A simple micro-batch ion-exchange resin extraction method coupled with reverse-phase HPLC (MBRE-HPLC) to quantify lactoferrin in raw and heat-treated bovine milk - Institut Agro Rennes-Angers Accéder directement au contenu
Article Dans Une Revue Food Chemistry Année : 2018

A simple micro-batch ion-exchange resin extraction method coupled with reverse-phase HPLC (MBRE-HPLC) to quantify lactoferrin in raw and heat-treated bovine milk

Résumé

Lactoferrin is an iron-binding cationic glycoprotein (pI = 8.7) beneficial for mammal health, especially udder and milk preservation. A new simple two-step method of quantification was developed. Lactoferrin in 1 mL of bovine skim milk was first adsorbed onto 100 mg of macroporous sulfonated-resin at pH 6.8 by rotary stirring for 90 min at 20–25 °C. After washing the resin, lactoferrin was desorbed using 1 mL of 2 M NaCl containing phenylalanine as a dilution marker, then fully resolved and quantified by RP-HPLC at 220 nm using a wide-bore C4 silica column. This robust, inexpensive and flexible method improves selectivity (no protein interference) and sensitivity compared to previous HPLC methods. In-laboratory validation demonstrated its linearity (25 to 514 µg Lf mL(−1)), accuracy (110 to 98% recovery), and precision (< 4%), which were comparable to immunobased methods. The results for individual raw cow’s milk were strongly correlated with results using an ELISA test.
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Dates et versions

hal-01770347 , version 1 (18-04-2018)

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Citer

Sylvie Pochet, Céline Arnould, Perrine Poton, Jocelyne Guinard-Flament, Odile Rolet-Répécaud, et al.. A simple micro-batch ion-exchange resin extraction method coupled with reverse-phase HPLC (MBRE-HPLC) to quantify lactoferrin in raw and heat-treated bovine milk. Food Chemistry, 2018, 259, pp.36-45. ⟨10.1016/j.foodchem.2018.03.058⟩. ⟨hal-01770347⟩
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