A simple method to quantify lactoferrin in milk by Micro-Batch Resin Extraction (MBRE) before RP-HPLC - Institut Agro Rennes-Angers Accéder directement au contenu
Poster De Conférence Année : 2017

A simple method to quantify lactoferrin in milk by Micro-Batch Resin Extraction (MBRE) before RP-HPLC

Résumé

Comparison with immunological methods Separative techniques (HPLC, capillary or SDS-PAGE) were rarely used for the quantification of lactoferrin (Lf) in milk compared to immunochemical methods (ELISA, RID, biosensing) which were found more specific and sensitive, but also more expensive and moderately reproducible. We present here a new HPLC-based method validated for the quantification of lactoferrin in bovine milk. Lactoferrin, due to its cationic character (pI > 8), is positively charged at the pH of native milk (6,8-7) whereas almost all caseins and whey proteins (pI < 5,6) are negatively charged. This difference is used to specifically and quantitatively adsorb Lf on a strong cationexchange resin assay developed at a micro-batch scale. Lf is then desorbed increasing ionic strength without any dilution as compared to its concentration in milk. Finally, Lf is fully separated from other co-extracted minor cationic substances and quantified by conventional RP-HPLC with UV detection. The extraction using resin replaced the acid precipitation of caseins which was used in most HPLC-based methods causing loss of lactoferrin and keeping whey proteins in sample extract that interfered with Lf in HPLC. Lf Adsorption Rinses Lf Desorption RP-HPLC Weigh 100 mg resin 1 + 1 mL skim milk 2 60 rev.min-1 , 90 min, RT 3
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Dates et versions

hal-03678911 , version 1 (25-05-2022)

Identifiants

  • HAL Id : hal-03678911 , version 1

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Sylvie Pochet, Céline Arnould, Perrine Poton, Jocelyne Guinard-Flament, Eric Beuvier. A simple method to quantify lactoferrin in milk by Micro-Batch Resin Extraction (MBRE) before RP-HPLC. XIIIth International Conference on Lactoferrin: Structure, Function and Application, Nov 2017, Rome, Italy. ⟨hal-03678911⟩
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