Ferrous bisglycinate content and release in W1/O/W2 multiple emulsions stabilized by protein–polysaccharide complexes

Ferrous bisglycinate aqueous solution was entrapped in the inner phase (W₁) of water-in-oil-in-water (W₁/O/W₂) multiple emulsions. The primary ferrous bisglycinate aqueous solution-in-mineral oil (W₁/O) emulsion contained 15% (w/w) ferrous bisglycinate, had a dispersed phase mass fraction of 0.5, and was stabilized with a mixture of Grindsted PGPR 90:Panodan SDK (6:4 ratio) with a total emulsifiers concentration of 5% (w/w). This primary emulsion was re-emulsified in order to prepare W₁/O/W₂ multiple emulsions, with a dispersed mass fraction of 0.2, and stabilized using protein (whey protein concentrate (WPC)):polysaccharide (gum arabic (GA) or mesquite gum (MG) or low methoxyl pectin (LMP)) complexes (2:1 ratio) in the W₂ aqueous phase. The W₁/O/W₂ multiple emulsion stabilized with WPC:MG (5% w/w total biopolymers concentration) provided smaller droplet sizes (2.05 μm), lower rate of droplet coalescence (7.09 × 10⁻⁷ s⁻¹), better protection against ferrous bisglycinate oxidation (29.75% Fe³⁺) and slower rate of ferrous bisglycinate release from W₁ to W₂ (K_H = 0.69 mg mL⁻¹ min^−0.5 in the first 24 h and 0.07 mg mL⁻¹ min^−0.5 for the next 19 days of storage time). Better encapsulation efficiencies, enhanced protection against oxidation and slower release rates of ferrous bisglycinate were achieved as the molecular weight of the polysaccharide making up protein:polysaccharide complex was higher. Thus, the factor that probably affected most the overall functionality of multiple emulsions was the thickness of the complex adsorbed around the multiple emulsion oil droplets. These thicknesses determined indirectly by measuring the z-average diameter of the complexes, and that of the WPC:MG (529.4 nm) was the largest.     

Reduced-fat white fresh cheese-like products obtained from W1/O/W2 multiple emulsions: Viscoelastic and high-resolution image analyses

This paper focuses on the study of the rheological and structural changes of reduced fat cheeses (EC) obtained by substituting milk-fat by multiple emulsions (W₁/O/W₂) stabilized with different hydrocolloids in comparison to a full-fat white fresh cheese (WFC). Amidated low-methoxyl pectin, carboxymethylcellulose (CMC), gum Arabic (GA), and blends of these hydrocolloids were used as emulsion stabilizing agents. Cheeses storage (G′) and loss (G″) moduli as a function of strain % were determined and the ratio of tan δ in the non-lineal viscoelastic region (tan δ_B) to that in the lineal viscoelastic region (tan δ_A) was obtained. The EC cheeses made from W₁/O/W₂ emulsions with GA (EC_GA) and CMC (EC_CMC) exhibited non-significantly different tan δ_B/tan δ_A ratio values from that of WFC cheese. Scanning electron micrographs (SEM) showed that all the cheeses exhibited different surface microstructure. Detrended fluctuation analysis (DFA) of SEM micrographs indicated that all cheeses structure displayed a hierarchical configuration, from well-ordered particles in the small scale range to randomly dispersed particles in the high scale range. The EC_GA and EC_CMC cheeses also showed similar particle distribution over the particle size range of 20–60 μm to that of the WFC cheese.     

Effect of Maillard reaction products derived from the hydrolysate of mechanically deboned chicken residue on the antioxidant,textural and sensory properties of Cantonese sausages

Protein hydrolysates as precursors of Maillard reaction were obtained via enzymatic hydrolysis of mechanically deboned chicken residue (MDCR). The Maillard reaction products (MRPs) were prepared at 90 (M1), 100 (M2), 110 (M3) and 120 °C (M4), respectively. MRPs possessed a strong reducing power and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. According to the evolution of total free fatty acids and peroxide value of Cantonese sausage with MRPs during storage, M1 and M3 had a potent antioxidant activity (P < 0.05) due to their antioxidant abilities and inhibitory action against lipolytic enzymes. Cantonese sausages treated with M1 and M2 showed good textural and sensory properties. However, M3 and M4 had a negative (P < 0.05) effect on the flavour and texture of Cantonese sausages compared to control. The results suggested that M1 was very potential to be used to improve their antioxidant, textural and sensory quality.     

The effect of storage on nutritional,textural and sensory characteristics of creamy ricotta made from whey as well as cow's milk and goat's milk

The aim of this study was to develop a creamy ricotta using a mixture of goat and cow whey as the main ingredients, with the addition of whole goat and cow milk. The nutritional composition, texture and sensory characteristics of the ricotta cheese were evaluated over 14 days of refrigerated storage. There was a decrease in protein and ash content and pH changes during the assessed storage periods. The instrumental texture profile indicated that the creamy ricotta was easily deformable, with minimal inelasticity and a cohesive, soft and delicate texture. The medium‐ and long‐chain fatty acid content was higher than the short‐chain fatty acid content. The flavour and aroma of creamy ricotta were described by the panel as having a soft texture and wet appearance. The use of whey from goat and cow cheese in making creamy ricotta is feasible and does not cause negative impact in the final product.     

Double emulsions (W/O/W): physical characteristics and perceived intensity of salty taste

This study addressed the correlation between physical characteristics of double emulsions and sensory perception, as the microstructure of these systems may provide the mechanism to understanding the initial cause of the altered salty taste. Thus, double emulsions (W/O/W) were prepared using different volumes of the internal aqueous phase while maintaining the same fat and sodium contents in the evaluated systems. Polyglycerol polyricinoleate (PGPR) and tween 80 were used as hydrophobic and hydrophilic emulsifiers, respectively. After preparation, the samples were stored at 25 °C for 4 days and submitted to analysis of optical microscopy, distribution and polydispersity of the oil droplets size, electrical conductivity, rheological behaviour and sensorial analysis. It was found that the use of different emulsifier concentrations (PGPR) did not influence the physical characteristics of the emulsions with the same formulation, but emulsions with different internal aqueous phase concentrations presented different results. These distinct characteristics may have influenced sensorial perception, as the emulsion with higher internal phase concentration was considered saltier. Thus, it can be concluded that structural differences of the double emulsions can be used to decrease the sodium contents without perceivable changes in salty taste.     

免疫亲和层析净化-高效液相色谱法同时测定牛奶和奶粉中黄曲霉毒素B1、B2、G1、G2、M1、M2

建立牛奶和奶粉中黄曲霉毒素B1、B2、G1、G2、M1、M2的免疫亲和层析净化和柱后衍生高效液相色谱测定方法。样品经溶解、离心、过滤后,通过免疫亲和柱,黄曲霉毒素特异性抗体选择性地与存在的黄曲霉毒素抗原键合,形成抗体-抗原复合体。甲醇-乙腈混合溶液(4:5,v:v)洗脱,带荧光检测器的高效液相色谱仪经柱后衍生测定,外标法定量。标准曲线线性良好,添加回收率在57.0%～88.7%,相对标准偏差在3.37%～16.9%,牛奶中各黄曲霉毒素检出限:B1为2ng/kg,B2为1ng/kg,G1、G2为3ng/kg,M1、M2为5ng/kg;奶粉中B1为20ng/kg,B2为10ng/kg,G1、G2为30ng/kg,M1、M2为50ng/kg,检测低限能够满足各国对牛奶和奶粉中黄曲霉毒素的限量要求。该方法准确、快速、灵敏度高,适用于牛奶和奶粉中黄曲霉毒素的测定。     

Changes in textural, microstructural, and colour characteristics during ripening of cheeses made from raw, pasteurized or high-pressure-treated goats’ milk

Goats’ milk cheeses were made from raw (RA), pasteurized (PA; 72°C, 15 s) or pressure-treated (PR; 500 MPa, 15 min, 20°C) milk to compare textural, microstructural, and colour characteristics in relation to ripening time. Texture, microstructure and colour were evaluated by uniaxial compression and stress relaxation tests, confocal laser scanning microscopy and Hunter colorimetry, respectively.Raw and PR cheeses were firmer and less fracturable than PA cheese, but differences became less notable toward the end of ripening. PA and PR cheeses were less cohesive than RA cheese. Although cheeses exhibited a loss of elastic characteristics with ageing, PR cheese showed the most elastic behaviour initially. Confocal laser scanning micrographs displayed PR cheese with a regular and compact protein matrix, with small and uniform fat globules resembling the structure of RA cheese. Finally, colour evaluation demonstrated significant differences between cheeses due to milk treatments and ripening time.     

