Quantitative analysis of major whey proteins by capillary electrophoresis using uncoated capillaries.
A method that allows separation and quantitation of the main whey proteins by capillary electrophoresis using uncoated capillaries is proposed. Separations are performed using 100 mM borate buffer, pH 8.2, containing 30 mM sodium sulfate. The use of high pH and high ionic strength buffer reduces adsorption of proteins on the capillary wall, making their separation possible. Reproducibility of migration times and areas of peaks are improved by optimizing the capillary equilibration protocol and by using an internal standard. Relative standard deviations ranging between 0.74 and 1.03% for migration times and 2.14 to 5.23% for areas of major peaks are obtained. Detection limits equal to or lower than 0.5 mg/100 mL are achieved. Linear relationships of peak area versus concentration have been used to quantitate bovine serum albumin (BSA), alpha-LA (alpha-lactalbumin), beta-LG A (beta-lactoglobulin A) and beta-LG B (beta-lactoglobulin B) in cow's milk subjected to different thermal treatments. Electrophoretic profiles of these milk samples show peaks from other peptides besides those from main proteins. Characteristic patterns for whey from different species are obtained.
Recio
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Instituto de Fermentaciones Industriales CSIC, Madrid, Spain.
Molina
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Ramos
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de Frutos
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D000818:Animals; D002417:Cattle; D004586:Electrophoresis / Q000379:methods*; D006041:Goats; D006801:Humans; D007768:Lactalbumin / Q000032:analysis; D007782:Lactoglobulins / Q000032:analysis; D008894:Milk Proteins / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000032:analysis; D012756:Sheep; D012822:Silicon Dioxide; D000067816:Whey Proteins
D007782:Lactoglobulins; D008894:Milk Proteins; D000067816:Whey Proteins; D012710:Serum Albumin, Bovine; D012822:Silicon Dioxide; D007768:Lactalbumin