Quenching Mechanisms and Kinetics of α-Tocopherol and β-Carotene on the Photosensitizing Effect of Synthetic Food Colorant FD&C Red No. 3

ABSTRACT: Effects of FD&C Red No. 40, Red No. 3, Yellow No. 5, Yellow No. 6, Green No. 3, Blue No. 1 and Blue No. 2 on 0.03M soybean oil oxidation in acetone at 25 °C under light were studied by measuring headspace oxygen depletion. As Red No. 3 increased from 0 to 5, 20, 100 and 200 ppm, the headspace oxygen was reduced by 2 to 70, 73, 77 and 77%, respectively, for 4 h. Only Red No. 3 acted as a photosensitizer to produce singlet oxygen in the oil. The quenching rates of α-tocopherol and β-carotene for the singlet oxygen by Red No.3 were 4.1 × 10⁷ M⁻¹s⁻¹ and 7.3 × 10⁹ M⁻¹s⁻¹, respectively. When β-carotene was below 1.86 × 10⁻⁶ M, β-carotene quenched singlet oxygen, but it quenched both singlet oxygen and Red No. 3 at or above 3.72 × 10⁻⁶ M. However, α-tocopherol quenched singlet oxygen only.     

Stability of α-, γ-, or δ-Tocopherol during Soybean Oil Oxidation

ABSTRACT:  The decomposition of α-, γ-, or δ-tocopherol in soybean oil during 24 d of storage at 50 °C was determined by high performance liquid chromatography (HPLC). The initial contents of α-, γ-, and δ-tocopherol in soybean oil were 53, 750, and 268 ppm, respectively. The degradation rates of α-, γ-, and δ-tocopherol for the first 10 d were 5.6%, 1.2, and 0.5% per day, respectively. The α-tocopherol was completely destroyed in 16 d. The destructions of γ- and δ-tocopherol were 28% and 17% after 24 d. The induction period of soybean oil determined by headspace oxygen, conjugated diene, and peroxide value was 8 d. As the degradations of α-, γ-, and δ-tocopherol increased, the headspace oxygen disappearance, conjugated diene formation, and peroxide value of soybean oil increased. The correlation coefficient between the degradation of tocopherols and the oxidation of soybean oil was about 0.95. The degradation of tocopherols in soybean oil during storage was due to the oxidation.     

Riboflavin Photosensitized Singlet Oxygen Oxidation of Vitamin D

Samples containing various levels of vitamin D and riboflavin, with and without ascorbic acid or a-tocopherol were prepared in a model system. Samples were stored in the light or in the dark at 45®C for up to 8 h. Headspace oxygen was determined by gas chromatography with thermal conductivity detection. Oxidation of vitamin D was not observed in samples without riboflavin stored in the light nor in samples with riboflavin stored in the dark. Vitamin D with riboflavin was oxidized under light. α-Tocopherol quenched singlet oxygen at a rate of 2.52 × 10⁸ M^-1 sec^-1, whereas ascorbic acid quenched singlet oxygen at a rate of 2.23 × 10⁷ M^-1 sec^-1.     

EFFECT OF SEVERAL EXPERIMENTAL PARAMETERS ON COMBINATION OF RED KIDNEY BEAN (PHASEOLUS VULGARIS) α-AMYLASE INHIBITOR WITH PORCINE PANCREATIC α-AMYLASE

Purified red kidney bean (Phaseolus vulgaris) amylase inhibitor forms a 1:1 stoichiometric complex with porcine pancreatic α-amylase leading to complete loss of enzyme activity on starch. Rate of complex formation is pH dependent and is maximal at pH 5. The rate constants for complex formation, as measured by loss of amylase activity, were 2.85 × 10⁴ M^-1 sec^-1 at pH 6.9 (ionic strength of 0.918) and 2.55 × 10⁵ M^-1 sec^-1 at pH 5 at 30°C. At pH 6.9, rate of complex formation was 4.8 times faster at 0.918 ionic strength as compared with the rate at 0.138 ionic strength. At 30°C, pH 6.9 and ionic strength of 0.168 the dissociation constant of the enzyme-inhibitor complex was determined to be 3.5 × 10^-11 M. The rate constant for dissociation of the complex was calculated to be 8.7 × 10^-8 sec^-1 under the same conditions. The rate constant for complex formation, at ionic strength of 0.168, was 1.1 × 10⁴ M^-1 sec^-1 at 37⁰ and 9.77 × 10² M^-1 sec^-1 at 25.7°C. The calculated activation energy for complex formation is 39.5 kcal/mole suggesting a rate-controlling conformational change. Oxidation of the carbohydrate moiety of the glycoprotein inhibitor caused complete loss of activity. Maltose, a competitive inhibitor of α-amylase, bound as readily to the enzyme-inhibitor complex as to free α-amylase. Trypsinized α-amylase, although still able to bind to Sephadex, did not bind inhibitor. The experiments with maltose and trypsinized amylase suggest the inhibitor may not bind at the active site of α-amylase.     

Kinetics for Singlet Oxygen Formation by Riboflavin Photosensitization and the Reaction between Riboflavin and Singlet Oxygen

ABSTRACT: The formation of singlet oxygen by riboflavin and the kinetics and mechanisms of riboflavin degradation in aqueous solution under light were determined. The singlet oxygen formation rate by riboflavin was 2.31 μmole oxygen/mL headspace/h of serum bottle. The degradations of riboflavin were 66% in D₂O and 40% in H₂O, respectively, under light after 24 h. The results indicate that singlet oxygen is involved in riboflavin destruction under light. The riboflavin destructions were 94.0% and 15.7% with 0 mM or 160 mM ascorbic acid, respectively, under light after 96 h. The reaction rate between riboflavin and singlet oxygen was 1.01 × 10¹⁰/M/s, which is a diffusion-controlled reaction rate. This explains the extremely fast degradation of riboflavin in foods under light. Ascorbic acid and sodium azide reduce the degradation of riboflavin under light with different quenching mechanisms. Ascorbic acid quenched both singlet oxygen and excited triplet riboflavin. Sodium azide quenched only the singlet oxygen in riboflavin solution with a quenching rate of 1.547 × 10⁷/M/s. With the involvement of both the Type-I and Type-II mechanisms in the riboflavin degradation under light, singlet oxygen quencher alone could not protect the riboflavin from degradation completely. Addition of ascorbic acid can protect riboflavin oxidation in foods exposed to light.     

Quenching Mechanism and Kinetics of Ascorbic Acid on the Photosensitizing Effects of Synthetic Food Colorant FD&C Red Nr 3

ABSTRACT:  The pH effect on the oxidative stability of ascorbic acid in the presence of food colorant FD&C Red Nr 3 during storage with or without light was investigated. The quenching mechanism and kinetics of ascorbic acid on the FD&C Red Nr 3 photosensitized oxidation in an aqueous system at 25 °C were also studied by measuring the degradation of ascorbic acid or depletion of headspace oxygen. Red Nr 3 had no influence on the oxidation of ascorbic acid under dark storage, but accelerated its oxidation rate under light storage. The oxidative stability of ascorbic acid decreased as the pH increased from 4 to 7 under light without FD&C Red Nr 3. The quenching rates of ascorbic acid on the singlet oxygen by measuring the degradation of ascorbic acid in the presence of Red Nr 3 under light storage were 1.53 ± 0.15 × 10⁸, 1.86 ± 0.25 × 10⁸, and 1.19 ± 0.12 × 10⁸ M⁻¹S⁻¹ at pH 4, 5.6, and 7, respectively.     

Effect of pan-frying in extra-virgin olive oil on odour profile, volatile compounds and vitamins

Changes in odour of Arauco (ARA) and Arbequina (ARB) extra-virgin olive oil (OO) were monitored during frying by electronic nose (EN) and solid-phase microextraction–gas chromatography methodologies. Degradation of α- and γ-tocopherols was monitored by HPLC. Electronic nose data and volatile compounds were analysed at intervals of 15 min ( t ₁₅) during 60 min of frying ( t ₆₀). α- and γ-tocopherols were determined at intervals of 5 min ( t ₅) during 30 min of frying ( t ₃₀). Principal components analysis applied to EN data showed one component, PC₁ which accounted 96.6% of the total odour variation. SnO₂ sensors had a positive correlation with PC₁. ARA variety corresponding to frying t ₆₀ had the highest positive correlation with PC_1. Analysis of variance results for volatile compounds showed an increase on production for: 3-methyl butanal, n -pentanal, n -hexanal, n -heptanal and n -nonanal at 15 min of frying for ARB OO and at 30 min for ARA OO. α-tocopherol and γ-tocopherol showed an important decrease after the first 5 min of frying for ARB OO and at 15 min for ARA OO.     

EFFECTS OF ESCULETIN AS A LIPOXYGENASE INHIBITOR IN SOYBEAN EXTRACTS

This study was designed to determine the ability of esculetin to inhibit lipoxygenase activity in soybean. Lipoxygenase was extracted from ground soybean using 0.1 M Tris-HCl buffer (pH 8.6) and centrifuged at 3000 g for 15 min at 4C. The extract was mixed with different volumes of 0.05M esculetin solutions to provide 1.8 × 10⁻⁴M, 3.3 × 10⁻⁴M, 4.6 × 10⁻⁴M and 5.7 × 10⁻⁴M to final assay systems, respectively. Lipoxygenase activity decreased as the concentration of esculetin solution mixed in soybean extract increased. Mixing of 1.8 × 10⁻⁴M esculetin solution to soybean extract showed the least inhibition effect and was significantly different from that of 3.3 × 10⁻⁴M, 4.6 × 10⁻⁴M and 5.7 × 10⁻⁴M which inhibited LOX (lipoxygenase)'s activity of soybean extracts. In the study comparing the inhibition abilities for lipoxygenase activity among selected antioxidants, esculetin was significantly better than BHA (butylated hydroxyanisole) and α-tocopherol.     

α-, γ- and δ-Tocopherol Effects on Chlorophyll Photosensitized Oxidation of Soybean Oil

The effects of 0, 1.0, 2.0 and 4.0 (x 10^?3 M) α-, γ- or δ-tocopherol on chlorophyll b photosensitized oxidation of soybean oil in methylene chloride were studied by peroxide values and headspace oxygen. As concentrations of tocopherols increased, peroxide values decreased and headspace oxygen increased (P < 0.05). At 1.0 × 10^?3 M, α-tocopherol showed highest antioxidant effect, γ-tocopherol second and then δ-tocopherol. α- and -γ-Tocopherols had similar effects and δ-tocopherol had lower effect at 2.0 × 10^?3 M (P < 0.05). However, the three tocopherols were not different (P > 0.05) at 4.0 × 10^?3 M. α-Tocopherol quenched singlet oxygen to reduce the photosensitized oxidation of oil. The quenching rate constants of α-tocopherol were 2.7 × 10⁷M^?1sec^?1 by peroxide value and 2.6 × 10⁷ M^?1sec^?1 by headspace oxygen.     

Study of α-, γ-, and δ-tocopherols in the oxidative stability of lard

Effects of α-, γ-, and δ-tocopherols at 0, 50, 100, 250, 500, and 1,000 ppm on the oxidative stability of lard during storage in the dark at 55°C for 7 days were studied by determining headspace oxygen content and peroxide value. The headspace oxygen content of the lard without tocopherol addition decreased from 21.1 to 14.4% and the peroxide value increased from 2.5 to 15.4 meq/kg lard for 7 days. The headspace oxygen depletion and the peroxide value of lard decreased by the addition of α-tocopherol at 50 ppm, but significantly increased at 250, 500, or 1,000 ppm. As the concentration of γ-tocopherol or δ-tocopherol increased, the headspace oxygen depletion and the peroxide value decreased. α-Tocopherol at 250 ppm acted as a prooxidant in the autoxidation of lard. The concentrations of γ- or δ-tocopherol to be applied to improve the autoxidative stability of lard may be 100–250 ppm.     