Technologically important characteristics of Enterococcus isolates from milk and dairy products

Enterococcus faecium (54 strains), E. faecalis (40 strains), and E. durans (14) were isolated from various dairy products (raw milk, cream, butter and fermented milk products) during a previous study (Wessels et al., 1988). In this article various characteristics of these isolates, which may have a bearing on their significance in dairy products, have been studied. A large percentage of the identified strains of all three species were able to grow at 7°C. Seventy-six percent of the E. faecium strains, 62% E. faecalis and 50% E. durans strains also showed proteolytic activity at psychrotrophic temperatures. The fact that proteolytic activity could be detected within 2 days at 7°C is significant, since bulk cooled milk is normally held for 3 to 4 days at temperatures between 4 and 7°C at farms or factories prior to processing. This examination confirmed that enterococci are proteolytic rather than lipolytic.     

An ultra-sensitive monoclonal antibody-based competitive enzyme immunoassay for aflatoxin M1 in milk and infant milk products

A sensitive and specific monoclonal antibody (Mab) against aflatoxin M₁ (AFM₁), named as 2C9, was selected by semi-solid HAT medium. It exhibited high affinity for AFM₁ of 1.74 × 10⁹ L/mol and no cross-reactivity to aflatoxin B₁, B₂, G₁ and G₂. Based on the antibody, an ultra-sensitive competitive enzyme-linked immunosorbent assay (ELISA) was developed for AFM₁ in milk and infant milk products. Assays were performed in the AFM₁-BSA coated (0.0625 μg/mL) ELISA format in which the antibody was diluted 1:10,000. Several physicochemical factors (pH, ionic strength and blocking solution) that influence assay performance were optimised. Finally, the limits of detection were 3 ng/L for milk and 6 ng/L for milk-based cereal weaning food, inter-assay and intra-assay variations were less than 10%, and the recovery ranged from 91% to 110%. Thirty samples were analysed, and concordant results were obtained when the data were compared with a reference high-performance liquid chromatography method.     

免疫初乳中IgG1和IgG2的分离纯化

采用硫酸胺盐析,DE－52离子交换色谱,SephacryIS-100凝胶过滤色谱从免疫初乳分离纯化出了IgG1和IgG2,经SDS－PAGE电泳及免疫电泳证实为纯品,并对IgG1和IgG2的抗体凝集价进行了测定。     

Evaluation and Improvement of Extraction Methods for the Analysis of Aflatoxins B1, B2, G1 and G2 from Naturally Contaminated Maize

The extraction procedure for aflatoxin determination in maize is based on a methanol–water (8 + 2 v/v) or an acetone–water (85 + 15 v/v) mixture. Initially, the extraction efficiency of two solvents was evaluated for each aflatoxin. Different results were obtained for highly contaminated maize: significantly higher levels of aflatoxin B₁ were obtained by acetone–water, on the contrary higher levels of aflatoxin G₂ were achieved by methanol–water. Then, acetone–water mixtures in different proportions (7 + 3, 6 + 4 and 5 + 5 v/v) were tested to improve the extraction of aflatoxin G₂. Applying these extraction mixtures, the values both of aflatoxin B₁ and of other aflatoxins were generally higher compared to those obtained by acetone–water 85 + 15; moreover, acetone–water (6 + 4) and (7 + 3) showed the best extraction efficiency for all aflatoxins.     

