Purification,identification and quantification of an egg-weight-depressing factor (vicine) in fababeans (Vicia faba L.)

An egg-weight-depressing factor in fababeans was isolated in pure form from a white precipitate which had been shown in a previous study, to contain a concentrated form of this factor. The analytically pure compound was obtained in colourless, needle crystals. Its melting point was 242-244°C, it contained 17.4% nitrogen but did not react with ninhydrin. The spectra had adsorption maxima at 274, 276 and 269 nm in acidic, neutral, and alkaline solutions, respectively. These and other properties were identical to those of vicine [2,6-diamino-4,5-dihydroxy pyrimidine,5-(β-D-glucopyrano-side)] a compound known to be present in fababeans. Vicine and convicine [2,4,5-trihydroxy, 6-aminopyrimidine,5-(β-D-glucopyranoside)]; compounds having similar chemical and probably physiological activity, were resolved on a cation-exchange column and were detected by monitoring at 280 nm. Quantitative results obtained with the procedure was shown to be similar to those obtained with two other procedures. This method, however, was superior to other methods as it gave individual values for both vicine and convicine rather than a composite value; sample preparation was simple and the method could readily be adapted to automatic analysis.     

The estimation of vicine and convicine in fababeans (Vicia faba L.) and isolated fababean proteins

A specific method of analysis for each of the pyrimidine glucosides, vicine and convicine, is described. This involves extraction by ethanol, evaporation, separation on t.l.c. and quantitative ultraviolet spectrophotometry. Using this method mature dehulled fababeans were shown to contain on average 0.73% vicine and 0.30% convicine on a dry matter basis. No significant variation was found between varieties. The glycosides seem to be associated to the protein bodies in the raw bean, and are absent from the hull. The total content of vicine + convicine in isolated fababean proteins ranged from 0.07 to 2.0%. The highest content was found in a protein fraction produced by air-classification of dry milled fababeans. The lowest content was found in proteins isolated from a watery extract either by precipitation with acetic acid or by diafiltration. Protein isolates produced by direct ultrafiltration have contents of about 0.60%. The ratio vicine:convicine was found to be relatively constant in the raw beans (average 2.5), but varied from 1.4 to 3.0 in the protein isolates, thus indicating a different reaction of vicine and convicine to conditions during the processing of the fababean. It is the opinion of the authors that the etiology of favism is still controversial as far as vicine and convicine are concerned.     

The Distribution of Pyrimidinone Glucosides in Developing Seedlings of Vicia faba and Vicia narbonensis

The distribution of the pyrimidinone glucopyranosides vicine and convicine was investigated in resting seed and axenic developing seedlings up to 17 days of age in Vicia faba cv Troy, a low seed pyrimidinone glucopyranoside mutant (MTG5) derived from Troy, and a related species V narbonensis. The lower amounts of total vicine and convicine in resting seed of MTG5 were due primarily to a reduction in cotyledon pyrimidinone glucopyranosides, with the amount in the embryo axis much less affected. Distribution within developing seedlings revealed consistent patterns. A pulse of vicine appeared in both species in stems and roots at day 4 to day 7, then declined. Amounts of convicine rose initially then declined in seedling stems of both species, whereas in roots the rise in convicine continued throughout the period. The rise in convicine in the roots of V narbonensis could not have been accounted for by conversion from reserves of vicine. The low pyrimidinone glucopyranoside mutant MTG5 had less vicine initially in stems and roots than both Troy and V. narbonensis but by 17 days this had risen close to the levels of Troy and exceeded those of V narbonensis. By flowering, amounts of vicine and convicine in glass-house grown plants were similar in MTG5 and Troy. The levels of both compounds in the roots of MTG5 exceeded those of the sown seed. Consequently, both V faba and V narbonensis have the capacity to synthesise vicine and convicine in young plants. A mutation in a gene controlling levels of pyrimidinone glucopyranosides in the developing seed of V faba does not suppress the biosynthetic ability of young plants.     

Development of a simple procedure for the complete extraction of vicine and convicine from fababeans (Vicia faba? L.)

