Detection of milk mixtures in Halloumi cheese.

A capillary electrophoresis method has been applied to the detection of illegal addition of milk from goat and/ or cow in Halloumi cheese, traditionally made with sheep milk. The electrophoretic profiles of the casein from Halloumi cheeses have revealed that caprine para-kappa-casein and bovine alphas1-casein peaks point to the presence of low percentages of goat's and/or cow's milk added to Halloumi cheese. Stepwise multiple linear regression has been used to predict these percentages with a standard error of the estimation of 2.14%. The analytical method combined with the statistical application is valid for the prediction of percentages higher than 2% of goat's and percentages of 5% of cow's milk added to the cheese either in fresh or ripened cheese. The standard error of estimation was higher for the prediction of cow's milk than for goat's milk.

Plasmin activity in pressurized milk.

The effects of pressure (up to 400 MPa), applied at room temperature, on native proteinase activity of milk were investigated by means of plasmin activity, plasmin-derived activity after plasminogen activation and their distribution in different milk fractions, micelle microstructure, beta-LG denaturation, and casein susceptibility to proteolytic attack. The pressure conditions assayed did not lead to plasmin inactivation and only decreased around 20 to 30% total plasmin activity after plasminogen activation. However, pressure caused severe disruption of the micellar structure, releasing high levels of caseins, plasmin, and plasminogen to the soluble fraction of milk. High levels of soluble denatured beta-LG were also found in the ultracentrifugation supernatants of pressurized milks, particularly in those treated at 400 MPa. Probably as a result of micellar disintegration, caseins became more susceptible to proteolysis by exogenous plasmin. However, no enhanced proteolytic degradation was observed when we compared the evolution of pressurized and unpressurized milks during refrigerated storage. Serum-liberated plasmin may become more vulnerable to the action of proteinase inhibitors leading to a reduced proteolysis on refrigerated storage, despite the increased susceptibility of caseins to proteinase action.

Micellar changes induced by high pressure. influence in the proteolytic activity and organoleptic properties of milk.

We investigated the effects of high-pressure treat-of 25 to 60 degrees C, on micelle structure, proteolytic activity, and sensory properties of milk. Pressure treatments at 25 degrees C considerably reduced micelle size, while pressurization at higher temperatures progressively increased micelle dimensions. Pressure-induced denaturation of beta-lactoglobulin (beta-LG) amounted 76% at 25 degrees C and was almost 100% in milks treated at 40 to 60 degrees C. alpha -Lactalbumin (alpha-LA) was resistant to pressure at temperatures up to 40 degrees C, but its denaturation reached 56% at 60 degrees C. Plasmin resisted pressurization at room temperature; however, pressure treatments at higher temperatures increased plasmin inactivation, which reached 86.5% at 60 degrees C. Pressurization at temperatures from 40 to 60 degrees C reduced the proteolytic activity and improved the organoleptical properties of milk, compared with the same treatments at 25 degrees C, which suggested that these combined treatments could be used to produce milk of good sensory properties with an increased shelf life. These results are discussed in the light of the changes found in micellar structure.

Detection of the presence of soya protein in milk powder by sodium dodecyl sulfate capillary electrophoresis.

Two different commercial kits for sodium dodecyl sulfate capillary electrophoresis (SDS-CE) were evaluated for the detection of the presence of soya protein in milk powder. The results obtained showed that SDS-CE allowed the separation of the basic subunits of glycinin and the alpha and alpha' subunits of beta-conglycinin from the main milk protein peaks. However, a detection limit lower than 10% (w/w) of soya protein in total protein could not be achieved. The use of a tetraborate-EDTA sample treatment minimized interferences from milk proteins, allowing the detection of at least 1% (w/w) of soya protein in total protein. The addition of soya protein hydrolysates could not be determined using SDS-CE.

Microbiological and chemical changes in high-pressure-treated milk during refrigerated storage.

The microbiological and biochemical changes during storage of high-pressure-treated (400 MPa at 25 degrees C, for 30 min) whole (3.5% fat) and skim (0.3% fat) milk at refrigeration temperatures (7 degrees C) were studied. From a microbiological point of view, high-pressure treatment of milk led to an increase in the shelf life because, after 45 days of refrigerated storage, the psychotrophic and pseudomonad counts of the pressurized milk were lower than those of the unpressurized milk after 15 days. Capillary electrophoresis of the case in fraction showed that proteolysis by bacterial proteinases was not relevant in high-pressure-treated milk, as evidenced by a negligible degradation of kappa-casein. However, since the pressure conditions assayed did not lead to plasmin inactivation, considerable beta-, alpha-s1-casein hydrolysis took place during refrigerated storage, which can be responsible for flavor defects. No significant differences were found between skim and whole high-pressure-treated-milks.