Purification and characterization of novel proteinase inhibitors from dried figs

Two types of the natural organic matter P and B were isolated from dried figs by gel permeation and high-performance liquid chromatography. The characterizations of their molecular structures were also performed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and infrared absorption spectrometry. As a result, these samples were revealed to inhibit the serine and cysteine proteinases chymotrypsin and papain (K(i) = 10(-)(6)-10(-)(4) M). The optimal inhibitory pH values of the P and B samples were observed to be approximately 5.5 and 5.0, respectively. The analyses of their UV-vis absorption spectra and infrared absorption spectra indicated that they would be a kind of humic substance. The mass spectrometry analyses showed that they possessed relatively small heterogeneous molecules with molecular masses around 692, 845, and 1389 Da for the P sample and around 551, 704, and 909 Da for the B sample.     

基于机器视觉和近红外光谱技术的杏干品质无损检测

干果品质直接影响其市场销售。该研究以杏干为对象探讨用机器视觉和近红外光谱技术快速无损检测干果内外品质的方法。拍摄杏干4个不同位置的彩色图像,用基于区域骨架化的填充法分割杏干,提取每种角度下的面积。从100个正常杏干样本中随机挑选75个为校正集,25个为预测集,用多元线性回归对杏干的实际质量和4个面的面积建模,得到校正集和预测集相关系数分别为0.9374和0.9307,杏干质量分级的准确率为90%。提出用基于平均灰度的区域增长法提取杏干缺陷,缺陷检测的准确率为84.5%。采用SNV对杏干近红外光谱进行预处理,然后分别采用偏最小二乘法(PLS)、向后区间偏最小二乘法(biPLS)及联合区间偏最小二乘法(siPLS)建立杏干糖度预测模型。结果表明,当全光谱范围被划分为22个子区间,优选出区间[17、2、3、9、20、13、7、18、15、11、6],主因子数为10时建立的biPLS糖度模型性能最好。其校正集相关系数和校正均方根误差分别为0.8983和1.23,预测集相关系数和预测均方根误差分别为0.8814和1.46。研究表明,机器视觉结合近红外光谱技术能对杏干内外品质进行综合检测,也可为其他干果的品质检测提供借鉴。     

Antioxidant capacity and phenolic content of grapes, sun-dried raisins, and golden raisins and their effect on ex vivo serum antioxidant capacity

Grapes and raisins provide phenolic antioxidants, which contribute to their potential health benefits. The objectives of this study were to compare the antioxidant capacity and phenolic content of green Thompson seedless grapes (the most common variety of grapes consumed in the United States), sun-dried raisins, and golden raisins (both produced from Thompson seedless grapes) and to observe the effects of their consumption over 4 weeks in 15 healthy human males with a cross-over design. The oxygen radical absorbance capacity (ORAC) (positive statistical significance for grapes after 2 weeks and golden raisins after 3 weeks), serum oxidation (positive statistical significance for golden raisin lag time after 4 weeks), total phenolics (no significant effects), and C-reactive protein (no significant effects) were monitored. Immediately postconsumption, there were some significant nonpositive changes. It is hypothesized that these negative results may be explained by postprandial oxidation, a known effect after carbohydrate consumption. Golden raisins had the highest antioxidant capacity and phenolic content. The consumption of a serving of grapes or raisins each day, in addition to a typical diet, may not be sufficient to overcome postprandial oxidation when consumed with other high carbohydrate foods but may have beneficial antioxidant effects over time.     

Effects of exogenous propylene on softening, glycosidase, and pectinmethylesterase activity during postharvest ripening of apricots

Apricots (Prunus armeniaca L. cv. Boccuccia spinosa) picked at the commercial ripening stage [soluble solids content (SSC) 12.6%] were left to reach full ripening in continuously humidified air at 20 degrees C. Changes in the rate of ethylene production, firmness, soluble solids concentration, and titratable acidity were measured. The alpha-D- and beta-D-glucosidases, alpha-L-arabinofuranosidase, alpha-D- and beta-D-galactosidases, beta-D-xylosidase, and alpha-D-mannosidase activities were assayed. To evaluate the influence of ethylene on glycosidase activity, propylene (500 microL x L(-1)) was applied to apricots for 24 and 48 h. In apricots ripened in air, ethylene production increased on the first day and exhibited a typical climacteric pattern. Good edible quality was reached in 5 days when SSC was at least 14% and acidity was between 1.1 and 1.2% (% malic acid). During postharvest ripening, alpha-L-arabinofuranosidase activity increased from 1.9 to 11.6 nkat until day 7. alpha-D-Galactosidase, alpha-D-mannosidase, and beta-D-galactosidase activity increased continuously but at a lower rate. beta-D-Xylosidase activity also increased, but the level of activity was lower than the other glycosidases assayed. Pectinmethylesterase (PME) decreased during the postharvest ripening, and propylene enhanced this pattern, by stimulating ethylene production. Even the activities of alpha-L-arabinofuranosidase, beta-D-xylosidase, alpha-D-mannosidase, and beta-D-galactosidase were greatly stimulated by the propylene treatment, which consequently induced rapid softening of the fruits.     

