Electron beam ionization induced oxidative enzymatic activities in pepper (Capsicum annuum L.), associated with ultrastructure cellular damages

Mature green pepper fruits (Capsicum annuum L.) were subjected to ionizing radiation, in the range of 1-7 kGy, with accelerated electrons. Ultrastructural changes by electron microscopy, and the activity of several oxidative metabolism-related enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaicol peroxidase (POX), and lipoxygenase (LOX), were determined in pericarp tissue just after the ionization treatment and during postionization storage at 7 degrees C followed by 3 days at 20 degrees C. Changes in oxidative stress during the ionization treatment was assessed by the accumulation of malondyaldehide (MDA), a lipid peroxidation product. The ionization induced modifications in the cell ultrastructure, a moderate separation of the plasma membrane from the cell wall being observed for all doses. At 5 and 7 kGy, peroxisomes were not detected and the structures of the chloroplast and vacuoles were seriously damaged. Lipid peroxidation and lipoxygenase activity increased with the ionization dose, staying constant and decreasing, respectively, during the storage period. Conversely, catalase, ascorbate peroxidase, and superoxide dismutase had lower values than in nonionized fruits and, in general, their values did not change or diminished slightly from the seventh day of storage. Peroxidase exhibited an increase in activity with the ionization dose, although these was not a linear relationship, with higher values at 3kGy. Ionization of pepper, especially at doses of 5 and 7 kGy, caused a significant oxidative damage in the fruit, since it increased oxidation and decreased the antioxidant enzymatic defense systems causing ultrastructural changes at cell level.     

Determination of rutin, catechin, epicatechin, and epicatechin gallate in buckwheat Fagopyrum esculentum Moench by micro-high-performance liquid chromatography with electrochemical detection

A simple and sensitive method has been developed for determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat (Fagopyrum esculentum Moench) flour and seeds by micro-high-performance liquid chromatography with electrochemical detection. Chromatography was performed using an octadecylsilica column, acetonitrile-water-formic acid (13:87:1, v/v/v) as a mobile phase, and an applied potential at +0.5 V vs Ag/AgCl. We found that Japanese buckwheat flour contains rutin (12.7 mg/100 g), catechin (3.30 mg/100 g), epicatechin (20.5 mg/100 g), and epicatechin gallate (1.27 mg/100 g). The relative standard deviations for rutin, catechin, epicatechin, and epicatechin gallate peak heights were less than 0.86% (n = 5). The detection limit of rutin was 0.86 ng/mL. Moreover, the present method was applied to the distribution analysis of these compounds in buckwheat seed. The embryo proper and cotyledons of a mature buckwheat seed contained rutin with the highest concentration as compared to other parts. This method is useful in determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat with a small amount of sample for quality control in the food industry.     

Extraction of rutin from buckwheat (Fagopyrum esculentumMoench) seeds and determination by capillary electrophoresis.

The content of the flavonoid rutin was determined in different milling fractions of buckwheat seeds and in buckwheat stems, leaves, and flowers. The extraction was performed by using a solvent containing 60% of ethanol and 5% of ammonia in water. The extracts were analyzed by capillary electrophoresis (running buffer of 50 mM borate (pH 9.3), 100 mM sodium dodecyl sulfate; determination at 380 nm). In bran fractions the concentration of rutin was 131-476 ppm, and in flour fractions 19-168 ppm. On average, about 300, 1000, and 46000 ppm of rutin were found in leaves, stems, and flowers, respectively. The results indicate that buckwheat could be an important nutritional source of flavonoids, especially in countries with a low mean daily flavonoid intake.     

Purification and characterization of lipase in buckwheat seed

To obtain basic information about enzymatic deterioration of buckwheat flour, triacylglycerol lipase (LIP; EC 3.1.1.3) was purified from buckwheat seed. The LIP consisted of two isozymes, LIP I and LIP II, and they were purified with purification folds of 60 and 143 with final specific activities of 0.108 and 0.727 mumol of fatty acid released per minute per milligram of protein at 30 degrees C using triolein as a substrate. Molecular weights were estimated to be 150 (LIP I) and 28.4 kDa (LIP I) by gel filtration and 171 (LIP I) and 26.5 kDa (LIP II) by SDS-PAGE. Optimal pHs of LIP activities were 3.0 (LIP I) and 6.0 (Lip II) using triolein as a substrate. Both LIP I and II reacted in the acidic pH range. Optimal temperatures were 30 (LIP I) and 40 degrees C (LIP II), and both LIP I and II were stable below 30 degrees C when p-nitrophenyl-laurate was used as a substrate. However, they were inactivated above 60 degrees C. On the other hand, when triolein was used as a substrate, optimal temperatures were 30 degrees C for both LIP I and II, and they retained 40% of their activity after a 4 h incubation of enzymes at 70 degrees C. LIP I and II had higher activity against triolein than monoolein or tri/monopalmitin. Most of the LIP activity was distributed in the embryo.     

Commercial scale pulsed electric field processing of tomato juice

Effects of commercial scale pulsed electric field (PEF) processing on the quality of tomato juice were studied and compared with those of thermal processing. Tomato juice was prepared by hot break at 88 degrees C for 2 min or by cold break at 68 degrees C for 2 min and then thermally processed at 92 degrees C for 90 s or PEF processed at 40 kV/cm for 57 micros. Thermally processed, PEF processed, and unprocessed control juices were packed into 50 mL sterilized polypropylene tubes in a sanitary glovebox and stored at 4 degrees C for 112 days. Both thermally and PEF processed juices showed microbial shelf life at 4 degrees C for 112 days. The lipoxygenase activities of thermally and PEF processed juices were 0 and 47%, respectively. PEF processed juice retained more ascorbic acid than thermally processed juice at 4 degrees C for 42 days (p < 0.05). No significant differences were observed in the concentration of lycopene, degrees Brix, pH, or viscosity between thermally and PEF processed juices during the storage (p > 0.05). Sensory evaluations indicated that flavor and overall acceptability of PEF processed juice were preferred to those of thermally processed juice (p < 0.05).     

Prevention of oxidative rancidity in rice bran during storage.

The effect of microwave heat on lipoxygenase (LOX) activity in rice bran under various storage conditions was examined. Raw rice bran from the long-grain variety Lemont was adjusted to 21% moisture content and heated in a microwave oven at 850 W for 3 min. Raw and microwave-heated rice bran samples were packed in zipper-top bags or vacuum packs and stored at room temperature (25 degrees C) or in the refrigerator (4-5 degrees C) for 16 weeks. Samples were analyzed for LOX activity at 4-week intervals. LOX activity did not significantly change from its initial value at week 0 for zipper-top and vacuum-packed samples while stored at 4-5 degrees C for 12 weeks, but decreased at week 16. Vacuum packing did not show a significant impact on LOX activity during 16 weeks of storage. Microwave-heated samples stored in the refrigerator did not show significant change in LOX activity for up to 12 weeks but showed a significant (p < 0. 05) decrease at 16 weeks. Results showed that oxidative rancidity of rice bran could be prevented by microwave heating the samples, packing in zipper-top bags, and storing at 4-5 degrees C for up to 16 weeks.     

Stability of dried and intermediate moisture tomato pulp during storage

Commercial tomato pulp was air-dried to two final moisture contents in order to obtain intermediate moisture pulp (IMP, 23% moisture) and dried pulp (DP, 9% moisture). IMP and DP were stored at 4, 20, and 37 degrees C for approximately 5 months; periodically samples were analyzed to evaluate heat and oxidative damage by measurement of color changes, total phenolics, rutin, lycopene and furosine concentrations, and antioxidant activity of the lipophilic extract. IMP and DP, despite different drying degree, had similar antioxidant activity; in fact, whereas lycopene was stable to drying treatments, ascorbic acid was totally degraded in both products. During storage, IMP and DP showed different behavior: IMP was more sensitive to degradation than DP, especially with regard to lycopene, rutin, and antioxidant activity degradation and to nonenzymatic browning. Effects of storage temperature varied with regard to different parameters: variations in rutin, furosine, and color indices were higher in products stored at higher temperatures, while lycopene and antioxidant activity of the lipophilic fraction were maximally degraded in products stored at 4 degrees C.     

Distribution of Vitamin E, squalene, epicatechin, and rutin in common buckwheat plants (Fagopyrum esculentum Moench)

Buckwheat leaves and young parts of the plant are consumed in some countries as a vegetable. Green flour, obtained by milling of the dried plants, is used as a natural food colorant. The distribution of vitamin E, squalene, epicatechin, and rutin (as the most important antioxidants) within buckwheat plants, as well as changes of their content within leaves during the growing season, were determined by GC-MS and HPLC analyses. alpha-Tocopherol was found as the main component of vitamin E in all parts of the plant; epicatechin and squalene were also detected. For the use of buckwheat as an antioxidant source in the human diet, the most suitable part of the plants seems to be the leaves and the flowers at the stage of full flowering due to the considerable amounts of rutin and epicatechin. alpha-Tocopherol content correlates positively with temperature, drought, and duration of solar radiation. Certain differences appear among varieties of buckwheat, especially in their squalene and rutin contents.     

Oxido-Reductases and Lipases as Dough-Bleaching Agents

To replace benzoyl peroxide as a bread dough-bleaching agent, pure and commercial oxido-reductases (peroxidases, catalases, glucose oxidases, lipoxygenase, and laccase) were screened based on degradation of β-carotene in a liquid system (5 μg of β-carotene/mL of 0.1M citrate phosphate buffer at pH 5.5 or 6.5) or dough. Peroxidases had the best bleaching activity; some catalases also showed bleaching potential in a liquid system but not in bread dough, suggesting that screening enzymes in liquid media has limited application for dough. In 100 g of flour, combinations of peroxidase (3,000 U), lipase (815–1,630 U), and linoleic acid (0–300 mg) completely bleached bread dough.     

Fruit antioxidant activity, ascorbic acid, total phenol, quercetin, and carotene of Irwin mango fruits stored at low temperature after high electric field pretreatment

Greenhouse-grown tree ripe (TR) and mature green (MG) mangoes (cv. Irwin) were exposed to high electric field treatment before 20 and 30 days of storage at 5 degrees C. MG fruits were allowed to ripen at room temperature after low-temperature storage. Fruit physical quality attributes, ascorbic acid, carotene, quercetin, total phenols, and antioxidant capacity were estimated before and after the storage period. Antioxidant capacity of fruit juice was estimated using the ferric reducing antioxidant power (FRAP) assay. Fruit firmness decreased significantly during storage. Titratable acidity decreased 20 days after storage. Total soluble solids did not change during storage. Antioxidant capacity of fruits remained unchanged up to 20 days of storage period and decreased thereafter. Total phenol and carotenes increased during storage. Antioxidant capacity of fruits was significantly correlated only to ascorbic acids. Peel color and carotenes were higher in TR fruits, whereas titratable acidity and firmness were higher in MG fruits. There was no significant difference in other parameters between the stages of picking. Electric field pretreatment affected the respiration and antioxidant capacity of TR fruits and did not have any significant affect on other parameters. TR mangoes of cv. Irwin are more suitable for low-temperature storage and can be successfully stored for up to 20 days at 5 degrees C without any significant losses in functional properties and quality attributes.     