CHANGES IN SIZE OF GAS CELLS IN DOUGH AND BREAD DURING BREADMAKING AND CALCULATION OF CRITICAL SIZE OF GAS CELLS THAT EXPAND

Microscopic observation showed that a group of small air cells entrained during the early stage of mixing is the original cause of cell structure of bread. At the beginning of fermentation, about 3 × 10⁸/m² gas cells with diameters between 3 × 10⁻⁶ and 8 × 10⁻⁴ m were entrained in the dough. The distribution curve of cell size was approximately normal on a logarithmic scale. During fermentation and proofing, a great portion of carbon dioxide was released into cells larger than about 10⁻⁴ m in diameter that was equivalent to a few percentages of total number of gas cells. After baking, gas cells smaller than 10⁻⁴ m in diameter were not observed and the total number of cells in baked bread reduced to about 10⁶/m² with diameters between 10⁻⁴ and about 5 × 10⁻³ m. The critical cell size to expand generally agreed with the calculated value using an equation, rc'= 3s/E (re': critical radius to expand, s: surface tension, E: elasticity), and cited value of s and E.     

Natural Antioxidants as a Component of an Egg Albumen Film in the Reduction of Lipid Oxidation in Cooked and Uncooked Poultry

ABSTRACT: Egg albumen coatings with natural antioxidants fenugreek, rosemary, and vitamin E were evaluated for their antioxidant activity in diced raw and diced cooked poultry breast meat. Coatings were applied using a vacuum tumbling and a boiling method. Coated samples and uncoated controls were evaluated for their malondialdehyde and reactive substances content as a measure of lipid oxidation via the thiobarbituric acid test. Cooked egg albumen-coated samples showed reactive compound changes of 1.56 × 10⁻⁷M for 4 d of refrigeration, compared to 1.59 × 10⁻⁶ M change in the cooked control. Raw control changed by 1.97 × 10⁻⁶M versus 1.67 × 10⁻⁶ M for raw rosemary coated samples. In raw and cooked studies EAS coating showed most effective against lipid oxidation.     

RHEOLOGICAL AND CHEMICAL PROPERTIES OF MOZZARELLA CHEESE

Dynamic viscoelastic parameters and chemical properties of Mozzarella cheese produced using a "no-brine" cheese making method with 3 different cooking temperatures (38, 41, and 44C) were determined. Samples were stored for 3 weeks at 4C before dynamic mechanical analysis at 22C. G', G" and tan δ were 5.8 – 6.4 × 10⁵ dyne/cm², 1.9 – 2.1 × 10⁵ dyne/cm², and 0.33 – 0.35, respectively, at 1% strain and 10 rad/s. The percentage of intact α_s-casein and β-casein were 38–40% and 33–35% of total protein in the cheese, respectively. The range of cooking temperatures used in this experiment had little effect on dynamic viscoelastic properties or the amount of intact protein for the cheese.     

MICROBIOLOGICAL QUALITY OF TRADITIONAL MARKET CURED FISH IN TANZANIA

The microbiological quality of six varieties of retail market traditionally cured fish in Morogoro, Tanzania was investigated over a five-month period. The fish were contaminated with bacteria and molds at levels of: total aerobes, 10⁶ - 1.7 × 10⁷ c.f.u/g; faecal coliforms, 1.1 × 10¹ - 2.5 × 10³ MPN/g; faecal streptococci, 1.4 × 10¹ - 1.3 × 10³ MPN/g; Staphylococcus aureus, 1.3 × 10³ - 8.6 × 10³ c.f.u/g; Aspergillus flavus group, 2.1 × 10¹ - 2 × 10² c.f.u/g of fish. Of faecal coliform, 45% of the isolates were Escherichia coli. Twenty-five percent of the S. aureus isolates were coagulase positive. Sixteen percent of A. flavus isolates were aflatoxigenic. Aflatoxin contamination ranged from O to 18.5 μg/kg of fish. Insect infestation by Dermestes spp. and mites was observed. The results of this preliminary study emphasize the importance of proper processing and handling offish in the tropics in order to safeguard public health .     

Rapid dye reduction tests for the determination of microbiological quality of meat

Rapid dye reduction tests have been developed to determine the quality of meat. Three chemical indicators, resazurin and two tetrazolium compounds, were used to correlate the microbial numbers and reduction times in meat samples. Twenty-five surface samples from sheep carcasses were subjected to each reduction test. Total viable counts given were obtained at 37°C. Resazurin reduction time was 90–120 min when the bacterial counts ranged from 1.5×10⁶ to 7.7×10⁶/cm². Samples showing bacterial counts between 1.5×10⁶ and 6.0×10⁶/cm² reduced tetrazolium (NBT) in 360–390 min whereas samples containing bacterial counts of 2.1×10⁶/cm² took 420–450 min to reduce iodophenyl nitrophenyl tetrazolium (INT) dye. Regression equations relating the number of organisms per cm² and reduction time were applied to predict the microbiological quality of meat samples from reduction time data. Among the three dyes, resazurin gave the lowest reduction time.     

BACTERIAL FORMATION OF HISTAMINE IN JACK MACKEREL (TRACHURUS SYMMETRICUS)

Peak histamine concentrations of 0.023, 0.031 and 0.027 g histamine/100 g muscle and maximal bacteria concentrations of 1.75, 1.59 and 0.423 g dry cells/100 g muscle were observed in muscles of jack mackerel stored at 25, 15 and 5C, respectively. Incubated fish homogenates suggest rate and transport limitations in histamine formation in muscle. The Mulchandani model predicted bacterial growth in muscle. The Luedeking and Piret expression fitted histamine formation in muscle; α values were 3.0 × 10⁻³, 1.23 × 10⁻² and 4.17 × 10⁻² g histamine/g dry cells, while β-values were 4.5 × 104, 8.0 × 10⁻⁵ and 0 g histamine/g dry cells × h at 25, 15, and 5C, respectively. The model predicts that jack mackerel could be stored from 4.5 to 5.5 days in ice, from 1 to 2 days at 15C and from 17 h to 2 days at 25C before fishmeal quality might be affected.     

Alginate- and gellan-based edible films for probiotic coatings on fresh-cut fruits

ABSTRACT:  Alginate- (2% w/v) or gellan-based (0.5%) edible films, containing glycerol (0.6% to 2.0%), N-acetylcysteine (1%), and/or ascorbic acid (1%) and citric acid (1%), were formulated and used to coat fresh-cut apple and papaya cylinders. Water vapor permeability (WVP) was significantly higher ( P < 0.05) in alginate films (0.30 to 0.31 × 10⁻⁹g m/Pa s m²) than in the gellan ones (0.26 to 0.27 × 10⁻⁹g m/Pa s m²). Addition of 0.025% (w/v) sunflower oil decreased WVP of gellan films (0.20 to 0.22 × 10⁻⁹ g m/Pa s m²). Water solubility of gellan and alginate films at 25 °C (0.47 to 0.59 and 0.74 to 0.79, respectively) and their swelling ratios (2.3 to 2.6 and 1.6 to 2.0, respectively) indicate their potential for coating high moisture fresh-cut fruits. Fresh-cut apple and papaya cylinders were successfully coated with 2% (w/v) alginate or gellan film-forming solutions containing viable bifidobacteria. WVP in alginate (6.31 and 5.52 × 10⁻⁹g m/Pa s m²) or gellan (3.65 and 4.89 × 10⁻⁹ g m/Pa s m²) probiotic coatings of papaya and apple, respectively, were higher than in the corresponding cast films. The gellan coatings and films exhibited better water vapor properties in comparison with the alginate coatings. Values > 10⁶ CFU/g B. lactis Bb -12 were maintained for 10 d during refrigerated storage of fresh-cut fruits, demonstrating the feasibility of alginate- and gellan-based edible coatings to carry and support viable probiotics on fresh-cut fruit.     

Antioxidant Properties of Edible Basidiomycete Phellinus igniarius in Submerged Cultures

ABSTRACT:  Dried mycelia and mycelia-free broths produced by submerged cultures of  Phellinus igniarius  under optimal culture conditions were extracted using methanol and hot water and investigated for antioxidant properties. Methanolic extracts from dried mycelia (MEM) and mycelia-free broth (MEB) showed significant antioxidant properties for all EC₅₀ values less than 10 mg/mL except for MEB in scavenging effects on DPPH radicals. Hot water extracts from dried mycelia (HWEM) were evidenced by their low EC₅₀ values (<10 mg/mL) to be effective in reducing power, chelating effect on ferrous ions, and scavenging effect on superoxide anions. α-tocopherol was mainly found in MEM and γ-tocopherol in MEB. Ascorbic acid and total flavonoids were abundant in methanolic extracts (MEM + MEB), whereas total phenols were rich in HWEM. An excellent correlation between contents of total phenols and EC₅₀ values was accomplished for antioxidant activity ( R ²= 0.996) and chelating ferrous ions ability ( R ²= 0.922).
Practical Application: In our paper, the products by submerged culture of  Phellinus igniarius  exhibited powerful antioxidant properties. Results told that extracts from fermenting products by  P. igniarius  might be good sources for antioxidant-related functional foods and pharmaceutical industries.     

QUANTITATIVE STUDIES OF PROTEOLYTIC AND LIPOLYTIC BACTERIA IN FROZEN MINCED BEEF AND HAMBURGER

Sixty frozen samples (30 hamburger patties and 30 minced meat) purchased from different retail markets in Ismailia, Egypt were examined for incidence of proteolytic and lipolytic bacteria. Mean values of proteolytic psychrophiles in the examined samples of hamburger and minced meat were 2×10⁵ and 6×10⁴, respectively. Lipolytic psychrophiles in hamburger were 8×10⁵ and 3×10⁵ in minced meat. Proteolytic mesophiles in hamburger and minced meat were 6×10⁵ and 5×10⁴, respectively. Mean values of lipolytic mesophiles were 5×10⁴ for hamburger and 5×10³ for minced meat. Proteolytic thermophiles in hamburger and minced meat were 3×10³ and 10³, respectively. Both proteolytic and lipolytic activities were exhibited by various bacteria that were identified.     

PARTIAL PURIFICATION AND CHARACTERIZATION OF α-GALACTOSIDASE FROM ASPERGILLUS ORYZAE

A crude extract of α-galactosidase obtained by fermenting Aspergillus oryzae on wheat bran was purified 35 fold by ethanol precipitation, gel filtration and ion-exchange chromatography. The final preparation was free of protease activity but contained invertase activity. The molecular weight of the enzyme was estimated as 64,000 daltons. The pH and temperature optima were 4.0 and 60°C, respectively. The enzyme was stable over the pH range 3–7.5 and at temperatures up to 55°C (pH 4.0). The K_m values for p-nitrophenyl α-Dgalactopyranoside (PNPG) and raffinose were 4.0 × 10⁻⁴M and 1 × 10⁻²M, respectively. Divalent cations were not required for activity. More than 80% of the oligosaccharides in soy milk were hydrolyzed after 3 h at 50°C using 0.113 PNPG units/mL milk.