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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

Induction of favism-like symptoms in the rat: Effects of vicine and divicine in normal and buthionine sulphoximine-treated rats

Rats were fed an adequate or a deficient diet and offered water or buthionine sulphoximine (BSO) solution for 2 weeks, and then the same diets with vicine for another week in experiment 1. BSO in combination with the deficient diet caused a marked decrease in blood glutathione (GSH) and growth retardation but failed to show any effects resulting from supplementation with vicine. In experiment 2 the rats were given an adequate diet and BSO as before, and injected intravenously with divicine (DV). Here again, BSO depressed rat growth, and so did DV. Each of the insults also caused haematological changes, especially a fall in GSH, but the most severe changes appeared in the group treated with both BSO and DV. A decrease in haematocrit and increases in adrenal and spleen weight were also noted. In experiment 3 the rats were injected with different doses of DV, without pretreatment with BSO. The main effect was a drop in blood GSH and haematocrit, and an increase in adrenal and spleen weights, all of which were dose-related. Administration of the higher doses of DV resulted in a severe cyanosis followed by death within a relatively short period of time.     

Favism-like effects of divicine and isouramil in the rat: Acute and chronic effects on animal health,mortalities, blood parameters and ability to exchange respiratory gases

In vivo and in vitro experiments were carried out to establish the effects of divicine (DV) and isouramil (IU) on the rat when administered intra-peritoneally (IP). The LD₅₀for DV and IU was 148 and 183 mg kg^?1 body weight, respectively. There was a high negative correlation between the amount of DV added in vitro to blood and the blood glutathione (GSH) concentration, the hemoglobin absorbance ratio (576 nm/630 nm) and the erythrocyte sedimentation rate. The same relationships occurred in vivo, with mortalities also being highly correlated with these factors. Intravenously injected DV completely disappeared from the blood within a few minutes, indicating a very short half-life. The concentration of glutathione (GSH) in the red blood cells (RBC) was much higher and affected to a much greater degree in vivo by D V compared with GSH in the plasma. In the fourth experiment successive sub acute injections of DV and IU increased spleen weight and adrenal weights (IU only) and reduced kidney weights. Packed cell volume, hemoglobin concentration, leucocytes, lymphocytes, monocytes, and urea-N were also affected. Blood gas analyses in the fifth study showed that both DV and IU markedly reduced the partial pressure of oxygen in the blood, the per cent oxygen saturation, the pH and the bicarbonate concentration. This suggests that these compounds impaired the functional capacity of the RBC. The concentration of GSH was lower in RBC from young rats, and young rats, as compared with more mature rats, were considerably more sensitive to the toxic effects of DV. This work provides direct evidence, for the first time, on many of the adverse effects of these compounds.     

A modified procedure for the purification of vicine and convicine from fababeans (Vicia faba L.)

A procedure was developed for the simultaneous isolation of pure vicine [2,6-diamino-4,5-dihydroxypyrimidine 5-(β-D glucopyranoside)] and convicine [2,4,5-trihydroxy-6-aminopyrimidine 5-(β-D glucopyranoside)] from dehulled fababeans. The procedure involved the extraction of the glycosides with an ethanol-water (70:30, by vol.) solution, concentration of the extract to a minimal volume and adjustment of the pH of the aqueous extract to 7.2. Mixed vicine and convicine crystals, which were formed over a period of from 1 to 4 days, were harvested by centrifuging and washed with water and ethanol. Vicine was readily obtained from the mixed crystals by extraction at a low pH and recrystallised by adjustment of the pH to neutrality. Convicine, which is relatively insoluble in an acidic medium, was solubilised in an alkaline solution and recrystallised at a neutral pH.     

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     

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.     

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.     

Effect of particle size of pea (Pisum sativumL) flours on the digestion of their proteins in the digestive tract of broilers

Pea (Pisum sativum L) protein digestion in relation to particle size was studied in broilers. