Lactoferrin in breast milk-based powders.

This study aimed to determine lactoferrin (LF) in breast milk-based powders and formulas. Lactoferrin is an important whey protein in all mammalian milks and is responsible in large part for the known antimicrobial effects of human milk in particular. As breast feeding is not always possible, formulas based on cows milk have been developed in order to meet the nutritional needs of the newborn, while more recently human breast milk-based powders have been introduced to offer the biological functionality of human milk to pre-term and critically ill babies. In the present work, the amount of LF in commercial breast milk-based powders was tested by a validated RF-HPLC method for the determination of LF in breast milk in order to examine both the applicability of the method but at a second level the amount of LF in these commercial products. The detection of LF was possible but the complexity of the matrix lead us to the use the standard addition methodology in order to achieve quantification. The results indicated that breast milk-based powders had higher amount of LF than cows milk-based formulas, both non-fortified and fortified.

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

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.

Evaluation of the suitability of low hazard surfactants for the separation of phenols and carotenoids from red-flesh orange juice and olive mill wastewater using cloud point extraction.

Natural antioxidants derived from plant sources attract considerable scientific interest. While classic extraction methods consume high volumes of toxic organic solvents, cloud point extraction requires surfactant not exceeding 15% of the waste volume. In preliminary tests, the suitability of various low hazard surfactants (Span 20, PEG 400, Tween 80 and 20) was explored for separation of phenols and carotenoids from olive mill wastewater and red-flesh orange juice. Tween 80 showed the highest recovery and further applied to the next experiments. The most appropriate surfactant concentrations were 5% (for olive mill wastewater) and 7% (for orange juice) as indicated by recovery % and the rest cloud point extraction parameters (analyte concentration, concentration factor, and phase volume ratio). A double step CPE with 5% + 5% of Tween 80 recovered up to 94.4% of the total phenols from olive mill wastewater, while a 7% + 7% of Tween 80 recovered up to 72.4% of the total carotenoids from orange juice. Evaluation of the final effects and extraction efficiency of single and double step cloud point extraction shows that double step scheme seems to be preferable in both cases. Finally, phenols and carotenoids recovered by Tween 80 maintained high antiradical activity (DPPH test).

Impact of tannin addition on the antioxidant activity and sensory character of Malagousia white wine.

Enological tannins are assessed as promising alternative to SO2 in order to control oxidative process during winemaking, due to allergic reactions incurred by sulfite sensitive individuals. In the present study, the commercial enological Tara tannin "Vitanil B″ was added, as alternative to the addition of sulfites, at different concentrations (100-500 mg/L) in white wine from grapes of Vitis vinifera L. var. Malagousia in order to enhance antioxidant stability and sensory character of the wine. Considering photometric analyses and chromatic parameters results, tannin addition (300 mg/L) in Malagousia enhanced total phenolic content, antioxidant and antiradical activity and prevented color deterioration, for a storage period of 100 d, compared to control and sulfited wines. Moreover, aroma quality, body, after taste and overall acceptance of wine treated with 300 mg/L tannin, were highly appreciated and received the highest scores. The overall evaluation of tannin addition was performed by Principal Component Analysis, leading to discrimination of wines, according to photometric, color and sensory analysis parameters. Conclusively, tannin addition resulted in a considerable increase of total phenolic content, antioxidant and antiradical activity, compared to the control and sulfited wines, maintaining the sensory parameters and overall acceptance of Malagousia wine.

Removal of polyphenols from wine sludge using cloud point extraction.

Cloud point extraction (CPE), a promising and simple technique for the separation of organic compounds using surfactants (Genapol X-080 [oligoethylene glycol monoalkyl ether] and PEG 8000 [polyethylene glycol with molecular weight of 8000]), was used to recover polyphenols from wine sludge (wine production waste). The effect of various parameters such as surfactant concentration, temperature, and pH on the percentage of phenol recovery and phase volume ratio during phenol separation from wine sludge was investigated, and the derived optimum parameters were used as the basis for the selection of CPE conditions. When a two-step CPE with a total of 4% v/v of Genapol X-080 (pH = 3.5, temperature = 55 degrees C, and time = 30 min) or 10% v/v of PEG 8000 (pH = 2.5, temperature = 55 degrees C, and time = 30 min) was applied the phenol recovery values achieved were 75.8 or 98.5%, respectively. Phenols recovered from wine sludge using the above surfactants maintained high antiradical activity as determined by the 1,1-diphenyl-2-picrylhydrazyl method.

Antioxidant Activity and Thermal Stability of Oleuropein and Related Phenolic Compounds of Olive Leaf Extract after Separation and Concentration by Salting-Out-Assisted Cloud Point Extraction.

A fast, clean, energy-saving, non-toxic method for the stabilization of the antioxidant activity and the improvement of the thermal stability of oleuropein and related phenolic compounds separated from olive leaf extract via salting-out-assisted cloud point extraction (CPE) was developed using Tween 80. The process was based on the decrease of the solubility of polyphenols and the lowering of the cloud point temperature of Tween 80 due to the presence of elevated amounts of sulfates (salting-out) and the separation from the bulk solution with centrifugation. The optimum conditions were chosen based on polyphenols recovery (%), phase volume ratio (Vs/Vw) and concentration factor (Fc). The maximum recovery of polyphenols was in total 95.9%; Vs/Vw was 0.075 and Fc was 15 at the following conditions: pH 2.6, ambient temperature (25 °C), 4% Tween 80 (w/v), 35% Na2SO4 (w/v) and a settling time of 5 min. The total recovery of oleuropein, hydroxytyrosol, luteolin-7-O-glucoside, verbascoside and apigenin-7-O-glucoside, at optimum conditions, was 99.8%, 93.0%, 87.6%, 99.3% and 100.0%, respectively. Polyphenolic compounds entrapped in the surfactant-rich phase (Vs) showed higher thermal stability (activation energy (Ea) 23.8 kJ/mol) compared to non-entrapped ones (Ea 76.5 kJ/mol). The antioxidant activity of separated polyphenols remained unaffected as determined by the 1,1-diphenyl-2-picrylhydrazyl method.

Improvement of oleuropein extractability by optimising steam blanching process as pre-treatment of olive leaf extraction via response surface methodology.

Impact of steam, hot water blanching and UV-C irradiation as pre-treatments on extraction of oleuropein and related biophenols from olive leaves (OLs), was investigated. Moreover, particle size effect of olive leaves and steam blanching duration were selected as independent variables to optimise steam blanching process in terms of oleuropein content (OC) and antioxidant activity (AC) of ethanolic extracts, by using response surface methodology. Optimum conditions for OC and AC were 10 min steam blanching of 20-11 and 3-1mm olive leaf fraction, respectively. Depending on the extraction procedure, at optimum conditions of steaming the results indicate that steam blanching of OL prior to extraction can significantly increase oleuropein yield from 25 to 35 times compared to non-steam blanched sample, whereas the antioxidant activity increased from 4 to 13 times. No significant UV-C effect was observed in OC and AC, while hot water blanched samples showed significantly higher oleuropein yields and antioxidant activity compared to untreated samples.