title: Short communication: determination of lactoferrin in Feta cheese whey with reversed-phase high-performance liquid chromatography.
abstract: In the current paper, a method is introduced to determine lactoferrin in sweet whey using reversed-phase HPLC without any pretreatment of the samples or use of a separation technique. As a starting point, the most common HPLC protocols for acid whey, which included pretreatment of the whey along with a sodium dodecyl sulfate-PAGE step, were tested. By skipping the pretreatment and the separation steps while altering the gradient profile, different chromatographs were obtained that proved to be equally efficient to determine lactoferrin. For this novel 1-step reversed-phase HPLC method, repeatability was very high over a wide range of concentrations (1.88% intraday to 5.89% interday). The limit of detection was 35.46μg/mL [signal:noise ratio (S/N)=3], whereas the limit of quantification was 50.86μg/mL (S/N=10). Omitting the pretreatment step caused a degradation of the column's lifetime (to approximately 2,000 samples). As a result, the lactoferrin elution time changed, but neither the accuracy nor the separation ability of the method was significantly influenced. We observed that this degradation could be easily avoided or detained by centrifuging the samples to remove fat or by extensive cleaning of the column after every 5 samples.
authors.affiliation: Department of Scienze Agro-Ambientali e Territoriali, Università degli Studi di Bari Aldo Moro, Via G. Amendola, 165/A, 70126 Bari, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Tsakali
authors.affiliation: Department of Biosystems Engineering, Technological Educational Institute of Larissa, Larissa 41110, Greece.
authors.forename: K
authors.initials: K
authors.lastname: Petrotos
authors.affiliation: Department of Oenology and Beverage Technology, Technological Educational Institute of Athens, Ag. Spyridonos St., Egaleo, 122 10 Athens, Greece.
authors.forename: A
authors.initials: A
authors.lastname: Chatzilazarou
authors.affiliation: Department of Food Technology, Technological Educational Institute of Athens, Ag. Spyridonos St., Egaleo, 122 10, Athens, Greece.
authors.forename: K
authors.initials: K
authors.lastname: Stamatopoulos
authors.affiliation: Department of Scienze Agro-Ambientali e Territoriali, Università degli Studi di Bari Aldo Moro, Via G. Amendola, 165/A, 70126 Bari, Italy.
authors.forename: A G
authors.initials: AG
authors.lastname: D'Alessandro
authors.affiliation: Department of Animal Production, Technological Educational Institute of Larissa, Larissa 41110, Greece.
authors.forename: P
authors.initials: P
authors.lastname: Goulas
authors.affiliation: Department of Food Science and Technology, Agricultural University of Athens, 75 Iera Odos St, Votanikos, 118 55, Athens, Greece.
authors.forename: T
authors.initials: T
authors.lastname: Massouras
authors.affiliation: Chemical Engineering Department, Katholieke Universiteit Leuven, W. de Croylaan 46, PB 2423, 3001 Leuven, Belgium. Electronic address: jan.vanimpe@cit.kuleuven.be.
authors.forename: J F M
authors.initials: JF
authors.lastname: Van Impe
mesh_terms: D002611:Cheese / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004591:Electrophoresis, Polyacrylamide Gel; D005504:Food Analysis / Q000379:methods*; D007781:Lactoferrin / Q000032:analysis*
chemical_list: D007781:Lactoferrin
keywords: Feta cheese; lactoferrin; reversed-phase HPLC; whey