Egyptian foods prepared from fababeans contain variable and relatively high concentrations of vicine. Vicine and convicine can be readily extracted from whole or dehulled fababeans. The rate and completeness of extraction depends upon size of beans, the presence or absence of testa, the pH, temperature and volume of the aqueous extracting solution, the duration of the extraction procedure and the frequency with which the extracting solution is changed. In a typical experiment vicine and convicine were almost completely removed from whole or dehulled fababeans when they were extracted at 20 or 40°C with 1% glacial acetic acid (0.17M) or water over a 72 h period with changes in the extracting media every 24 h. This appears to be the most practical procedure for extraction. The low pH of the acid solution prevents bacterial fermentation that occurs when water is used. Water at temperatures greater than 70°C is a highly effective extracting agent. Extraction of fababeans with acetic acid does not markedly affect the concentration of protein (N × 5.7), fat or fibre. Ash content, however, was reduced by a factor of 2 and the total dry matter content of the extracted beans was reduced by 16–17% in whole beans and 20–21% in dehulled beans.     

The concentration of vicine and convicine in Vicia faba and some related species and their distribution within mature seeds

Mature seeds from a range of species from the Vicieae tribe were analysed for vicine and convicine content. Quantifiable amounts of vicine were found in all the Vicia species analysed but neither glucopyranoside was detected in Pisum or Lathyrus. The concentrations of both vicine and convicine were similar for both Vicia faba and V bithynica but in all the other Vicia species glucopyranoside concentrations were significantly lower. Seeds from three faba bean lines and one each from V narbonensis and V bithynica were dissected into their constituent seed parts and these parts analysed. Only trace quantities were found in the seed coat but concentrations of vicine ranging from 39 g kg^?1 to 81 g kg^?1 were detected in the radicle samples. The implications of these results in relation to the role of these compounds in the plant are discussed.     

Removal of favism-inducing factors vicine and convicine and the associated effects on the protein content and digestibility of fababeans (Vicia faba L)†

Ingestion of fababeans is associated with precipitation of the haemolytic disease favism in certain glucose-6-phosphate dehydrogenase-deficient humans. Hence their incorporation into weaning foods has not yet been practised. Few, if any, attempts to detoxify fababeans have so far been made. The present investigation has evaluated the effects of a series of treatments designed for vicine and convicine extraction and hydrolysis followed by oxidation of their pyrimidine moieties (divicine and isouramil). The effects of seed germination and oxidative treatment by hydrogen peroxide have also been examined. Extracts of treated samples were assayed for residual vicine and convicine. Toxicity was monitored by the changes in vitro of reduced glutathione (GSH) of red blood cells (RBCs) from Sprague Dawley rats deliberately made deficient in glutathione reductase activity to mimic favism-susceptible human RBCs. Treatments of whole cotyledons resulted in recovery of 59–93% vicine and 50–70% convicine originally present in the seeds. Treatments of fababean powders, however, were capable of lowering the vicine content by 94–100% and convicine content by 100%. Germinated seeds showed a drop in vicine content of 86% and their hydrogen peroxide treatment 91–93%. Convicine was totally absent in germinated and oxidised seeds. The results of toxicity assays were concomitant with vicine and convicine analyses. Protein content of original fababean powders was well recovered (up to 94.00%) and its digestibility was almost complete (99.34%). © 1999 Society of Chemical Industry     

Turnover and hydrolysis of vicine and convicine in avian tissues and digesta

The objectives of this study were to establish factors influencing the absorption, excretion and hydrolysis of vicine and convicine in chicks. Blood vicine, following the oral administration of a single dosage of vicine into the crop of young chicks, reached maximum concentrations within 3 h. It was nearly completely removed from the blood within 12 h and had a half-life of approximately 4.5 h. The accumulation and excretion patterns of vicine in the liver and kidney were similar to those of the blood except that the concentrations were much higher in these tissues, particularly the kidney. Bile also contained a very high concentration of vicine which tended to accumulate following the decline in other tissues. These results together with the appearance of vicine in the urine of colostomised birds suggest that vicine is excreted in the urine and bile. Convicine in contrast to vicine was not absorbed by the chick. In-vitro studies were carried out with tissue and digesta homogenates from the chick in order to establish the site(s) at which vicine and convicine were hydrolysed to their aglycone forms. The results demonstrated that neither vicine nor convicine were hydrolysed in the presence of liver, kidney, intestinal wall or caecal wall homogenates, digesta from the large intestine or by enzymes present in whole or ground fababeans. They were, however, slowly hydrolysed in the presence of 0.1N HCl at 37°C and very rapidly hydrolysed by digesta from the caeca. Antibiotic additions to the diets markedly reduced the in-vitro rate of hydrolysis of these compounds. The latter results suggest that vicine and convicine are hydrolysed by microorganisms in the caeca of the chick but are not hydrolysed by the micro-organisms in the gastrointestinal tract, by endogenous tissue enzymes or by enzymes present in fababeans and only minimally hydrolysed by the low pH of the stomach.     

