromatogr Sep Tech 2016
ISSN: 2157-7064 JCGST, an open access journal
conferenceseries .com
September 21-23, 2016 Amsterdam, Netherlands
World Congress on Chromatography
Size-exclusionHPLCasasensitiveandcalibrationlessmethodforcomplexpeptidemixturesquantification
Romain Kapel, Xavier Framboisier, Dominique Alonso, Ivan Marc and Alice Bodin
University of Lorraine, France
Nowadays, protein hydrolysates and fractions are of great interest because of their nutritional or bioactive properties. The
quantification of total peptide concentration is of a crucial importance in order to establish mass balance of fractionation
processes. This is commonly done either by Kjeldhal analysis, or by colorimetric assays, whose are laborious and time-
consuming. This work describes an original methodology to quantify complex peptide mixtures by size-exclusion high-
performance liquid chromatography (SE-HPLC), already used to characterize the molecular weight distribution of hydrolysate.
In the proposed methodology, each point of the complex mixture chromatogram is regarded as a mixture of peptides sharing
same molar extinction coefficient and molar weight, estimated from its retention time and the hydrolysate aminogram. This
allows a conversion of absorbance into concentration (using Beer-Lambert law) and the integration of the overall signal gives
the peptide concentration of the analysed fraction. The methodology was first tested on simulated elutions of peptide mixtures
and 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 (error less than 15%).
Moreover, the presence of organic solvents or salts in samples does not impact the accuracy of the methodology contrary to
common quantification methods.
Biography
Romain Kapel is a third year PhD student from University of Lorraine, France. In 2013, she graduated as a chemical and process engineer. She is now working on
the fractionation of protein hydrolysate during her PhD program. She will present an original methodology to quantify peptide concentration in complex hydrolysates
by size-exclusion HPLC.
Romain Kapel et al., J Chromatogr Sep Tech 2016, 7:5(Suppl)