Optimal conditions for phytate degradation, estimation of phytase activity, and localization of phytate in barley (cv. Blenheim)

Using a multivariate experimental design, optimal conditions for phytate degradation were found to be pH 4.8 and 57 degrees C in barley flour (cv. Blenheim) and pH 5.2 and 47 degrees C in a crude extracted phytase from barley. Three methods for measuring phytase activity in raw and hydrothermally processed barley were compared. Incubation at pH 5 and 55 degrees C for 60 min did not give significantly different results (p > 0.05), whereas incubation at pH 5 and 50 degrees C for 10, 20, 30, and 60 min gave significantly different results (p < 0.001) between methods. The change in microstructure of phytate globoids during hydrothermal processing showed that the degradation was highest in the scutellum cells and less in the aleurone layer.     

Partial Characterization of Buckwheat (Fagopyrum esculentum) Starch

Laboratory-isolated buckwheat (Fagopyrum esculentum) starch was compared to commercial corn and wheat starches. Buckwheat starch granules (2.9–9.3 μm) were round and polygonal with some holes and pits on the surface. Buckwheat starch had higher amylose content, waterbinding capacity, and peak viscosity, and it had lower intrinsic viscosity when compared with corn and wheat starches. Buckwheat starch also showed restricted swelling power at 85–95°C and lower solubility in water at 55–95°C and was more susceptible to acid and enzymatic attack. Gelatinization temperatures, determined by differential scanning calorimetry, were 61.1–80.1°C for buckwheat starch compared to 64.7–79.2°C and 57.1–73.5°C for corn and wheat starches, respectively. A second endotherm observed at 84.5°C was an amylose-lipid complex attributed to the internal lipids in buckwheat starch, as evidenced by selective extraction. The retrogradation of buckwheat, corn, and wheat starch gels was examined after storage at 25, 4, and -12°C for 1–15 days. In general, buckwheat starch retrogradation was slower than that of corn and wheat starch, but it increased as storage time increased, as did that of the other starch pastes. When the values of the three storage temperatures were averaged for each storage period analyzed, buckwheat starch gels showed a lower percentage of retrogradation than did corn and wheat starch gels. Buckwheat starch also had a lower percentage of water syneresis when stored at 4°C for 3–10 days and had better stability to syneresis after three freeze-thaw cycles at -12 and 25°C.     

Extrusion Properties and Cooking Quality of Spaghetti Containing Buckwheat Bran Flour

The effect of hydration level on processing properties and the effects of hydration level, concentration of buckwheat bran flour and drying temperature on the physical and cooking quality of spaghetti were determined. Specific mechanical energy transferred to the dough during extrusion decreased 69% for semolina and 79% for semolina containing 30%, w/w, buckwheat bran flour, as hydration level increased 29–32% absorption. Little or no postdrier checking occurred in spaghetti made from semolina or spaghetti containing buckwheat bran flour when dried at high (70°C) or ultrahigh temperature (90°C). When dried at low temperature (40°C), tolerance to postdrier checking of spaghetti decreased as buckwheat bran flour increased 0–30% (w/w). Hydration level before extrusion did not affect cooking loss of spaghetti made from semolina. However, cooking loss was greater from spaghetti made with semolinabuckwheat bran flour that was hydrated to 32% compared with 29–31% absorption. Cooked firmness of spaghetti containing buckwheat bran flour decreased from 0.588–0.471 Nm as hydration increased from 29–32% absorption. Cooking loss was lower and cooked firmness was greater when spaghetti containing buckwheat bran flour was dried at ultrahigh than at low temperature.     

Kinetics of carotenoids degradation during the storage of einkorn (Triticum monococcum L. ssp. monococcum) and bread wheat (Triticum aestivum L. ssp. aestivum) flours

To evaluate the effect of storage temperature, the degradation kinetics of carotenoids in wholemeal and white flour of einkorn cv. Monlis and bread wheat cv. Serio, stored at -20, 5, 20, 30, and 38 degrees C, was assessed by normal-phase high-performance liquid chromatography. In Monlis, the carotenoids content (8.1 and 9.8 mg/kg for wholemeal and white flour, respectively) was 8-fold higher than in Serio (1.0 and 1.1 mg/kg). Only lutein and zeaxanthin were detected in bread wheat, while significant quantities of (alpha and beta)-carotene and beta-cryptoxanthin were observed in einkorn. Carotenoids degradation was influenced by temperature and time, following first-order kinetics. The degradation rate was similar in wholemeal and white flour; however, loss of lutein and total carotenoids was faster in Serio than in Monlis. The activation energy E(a) ranged from 35.2 to 52.5 kJ/mol. Temperatures not exceeding 20 degrees C better preserve carotenoids content and are recommended for long-term storage.     

Optimization of Temperature,Time, and Lactic Acid Concentration to Inactivate Lipoxygenase and Lipase and Preserve Phytase Activity in Barley (cv. Blenheim) During Soaking

A barley-soaking process was studied to find conditions that inactivate the prooxidative enzyme lipoxygenase and the lipolytic enzyme lipase but preserve phytase activity to develop possible procedures for production of barley products with potentially high mineral bioavailability and good oxidative stability. Lactic acid concentration, temperature, and soaking time were studied. The study was done using a multivariate experimental design. Lactic acid concentration varied between 0 and 1%, temperature varied between 45 and 70°C, and soaking time varied between 30 and 120 min. Although conditions under which lipoxygenase was inactivated were found, total inactivation of lipase was not obtained. Total lipoxygenase inactivation with <20% remaining lipase activity and >60% remaining phytase activity was reached after soaking in 1% lactic acid at suitable time-temperature combinations of 70–110 min and 53–58°C.     

Effect of Exogenous Lipase on Dough Lipids During Mixing of Wheat Flours

In control dough, endogenous wheat lipase was inactive, because the triacylglycerol (TAG), 1,2-diacylglycerol (DAG_1,2), and 1,3-diacylglycerol (DAG_1,3) fractions of nonpolar lipids were not affected by mixing. Conversely, the free fatty acid (FFA) and monoacylglycerol (MAG) fractions decreased, mainly due to the oxidation of polyunsaturated fatty acids (PUFA) catalyzed by wheat lipoxygenase. Addition of exogenous lipase to flour (15 lipase units [LU] per gram of dry matter) resulted in substantial modification of nonpolar lipids during dough mixing. Due to the 1,3 specificity of the lipase used in this experiment, the TAG and DAG_1,3 fractions decreased, whereas the MAG and FFA fractions increased. The DAG_1,2 fraction increased at the beginning of mixing and decreased after 40 min of mixing. Moreover, part of the PUFA released by lipase activity was oxidized by wheat lipoxygenase, resulting in major losses of PUFA. Conversely, the net content of the saturated and monounsaturated fatty acids (SMUFA) remained constant, because the free SMUFA content increased primarily at the expense of the esterified forms. For a constant mixing time of 20 min, increasing the amount of lipase added to dough (from 2.5 to 25 LU/g of dry matter) resulted in a linear decrease in the TAG fraction and a linear increase in the SMUFA content in the FFA fraction. At the same time, the PUFA content of the FFA fraction increased only for additions of lipase to flour of >5 LU/g of dry matter, due to partial oxidation by wheat lipoxygenase.     

gamma-Radiation influences browning, antioxidant activity, and malondialdehyde level of apple juice

Apple juice was gamma-irradiated at 5 degrees C at doses ranging from 0 to 8.9 kGy and then stored at 5 degrees C for 15 days. Ionizing radiation reduced the browning of apple juice and increased antioxidant activity measured by the ferric-reducing antioxidant power (FRAP) assay. The magnitude of changes increased with radiation dose. The level of malondialdehyde (MDA) measured using the thiobarbituric acid reactive substrates assay increased at radiation doses above 2.67 kGy. The browning of irradiated juices increased during storage at 5 degrees C, but the irradiated juices were still lighter than controls at the end of storage. Differences in FRAP values disappeared during early periods of storage while higher MDA levels were observed in irradiated samples during most of the storage period. Elimination of suspended matter from apple juice did not alter irradiation-induced changes in browning, FRAP, or MDA formation. As compared to irradiation conducted at 5 and 20 degrees C, treatment at -15 degrees C was less effective in reducing browning and in increasing MDA formation but elevated FRAP values. The exclusion of oxygen from juices did not affect the reduction in browning due to irradiation but promoted the increase in FRAP values and decreased the irradiation-induced MDA formation.     

Influence of 1-methylcyclopropene on ripening,storage life,and volatile production by d'Anjou cv. pear fruit

d'Anjou cv. pear fruit (Pyrus communis L.) exposed at harvest to 0, 0.42, 4.2, or 42 micromol m(-)(3) 1-methylcyclopropene (1-MCP) for 12 h at 20 degrees C were stored at 1 degrees C for up to 8 months. After storage, half of the fruit was continuously exposed to ethylene (0.45 or 4-18 mmol m(-)(3)) for 7 days at 20 degrees C. All fruit treated with 1-MCP had lower respiration and ethylene production compared to untreated controls. Fruit quality changes were delayed following 1-MCP treatment, as was development of superficial scald and peel yellowing. The duration of 1-MCP-induced responses was dependent on 1-MCP treatment concentration. When 1-MCP-treated fruit began to ripen, softening and production of volatile compounds proceeded similar to that of untreated fruit. Post-storage ethylene exposure did not consistently stimulate ripening of fruit previously treated with 1-MCP. Efficacy of ethylene treatment depended on 1-MCP concentration and storage duration.     

Accumulation of Oxygenated Fatty Acids in Oat Lipids During Storage

Oxygenated fatty acids were identified in oat grain by gas chromatography‐mass spectrometry. We hypothesized that most of these were the results of lipoxygenase activity. This hypothesis was tested by measuring concentrations of these compounds after hydrothermal treatments and storage of oat groats or oat flour for 22 weeks at 37°C and 65% relative humidity. Steam treatments inactivated lipases, whereas roasting at 106°C did not. Free fatty acids accumulated quickly in untreated or roasted flour, but not in steamed flour or groats. A total of six hydroxy and epoxy fatty acids were identified. Oxidized fatty acids were found in both esterified lipids and free fatty acids, indicating that lipase action was not necessary for lipid oxidation. More oxidation products were found in flour than in groats, and less were found in the steamed treatments. Lipoperoxygenase appeared to be involved in the formation of oxidation products, although nonenzymatic mechanisms may also operate. Hydroxy‐fatty acids are associated with strongly bitter flavors and are undesirable. Results indicate the importance of enzyme inactivation before storage of processed oat products.     

Staling of Bread as Affected by Waxy Wheat Flour Blends

Crumb softness and improved shelf life of bread is often achieved by incorporating expensive shortenings in the formulation. We hypothesized that similar results could be achieved by blending bread wheat flour with waxy (low amylose) durum wheat flour. White pan bread was baked from 10, 20, and 30% waxy durum wheat flour composites and evaluated for loaf volume and crumb firmness over a period of 0, 3, and 5 days. The loaf volumes were not affected by the waxy flour blends. However, as staling progressed over 3–5 days, significant firming of crumb was observed in the control sample compared with loaves containing waxy flour. The firmness was inversely proportional to the level of waxy flour used in the blend. A 20% waxy wheat flour blend was optimal in retarding staling while producing bread quality comparable with the control. It was further established that bread made with 20% waxy flour gave lower firmness values after 5 days of storage in comparison to bread made with 3% shortening. These results suggest that 20% waxy wheat flour could substitute for use of shortening to achieve desirable crumb softness and to retard staling upon storage.