Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7585119 | Food Chemistry | 2018 | 36 Pages |
Abstract
Lactoferrin is an iron-binding cationic glycoprotein (pIâ¯=â¯8.7) beneficial for mammal health, especially udder and milk preservation. A new simple two-step method of quantification was developed. Lactoferrin in 1â¯mL of bovine skim milk was first adsorbed onto 100â¯mg of macroporous sulfonated-resin at pH 6.8 by rotary stirring for 90â¯min at 20-25â¯Â°C. After washing the resin, lactoferrin was desorbed using 1â¯mL of 2â¯M NaCl containing phenylalanine as a dilution marker, then fully resolved and quantified by RP-HPLC at 220â¯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 514â¯Âµgâ¯Lfâ¯mLâ1), accuracy (110 to 98% recovery), and precision (<4%), which were comparable to immuno-based methods. The results for individual raw cow's milk were strongly correlated with results using an ELISA test.
Keywords
β-LgRP-HPLCPHELOQBSAα-Labovine serum albuminalpha-lactalbuminimmunoglobulinsBeta-lactoglobulinELISAEnzyme-linked immunosorbent assayLOD یا Limit of detectionridRoom temperatureCation-exchange resinSandwich ELISABovine milkCoefficient of VariationAnalytical performanceultra-violetPhenylalanineLactoferrinlimit of quantificationlimit of detectionAccuracy profileIsoelectric pointhigh performance liquid chromatographyHPLC
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Sylvie Pochet, Céline Arnould, Perrine Debournoux, Jocelyne Flament, Odile Rolet-Répécaud, Eric Beuvier,