A rapid HPLC method for the determination of lactoferrin in milk of various species

Abstract

This Research Communication describes the adaptation and testing of an RP-HPLC method, previously tested for the determination of lactoferrin (LF) in whey, for its applicability to determine milk lactoferrin content. Milk samples of various species, namely, ovine, caprine, bovine, donkey and human milk, were tested. The advantage of this RP-HPLC method includes speed and convenience, as it does not include extensive pretreatment or separation steps. A simple pre-treatment step was added in order to remove fat and proteins of the casein family and the samples were tested. The results varied in terms of elution of the LF peak both between the milk of the different species as well as from the initial application on whey. The peak resolution was satisfactory in the cases of ovine, bovine and donkey milk samples while for caprine and human milk an interference with other peaks was observed. Nevertheless, quantification of LF was found possible for all samples. This new application of the modified method will allow the determination of LF in milk samples of the tested species either for everyday analysis or as a useful qualitative screening for presence or absence of LF.

Potential Protective Protein Components of Cow’s Milk against Certain Tumor Entities

“…In addition to identifying the possible health-promoting and anticarcinogenic properties of individual milk proteins, their quantification in milk of different species is also gaining importance. For example, goat milk contains far more Lf than cow milk, while the Lf content of human and sheep milk is also on a high level [139].…”

An Efficient Heparin Affinity Column Purification Method Coupled with Ultraperformance Liquid Chromatography for the Quantification of Native Lactoferrin in Breast Milk

“…Common quantification methods can be classified into immunological and nonimmunological methods [12]. Immunological methods include enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance-based immunosensors [13]. Although immunological methods exhibit high selectivity and sensitivity, they have poor reproducibility [14].…”

Bovine Lactoferrin Quantification in Dairy Products by a Simple Immunoaffinity Magnetic Purification Method Coupled with High-Performance Liquid Chromatography with Fluorescence Detection

“…23 Until now, the methodology for quantitation of bLf in dairy products has not been standardized. 9 The major challenges come from its low content compared to other abundant proteins and the complicated matrices of dairy products. Therefore, a highly specific and sensitive method for the extraction and determination of bLf is required at an intact level in dairy products.…”

“…Currently, bLf has been determined by separation techniques including capillary electrophoresis (CE), , high-performance liquid chromatography (HPLC), − and liquid chromatography-tandem mass spectrometry (LC–MS/MS). , Although CE has a broad use in protein analysis, its notable limitation is insufficient separation and unsatisfactory reproducibility. High-performance liquid chromatography coupled with an ultraviolet–visible detector (HPLC-UV) was employed to analyze bLf in milk and dairy products.…”

“…Magnetic bead-based immunoaffinity capture, as a simple and specific protein selection method, has increasingly been used in immunoassay protocols in recent years because it shows good coupling efficiency, operational stability, and reservation activity of the antibody. , Previous reports described that therapeutic protein could be selectively purified from the biological matrix by magnetic beads immobilized with the antibody. , Recently, Lf from human saliva was isolated using antibody-modified magnetic particles . Until now, the methodology for quantitation of bLf in dairy products has not been standardized . The major challenges come from its low content compared to other abundant proteins and the complicated matrices of dairy products.…”