Changes in free amino acid and sugar levels of dried figs during aflatoxin B1 production by Aspergillus flavus and Aspergillus parasiticus

An aqueous slurry of gamma-irradiated sterilized dried figs was inoculated with toxigenic strains of Aspergillus flavus and Aspergillus parasiticus. During incubation at 28 degrees C, pH, fructose, glucose, and free amino acids were determined by high-performance liquid chromatography and liquid chromatography (LC)/mass spectrometry, respectively, over 13 time points (1-20 days). At the same 13 time points using a LC/time-of-flight mass spectrometry screening method, aflatoxin B 1 and other secondary metabolites were simultaneously monitored. During the course of incubation, the pH significantly decreased and aflatoxin B 1 formation correlated with a reduction in proline content for both fungi. Of the 22 free amino acids that were monitored, only proline and cystine were found to be critical in supporting aflatoxin production. Levels of fructose and glucose steadily declined during incubation, until glucose was almost exhausted after 21 days. These time-course experiments confirmed the need for carbon and nitrogen sources for aflatoxin production in dried figs, and the favorable composition of figs as a fungal growth medium.     

Fatty acid, triacylglycerol, phytosterol, and tocopherol variations in kernel oil of Malatya apricots from Turkey

The fatty acid, sn-2 fatty acid, triacyglycerol (TAG), tocopherol, and phytosterol compositions of kernel oils obtained from nine apricot varieties grown in the Malatya region of Turkey were determined ( P<0.05). The names of the apricot varieties were Alyanak (ALY), Cataloglu (CAT), C?loglu (COL), Hacihaliloglu (HAC), Hacikiz (HKI), Hasanbey (HSB), Kabaasi (KAB), Soganci (SOG), and Tokaloglu (TOK). The total oil contents of apricot kernels ranged from 40.23 to 53.19%. Oleic acid contributed 70.83% to the total fatty acids, followed by linoleic (21.96%), palmitic (4.92%), and stearic (1.21%) acids. The s n-2 position is mainly occupied with oleic acid (63.54%), linoleic acid (35.0%), and palmitic acid (0.96%). Eight TAG species were identified: LLL, OLL, PLL, OOL+POL, OOO+POO, and SOO (where P, palmitoyl; S, stearoyl; O, oleoyl; and L, linoleoyl), among which mainly OOO+POO contributed to 48.64% of the total, followed by OOL+POL at 32.63% and OLL at 14.33%. Four tocopherol and six phytosterol isomers were identified and quantified; among these, gamma-tocopherol (475.11 mg/kg of oil) and beta-sitosterol (273.67 mg/100 g of oil) were predominant. Principal component analysis (PCA) was applied to the data from lipid components of apricot kernel oil in order to explore the distribution of the apricot variety according to their kernel's lipid components. PCA separated some varieties including ALY, COL, KAB, CAT, SOG, and HSB in one group and varieties TOK, HAC, and HKI in another group based on their lipid components of apricot kernel oil. So, in the present study, PCA was found to be a powerful tool for classification of the samples.     

Phenolic profiles of raw apricots, pumpkins, and their purees in the evaluation of apricot nectar and jam authenticity

The possibility of proving the undeclared addition of pumpkin puree in apricot nectars and jams has been investigated by using the phenol compound fingerprint and sensory evaluation. The cheaper pumpkin admixtures in apricot nectars and jams could not be detected by the sensory evaluation, particularly if present in quantities of <15%. The lower admixtures of pumpkin puree in apricot nectars and jams could be detected by the presence of syringic acid, a phenolic compound characteristic of the investigated pumpkins (Cucurbita pepo cv. Gleisdorff and Table Gold, Cucurbita maxima cv. Turkinja, and Cucurbita moschata cv. Argenta). Syringic acid was isolated from pumpkin puree and determined by using HPLC with diode array detection. By using the phenolic profile, undeclared pumpkin admixture (> or =5%) in the apricot nectars and jams could be proven.     