Extraction of favism‐inducing agents from whole seeds of faba bean (Vicia faba L var major)

The objective of the present study was to establish a suitable procedure by which favism‐inducing glycosides vicine and convicine could be completely removed from whole seeds of faba beans without distorting their shape or reducing their nutritive value. Detoxification procedures carried out were extraction by water or, 10 g l^?1 acid solutions (acetic, citric and phosphoric acid) using stepwise or continuous soaking procedures. Three cultivars of the beans were used in the study. In the stepwise soaking procedures, the seeds were soaked in either water or acid solution at 40 °C for different periods while in the continuous flow techniques, the solutions were constantly passed through a seed reservoir under different time–temperature–flow rate conditions. Extraction of vicine and convicine increased with increasing temperature and period of soaking and/or continuous flow of soaking solutions. Neither stepwise soaking nor autoclaving could effectively remove the favism factors from the whole seeds. The best results were obtained with continuous flow soaking in tap water, giving vicine‐ and convicine‐free seeds. The protein contents of treated seeds were only slightly affected and their physical structure remained intact. Thus, it was concluded that the latter procedure was the method of choice for detoxification of dry whole seeds. Copyright © 2005 Society of Chemical Industry     

Metabolism of vicine and convicine in rat tissues: absorption and excretion patterns and sites of hydrolysis

The objectives of this study were to determine whether dietary vicine and convicine were absorbed by rat tissue, and to determine their excretion patterns and/or sites of degradation. Orally administered vicine and convicine were excreted in relatively low amounts via the kidney and faeces. However, no vicine or convicine was detected in the blood, liver, kidney or muscle tissue of rats which had been fed these compounds. In-vitro studies demonstrated that vicine and convicine were not hydrolysed in liver, kidney, muscle, caecal wall or intestinal wall homogenates. In contrast, digesta samples from the large intestine and caeca were able to rapidly hydrolyse these compounds, with the concomitant formation of new compounds. Digesta from the stomach and small intestine promoted the slow hydrolysis of these compounds, as did fresh faecal samples. These results would suggest that vicine and convicine are absorbed by the rat in only limited amounts, are not hydrolysed by rat tissues, and are rapidly cleared from tissue via the kidney. The bulk of the dietary vicine and convicine are hydrolysed in the large intestine and caecum.     

Hydrolysis of the toxic constituents (vicine and convicine) in fababean (Vicia faba L.) food preparations following treatment with β-glucosidase

Several experiments were carried out to determine whether the concentrations of vicine and convicine in fababean food preparations were reduced following heat and β-glucosidase treatment. Heating fababeans to relatively high temperatures was not effective in reducing the concentration of these glycosides. Treatment of cooked fababeans (fababean paste) with ground raw almond powder, which is a rich source of β-glucosidase (EC 3.2.1.21) activity, resulted in the hydrolysis of vicine and convicine. The degree of hydrolysis ranged from partial to complete and was dependent on the duration of the treatment period (1 to 24 h), the temperature at which treatment was carried out (1 to 30°C), the pH (6.4 to 4.6) as adjusted by the addition of lemon juice and the amount of ground almonds added to the fababean paste. For example, 88 to 89% of the glycosides were hydrolysed when 1 g of fababean paste was mixed with 0.1 g of almond powder and 0.1 ml of lemon juice and incubated at 30°C for 3 h. This study demonstrated that the concentration of vicine and convicine and their corresponding aglycones can be greatly reduced in fababean food preparations by enzymic hydrolysis.     

Effects of intraperitoneally injected vicine and convicine on the rat: Induction of favism-like signs

Rats were injected intraperitoneally with single or several successive doses of vicine or convicine. Single injections of these compounds at different concentrations caused increased respiration rates, generalised cyanosis, abdominal convolutions and, after several hours, death which appeared to be caused by asphyxiation. The tissues of the dead animals were engorged with dark brown blood and the large intestine and caecum contained entrapped gases and watery digesta and faecal matter. A second study demonstrated that injected vicine and convicine were cleared from the intraperitoneal cavity over several hours via the kidney and large intestine and that they were cleaved in the digesta of the large intestine and colon to divicine and isouramil. In a third study, rats were injected daily for 10 days with vicine or convicine. There were increases in spleen weight and blood monocytes and neutrophil counts and decreases in liver weight, blood glutathione and glucose concentrations, and lymphocyte counts. Blood from rats pretreated in vivo with convicine was shown to have an altered ultraviolet absorbance pattern. A similar pattern developed in vitro only in the presence of the aglycones of vicine (divicine) or convicine (isouramil) but not in the presence of the compounds themselves. The results of these studies demonstrate that vicine and convicine when injected intraperitoneally into the rat are converted to their aglycones which cause signs similar in many respects to those observed in the human metabolic disease, favism.     

Effect of genotype and environment on vicine and convicine levels in fababeans (Vicia faba minor)

The concentrations of vicine and convicine (1.94 and 0.83%, dry basis, respectively) in fababeans were highest in young seeds and decreased rapidly with maturity. By 21-28 August 1978 the concentrations had levelled off and the rank order of four cultivars, which differed widely in days required to mature, remained unchanged for the rest of the growing season. Analysis of variance on 36 fababean cultivars grown at different locations and years in Saskatchewan showed that genotype and environment had highly significant (P=0.01) effects on the glucoside level of fababean samples. Although lower in concentration (mean=0.31%), convicine showed greater variability among cultivars than vicine (mean = 0.66%). The glucoside contents of 242 diverse cultivars grown at Saskatoon ranged from 0.44 to 0.82% for vicine and 0.13 to 0.64% for convicine with the lowest cultivar containing 0.62% total glucoside. Individual seeds of a cultivar differed greatly in glucoside content and could become another source of variability for plant breeding and selection of low-glucoside cultivars. Hulls were relatively free of glucosides but, despite variations in hull content, it was unnecessary to dehull samples before analysis.     

Nutritional and functional properties of Vicia faba protein isolates and related fractions

The goal of this research was the characterisation of Vicia faba (broadbean) protein isolates and related fractions in order to determine whether this grain legume could be used for production of high quality protein products and other fractions rich in functional components. Alkaline extraction of the defatted seed flour, followed by precipitation at the isoelectric pH, yielded a 92% protein isolate with a high oil absorption capacity. The contents of the favism-inducing glycosides, vicine and convicine, in the isolate were reduced by more than 99% as compared to the original flour, although the amino acid composition was similar to that of the flour. Some of the by-products of protein isolate production may also be of interest from a nutritional and functional point of view. Thus, the oil resulting from hexane extraction of the flour is rich in unsaturated fatty acids, and polyphenols (resulting from extraction of the defatted flour with acetone) showed a high ABTS radical-scavenging activity. In addition, the solid residue (resulting from protein solubilisation) was high in fibre and showed good water absorption. These results show good nutritional and functional properties in V. faba protein isolates and related fractions, which may favour the revalorisation of this traditional bean crop.     

Content and distribution of vicine,convicine and <Emphasis Type="SmallCaps">l</Emphasis>-DOPA during germination and seedling growth of two <Emphasis Type="Italic">Vicia faba</Emphasis> L. varieties

This work examined the content and distribution of the pyrimidine glucosides vicine and convicine, and the nonproteic amino acid l-DOPA in cotyledons and embryo axes along the germination and seedling growth of Vicia faba L. vars. Alameda and Brocal. The behaviour of these compounds was similar in both varieties. Vicine and convicine, implicated in favism disease, slowly declined in cotyledons. In embryo axis, vicine levels were sharply reduced and convicine amount was slightly increased as assay progressed. Total pyrimidine glucosides remained unchanged in the whole plant during the study. l-DOPA only appeared in the embryo axis of V. faba seeds and the highest content was observed in Brocal variety at 6 DAI (days after imbibition). The information provided in this study could be valuable for a possible role of embryo axis, rich in l-DOPA, for the treatment of Parkinson’s disease and also as a choice ingredient for functional foods or as nutraceutical.     

Recovery of proteins from beef bone and the functionality of these proteins in sausage batters

Four solutions [4% sodium chloride (control), 4% sodium chloride with 0.3% sodium tripolyphosphate (STP), 0.3% tetrasodium pyrophosphate (TTP) or 0.05 M sodium hydroxide (NaOH)] were used to extract proteins from beef bones. Three bone solution ratios (1:1, 1:4 or 1:10), three bone types [vertebra (lumbar), rib (4-7) and leg (femur)] and two methods of protein recovery from the extraction slurries [dialysis against 0.03 M potassium phosphate buffer (pH 5.3) and acid precipitation] were evaluated. Solutions containing phosphates or NaOH were more effective in extracting protein than sodium chloride alone. Total protein recovery was highest from vertebra bones while extraction of proteins from leg bones resulted in the lowest recovery. A solution to bone ratio of 1 to 10 recovered more total protein from vertebra or rib bones than leg bones. Dialysis recovered more total protein from extraction solutions when the protein concentration was low. Acid precipitation, however, worked best if the protein concentration in the extraction solution was high. Extraction procedures resulted in some myosin degradation. Proteins extracted from beef bone materials and recovered by dialysis, performed equally as well as other commercially available proteins when added to a finely comminuted sausage product.     