Dynamic supercritical CO2 extraction for removal of cholesterol from anhydrous milk fat

The feasibility of different extraction techniques for the selective removal of cholesterol from anhydrous milk fat (AMF) by supercritical carbon dioxide has been studied. A dynamic extraction system was used to determine both the experimental solubility of anhydrous milk fat in the supercritical solvent and the selectivity of cholesterol over anhydrous milk fat at 40–70°C and 8–40MPa at various stages during extraction. In addition, adsorbents were used for the selective removal of cholesterol from anhydrous milk fat. The results indicate that a direct extraction alone or with several separators in series are not practical, but a selective removal of 97% of the cholesterol from the extracted anhydrous milk fat is possible by using an adsorbent with recovery of solvent and cholesterol. A schematic industrial countercurrent process for the removal of cholesterol and the fractionation of milk fat is proposed.     

Distribution of aflatoxin M1 during Grana Padano cheese production from naturally contaminated milk

The distribution of aflatoxin M₁ (AFM₁) has been investigated in samples of whey, curd and a typical hard and long maturing cheese like Grana Padano (ripened for twelve months), produced with naturally contaminated milk in a range of 30–98 ng AFM₁/kg. AFM₁ determinations were carried out on 25 samples of each product by reverse-phase HPLC and fluorescence detection with post-column derivatisation, after a preliminary C18-SPE clean-up. Experimental results show that, in comparison to milk, AFM₁ concentration levels increased both in curd (3-fold) and in long maturing cheese (4.5-fold), while AFM₁ occurrence in whey decreased by 40%.     

Production of enniatins A,A1, B,B1, B4, J1 by Fusarium tricinctum in solid corn culture: Structural analysis and effects on mitochondrial respiration

Enniatins (ENs) are secondary fungal metabolites with hexadepsipeptidic chemical structure and they possess a number of potent biological activities that can contaminate several kind of food and foodstuffs increasing the exposure risk for consumers. ENs are produced by several Fusariun strains including Fusarium subglutinans, Fusarium proliferatum and Fusarium tricinctum. Production of a mixture of ENs was performed by culturing F. tricinctum ITEM 9496 on white corn as substrate. The solid culture components were dried and subsequently extracted with water/methanol (50/50 v/v, 0.5% NaCl), homogenised, filtered, extracted by ethyl acetate and analysed by liquid chromatography with diode array detection (LC-DAD). The crude extract was first separated by low pressure liquid chromatography (LPLC) and then further purified by liquid chromatography (LC), resulting in six compounds with a purity higher than 95% as calculated by ¹H NMR, and with a yield of 30–300 mg per compound. The chemical structures of the ENs were determined by liquid chromatography coupled to mass spectrometry (LC–MS) and nuclear magnetic resonance (NMR). The biological activity of the resulting ENs was determined using a mitochondrial respiration test. We discovered that all the ENs studied induced an increase in the mitochondrial respiration resulting in uncoupling of the oxidative phosphorylation. This effect was most likely due to flux of K⁺ ions into the mitochondrial matrix. The order of potency of the ENs derivatives was: A₁ > B₁ > B > A > B₄ > J₁. These results suggest a correlation between the chemical structures and bioactivity and confirm the severe risks for human associated with consumption of enniatins.     

快速测定婴幼儿配方乳粉中的维生素B1及B2

建立高效液相色谱-荧光法快速测定婴幼儿配方乳粉中维生素B₁及维生素B₂。方法优化了前处理条件,采用高压酸提法提取目标化合物,样品沉淀蛋白后直接测定维生素B₂,取部分清液用碱性铁氰化钾衍生后测定维生素B₁。目标物在对应的浓度范围内呈现良好的线性关系（R²≥0.9998）,样品加标回收率在92.4%～102%之间。该方法快速,准确,适用于婴幼儿配方乳粉中维生素B₁、B₂的测定。     

Chemiluminescent imaging detection of staphylococcal enterotoxin C1 in milk and water samples