Residue levels and storage responses of nectarines, apricots, and peaches after dip treatments with fludioxonil fungicide mixtures

Mature apricots (Prunus armeniaca), nectarines [Prunus persica var. nectarine (Ait.)], and peaches [P. persica (L.) Batsch.] were subjected to a 2 min dip treatment with warm water at 48 degrees C or with fludioxonil (FLU) at 100 mg L-1 and 20 degrees C or at 25 mg L-1 FLU and 48 degrees C and then stored at 5 degrees C and 90-95% relative humidity (RH) for 1 week plus 1 additional week at 18 degrees C and approximately 80% RH. Fruit residue uptake was determined as a function of fungicide concentration, dip temperature, treatment time (only on nectarines), and fruit storage conditions. FLU residue level was closely related to fungicide concentration and treatment temperatures and was dependent on fruit species. FLU residues showed great persistence over both storage and shelf life. Fruit dipping in water at 48 degrees C effectively reduced decay development in cvs. 'May Grand' nectarines and 'Pelese' apricots but was ineffective in cvs. 'Red Top' and 'Sun Crest' nectarines during 7 days of storage compared with nontreated fruit. Decay rates in cvs. 'Glo Haven' peaches and 'Fracasso' apricots were very low in fruit dipped in water at both 20 and 48 degrees C. Fungicide treatments at 20 and 48 degrees C resulted in the total or almost total suppression of decay in all cultivars. During shelf life, fruit became very prone to decay, averaging 25.7-100% depending on the cultivar. Fruit dipping in hot water effectively reduced decay in 'Pelese' and 'Fracasso' apricots, 'Sun Crest' peaches, and 'May Grand' nectarines as compared to control, but was ineffective in 'Glo Haven' and 'Red Top' peaches. Fungicide treatments at 20 degrees C were more effective than hot water in most cultivars. The combination of FLU with water at 48 degrees C further improved the fungicide performance. Indeed, reduced levels (a fourth) of active ingredient were required to achieve a control of decay comparable to that for treatment at 20 degrees C. Residue levels in fruit after treatment with 100 mg L-1 FLU at 20 degrees C or with 25 mg L-1 FLU at 48 degrees C averaged approximately 0.6-2 mg kg-1, which were notably lower than the maximum residue limit (5 mg kg-1) allowed in the United States for stone fruit.     

Anti-inflammatory potential of flavonoid contents from dried fruit of Crataegus pinnatifida in vitro and in vivo

The dried fruits of Crataegus pinnatifida, a local soft drink material and medical herb, demonstrated antioxidant effect in a previous study. The present study investigates the anti-inflammatory potential of flavonoid contents from dried fruit of C. pinnatifida (CF-Fs). The preliminary investigation showed that CF-Fs (0.25-0.75 mg/mL) decreased the release of PGE2 and nitric oxide as induced by lipopolysaccharide (LPS, an endotoxin) in macrophage RAW 264.7 cells. The in vivo assay showed that pretreatment of rats with CF-Fs (50-200 mg/kg dosed by gavage) for 5 days significantly decreased the serum levels of the hepatic enzyme markers alanine aminotransferase and aspartate aminotransferase induced by the 6-h treatment with LPS (i.p.; 5 mg/kg). Histopathological evaluation of the rat livers revealed that CF-Fs reduced the incidence of liver lesions such as neutrophil infiltration and necrosis induced by LPS. Furthermore, it was found that pretreatment with CF-Fs decreased the hepatic expression of iNOS and COX-2 induced by LPS in rats. These results demonstrate that CF-Fs present anti-inflammatory potential in vitro and in vivo and that they may play a role in hepatoprotection.     

Influence of ethylene inhibition by 1-methylcyclopropene on apricot quality,volatile production,and glycosidase activity of low- and high-aroma varieties of apricots