A one-pot ultrasound-assisted water extraction/cyclodextrin encapsulation of resveratrol from Polygonum cuspidatum

The ultrasound-assisted extraction (UAE) of resveratrol and other polyphenols from Polygonum cuspidatum has been carried out with the aim of developing an efficient and eco-friendly extraction process. The finely milled roots were sonicated (titanium horn, 19.5 kHz) in methanol, in different cyclodextrin water solutions [β-cyclodextrin (β-CD) or hydroxypropyl β-CD (HPβ-CD)] and also in pure water. UAE dramatically increased the yields and cut down extraction times compared to conventional extraction under stirring. Outstanding results have been achieved with the β-CD solution (1.5% w/w); in fact HPLC analysis showed that the selective inclusion properties of CDs toward phenolic stilbenes gave a much cleaner analytical extract profile. This green method gave 7.51 mg of total resveratrol (free + cleaved polydatin glucoside) per gram of dry plant. Thanks to polyphenol encapsulation within CDs, this extract showed excellent water dispersibility, higher stability and an antioxidant power which is comparable to that of the MeOH extract (DPPH, ORAC_FL). These important features should pave the road for its application in food supplements or phytochemical preparations.     

富含黄酮和生物碱的桑叶提取物的提取工艺研究

目的：研究桑叶黄酮和生物碱的同步高效提取工艺,通过正交试验获得最佳提取方案。方法：在单因素试验基础上用正交试验进行工艺参数的优化,获得最佳提取方案。结果：桑叶黄酮和生物碱的最佳同步提取工艺为液料比10:1(V/m)、乙醇浓度60%(pH2)、提取时间2h、提取温度80℃、提取2 次。结论：用该法对桑叶黄酮和生物碱进行共同提取,提取率较高,可以获得二者含量较高的提取物。     

Selective removal of 226Ra2+ from gas-field-produced waters

The 226Ra2+ selectivity of both the self-assembled (iso)-guanosine-based systems and ionizable thiacalix[4]crown dicarboxylic acids was determined in gas-field-produced water and a metal ion-containing model solution (simulant). Seven gas-field-produced water samples have been analyzed. From a sample (K5D) with average metal ion concentrations ([metal(tot)] = 0.14 M), thiacalix[4]crown-5 dicarboxylic acid (10(-4) M) extracts 60% of the 226Ra2+ content. Extractions performed with the model solution ((M)K5D) indicate that in K5D there is significant competition in 226Ra2+ extraction due to the organic constituents of K5D, in particular with self-assembled extractants guanosine and isoguanosine. Nevertheless, all four extractants extract 226Ra2+ both from the produced water K5D and the model solution (M)K5D, even with a 100-fold excess of [metal(tot)] to [extractant]. The extracted 226Ra2+ cations could effectively be stripped from the extractants bywashing with pH 2 water. The results obtained with the extractants used, especially thiacalix[4]crown-5 dicarboxylic acid 3, clearly demonstrate the way to selectively remove Ra2+ from gas-field-produced waters.     

酶解辅助反胶束法萃取小麦胚芽蛋白的研究

采用由AOT(琥珀酸二(2-乙基己基)酯磺酸钠)-异辛烷-KCl缓冲液组成的反胶束体系,联合酶解技术提取小麦胚芽蛋白。探索了酶与底物浓度比、时间、缓冲液pH、温度四个因素对蛋白质前萃率的影响。结果表明:酶解辅助反胶束萃取蛋白是可行的,可以显著提高前萃率,最佳酶与底物浓度比为0.23AU/gPro、时间8h、缓冲液pH8.10、温度60℃。     

杨梅渣红色素酸水提取和精制工艺

以新鲜杨梅渣为原料,利用正交试验法确定酸水提取杨梅渣红色素最佳工艺条件为料液比1:12(g/mL)、提取温度60℃、提提时间4h、pH1.5盐酸。采用离子交换树脂法对色素进行精制,在供试的5种树脂中D002的精制效果最好;通过静态吸附、解吸实验以及动态吸附实验,确定D002离子交换树脂精制杨梅渣红色素的工艺条件为杨梅渣红色素吸光度0.5～1、色素pH2～3、25℃左右、吸附时间1.5h、上样流速3BV/h。