A sensitive, simple and rapid technique for high throughput simultaneous detection of staphylococcal enterotoxin C₁ (SEC₁) has been developed. The proposed method has the advantage of showing the specificity of enzyme-linked immunosorbent assays (ELISA), sensitivity of luminol-based enhanced chemiluminescence (ECL) assay, and high throughput of chemiluminescence (CL) imaging assay. It was based on a standard sandwich immunoassay format; 96-well ELISA plates were used as solid phase material. A commercial high-sensitivity cooled CCD camera has been applied to image the weak CL. Under the optimum conditions, the increased CL intensity was proportional with the concentration of SEC₁ in the range of 8.0–125.0 ng ml⁻¹ and the detection limit was 0.5 ng ml⁻¹ (3σ). The relative standard deviation (RSD) for eight parallel measurements of 25.0 ng ml⁻¹ SEC₁ was 0.06. The proposed method has been successfully applied to the determination of SEC₁ in milk and water samples. The results obtained compared well with those by ELISA.     

Evaluation of some plant products against Trogoderma granarium everts in sorghum and their effects on nutritional composition and organoleptic characteristics

The efficacy of neem, citrus, garlic and podina powders and neem oil at 1 and 2% level (w/w) in sorghum grains was evaluated against larvae of Trogoderma granarium. Neem kernel powder and neem oil completely prevented damage by an introduced larval population. In other treatments, grain damage was noticeable after 3 months, although significantly less than in untreated grains. Thereafter, the percentage of damaged kernels increased progressively and reached as high as 29% (neem leaf), 44% (citrus), 56% (podina) and 71% (garlic) compared to 85% in the control after 6 months. Nutritional composition viz., moisture, ash, fibre, fat, protein and carbohydrates of treated grains remained unaffected after 1 month of storage. After 6 months, changes in nutritional composition were proportional to insect damage. Chapatis (unleavened bread) prepared from flour of treated grains after 1 and 6 months were of normal colour, appearance and texture, but in all cases, their taste, aroma and overall acceptability were adversely affected either by insect infestation or plant products or their combination.     

Effects of reverse CO2 acidification cycles,calcium supplementation,pH adjustment and chilled storage on physico-chemical and rennet coagulation properties of reconstituted low- and medium-heat skim milk powders

The reversibility extent of one and two reverse CO₂ acidification cycles on the physico-chemical and rennet coagulation properties of milks reconstituted from low- (LH) or medium- (MH) heat skim powder, enriched or not with calcium and pH adjusted or not was investigated. The ionized calcium concentration, buffering properties and average casein micelle size of untreated and CO₂-treated milks were evaluated before and after a chilled storage for 2 days. The ionized calcium concentration and buffering properties have been modified by the CO₂-treatment, particularly after a second CO₂-cycle. These modifications were highly dependent on the initial milk properties and chilled storage. Inversely, the average casein micelle size was not significantly changed. In addition, the rennet-clotting behaviour checked by near infrared spectroscopy (NIR-S) and rheology (SAOR) indicated the main factors responsible for changes in the casein micelles environment and dynamic casein micellar calcium phosphate reorganization, especially after two CO₂-cycles. A single CO₂-cycle induced a better rennetability for non Ca-enriched milk reconstituted from MH-powder. A second CO₂-cycle was particularly efficient to improve Ca-enriched pH-adjusted milks.     

Effects of cysteine and different incubation temperatures on the microflora,chemical composition and sensory characteristics of bio-yogurt made from goat’s milk

Yogurt and bio-yogurts were made from goat’s milk using a starter culture and probiotic culture, with or without cysteine addition (0.5%). Incubation was carried out at 37 and 42 °C until pH 4.6 was reached and yogurts were stored 4 ± 1 °C for 14 days. Yogurts were analysed 1, 7 and 14 days after production. The addition of cysteine, incubation temperature and storage time significantly influenced overall properties of the samples. During storage, whey separation, pH and acetaldehyde decreased, while lactic acid increased. Viable bacterial counts in all bio-yogurts were above 10⁷ cfu g⁻¹ at the end of storage. Whey separation, titratable acidity and lactic acid contents were lower, while acetaldehyde and viable bacterial counts were higher in the bio-yogurts incubated at 37 °C and supplemented with cysteine, in comparison to other samples. This indicates that lower temperature incubation and addition of cysteine can be used satisfactorily for the manufacture of bio-yogurt.