Apricots of two varieties, Ceccona with strong aroma and San Castrese with low aroma but good firmness, were treated with 1 microL L(-)(1) 1-methylcyclopropene (1-MCP) for 12 h at 20 degrees C and then kept for shelf life at 20 degrees C and 85% relative humidity. 1-MCP treatment strongly inhibited ethylene production in apricots of both varieties, and softening was delayed. Fruit softening started before the rise of ethylene in air-treated apricots, which softened even when the rise of ethylene production was inhibited by 1-MCP. The softening reduction was more significant in Ceccona apricots than in San Castrese. Pectinmethylesterase (PME) activity declined in Ceccona fruit regardless of the treatment; in San Castrese, PME of air-treated fruit slightly increased, whereas in 1-MCP-treated apricots the activity declined. alpha-d-Galactosidase (alpha-gal) and beta-d-galactosidase (beta-gal) activities in Ceccona apricot were significantly reduced by 1-MCP treatment, whereas in San Castrese apricot no difference in activities was observed between air- and 1-MCP-treated fruit. The pattern of beta-d-xylosidase (xyl) activity in San Castrese apricot was similar to that of beta-gal, showing a peak on day 4 without difference between treatments. alpha-d-Mannosidase (alpha-man) activity of air-treated apricots of both varieties rose slightly, and 1-MCP treatment decreased the enzyme activity in both varieties. alpha-d-Glucosidase (alpha-glu) decreased in air-treated apricots in both varieties, and 1-MCP maintained higher activity in Ceccona fruit but not in San Castrese. Acidity decreased during postharvest ripening regardless of the treatment, whereas soluble solids content (SSC) increased in Ceccona apricot and slightly diminished in San Castrese ones without any effect by 1-MCP treatment. 1-MCP did not show any effect on apricot color; in contrast, it affected the volatiles profile, especially in Ceccona apricot, reducing the synthesis of lactones and promoting the rise of terpenols.     

Consistency of polyphenol oxidase (PPO) thermostability in ripening apricots (Prunus armeniaca L.): evidence for the presence of thermostable PPO forming and destabilizing mechanisms in apricots

Destabilization of thermostable polyphenol oxidase (TS-PPO) during the ripening of peaches has been previously shown (Yemenicio?lu, A.; Cemero?lu, B. Tr. J. Agric. For. 1998, 22, 261-265). This work studied the effect of ripening on thermal stability of apricot PPO for three different cultivars. Kabaa?i cultivar contained thermolabile PPO, whereas TS-PPO appeared in Hacihalilo?lu and Catalo?lu cultivars. The TS-PPO showed biphasic inactivation curves, and its D and z values between 60 and 90 degrees C varied in the ranges of 357-1.12 min and 11.9-12.7 degrees C, respectively. In Hacihalilo?lu cultivar the TS-PPO was very consistent and existed at all stages of ripening, whereas in Catalo?lu cultivar it appeared only at the half-ripe stage. The loss of consistent TS-PPO in Hacihalilo?lu apricots after partial purification by acetone precipitation and DEAE-cellulose chromatography suggested the non-covalent nature of its stabilization. The main purified fractions (F1 and F2) showed monophasic inactivation curves with similar thermal inactivation parameters (z(F1) = 10.4 degrees C, z(F2) = 10.1 degrees C). However, their kinetic properties against catechol (K(mF1) = 61 mM, K(mF2) = 122.7 mM) and substrate specificities were considerably different. The results of this study showed the presence of TS-PPO forming and destabilizing mechanisms in apricots. Further studies are needed for the solution of these mechanisms and to develop some new strategies that may be utilized by molecular techniques for a planned production of apricot cultivars provided with heat labile but normal PPO activity.     

Identification and occurrence of beta-carboline alkaloids in raisins and inhibition of monoamine oxidase (MAO)

Monoamine oxidase (MAO) is a mitochondrial enzyme involved in the oxidative catabolism of neurotransmitters and xenobiotic amines, including vasopressor and neurotoxic amines, and a current target for antidepressant and neuroprotective drugs. Raisin extracts and homogenates exhibited reversible in vitro inhibition of MAO isozymes, particularly MAO-A, suggesting the presence of MAO-inhibiting substances. Chromatographic and spectrometric studies showed the occurrence of aromatic beta-carboline alkaloids in raisins, and norharman and harman were identified as the key contributors to MAO inhibition. On average, harman ranged from 6 to 644 ng/g and norharman from 2 to 120 ng/g. Several technological variables appeared to determine the presence of these compounds in raisins. Dark-brown raisins (i.e., sun-dried) contained much higher levels than golden raisins. Tetrahydro-beta-carboline-3-carboxylic acid compounds that are direct precursors of aromatic beta-carbolines were also identified in raisins and appeared in a higher amount, reaching up to 50 microg/g. beta-Carbolines were isolated from raisins and acted as good competitive inhibitors of MAO-A (harman) and MAO-B (norharman) isozymes. These results suggest that beta-carboline alkaloids and perhaps raisins containing a high level of beta-carbolines might exhibit potential activity as MAO inhibitors. The results also show that some raisins can be a source of dietary exposure to bioactive beta-carbolines.     

Occurrence of fumonisins B(2) and B(4) in retail raisins

Concerns that raisins may be contaminated by fumonisins stem from the persistent occurrence of Aspergillus niger spores on raisins and the recent discovery of fumonisin production by A. niger on grapes, which leads to the widespread occurrence of fumonisin B(2) in wine. This study presents an LC-MS/MS survey of fumonisins in retail raisins. In 10 of 21 brands collected in Denmark, Germany, and The Netherlands, fumonisins B(2) and B(4) were detected at levels up to 13 and 1.3 μg/kg, respectively. Only fumonisin B(2) has been detected in wine, so the presence of fumonisin B(4) in raisins suggests that the fumonisins are produced mainly during the drying process concomitant with the decreasing water activity. Analysis of multiple packages from one manufacturer showed a 3-fold package-to-package variation, suggesting that a few raisins per package are contaminated.     

Determination of total N-nitroso compounds and their precursors in frankfurters, fresh meat, dried salted fish, sauces, tobacco, and tobacco smoke particulates.

Total N-nitroso compounds (NOC) and NOC precursors (NOCP) were determined in extracts of food and tobacco products. Following Walters' method, NOC were decomposed to NO with refluxing HBr/HCl/HOAc/EtOAc and NO was measured by chemiluminescence. NOC were determined after sulfamic acid treatment to destroy nitrite, and NOCP were determined after treatment with 110 mM nitrite and then sulfamic acid. Analysis without HBr gave results < or =20% of those with HBr. This NOC method was efficient for nitrosamines but not nitrosoureas. The standard nitrosation for determining NOCP gave high yields for readily nitrosated amines, including 1-deoxy-1-fructosylvaline, but not for simple amines, dipeptides, and alkylureas. Mean NOC and NOCP results were (respectively, in micromol/kg of product) 5.5 and 2700 for frankfurters, 0.5 and 660 for fresh meat, 5.8 and 5800 for salted, dried fish, and 660 and 2900 for chewing tobacco (all for aqueous extracts) and 220 and 20000 nmol/cigarette for MeCN extracts of cigarette smoke filter pads.     

Flavor and texture of banana chips dried by combinations of hot air, vacuum, and microwave processing

The behavior of 16 volatile compounds of banana during a combination of air-drying (AD) and vacuum microwave-drying (VMD) of banana chips was characterized. Samples were AD to remove 60, 70, 80, or 90% of moisture (wet basis) and then subjected to VMD to achieve a final moisture content of 3% (dry basis). Banana slices were also dehydrated using only AD, VMD, and freeze-drying (FD) for comparison. Samples that underwent more VMD had significantly lower levels of volatile compounds, which is attributed to the decreased formation of an impermeable solute layer on the surface of the chips. High values for water solubility and relative volatility of compounds correlated with losses during VMD; however, additional factors appear to influence the behavior of compounds during VMD processing. The optimal process of 90%AD/10%VMD yielded crisper banana chips with significantly higher volatile levels and sensory ratings than AD chips.     

Quantitative determination of coumestrol in dried alfalfa and alfalfa leaf protein concentrates containing chlorophyll.

An improved analytical method for the determination of coumestrol in dried alfalfa and leaf protein concentrates is described. In this method, chlorophyll is removed from an alcohol extract prior to the paper chromatographic-fluorometric measurement of coumestrol. Ninety-eight per cent of the coumestrol added to alfalfa leaf protein concentrates is recovered by this method. This improved method gives replicate values with lower standard deviations and coefficients of variation than the literature method.     

Emulsifying ability of porphyran prepared from dried nori, Porphyra yezoensis, a red alga

A suspension of low-quality dried nori processed from Porphyra yezoensis, a red alga, was autoclaved at 120 degrees C for 30 min, and from the supernatant, five preparations of porphyran of differing molecular masses and chemical compositions were obtained by preprecipitation with ethanol at stepwise-increasing concentrations of 50 and 67% followed by size-exclusion chromatography. The porphyran preparations exhibited a high emulsifying activity index and high emulsion stability over a wide range of pH and temperature and also in the presence of sodium chloride. An adequately high coefficient of correlation between the median diameter of oil droplets and their 3,6-anhydrogalactose content suggests that 3,6-anhydrogalactose could take part in emulsification with porphyran.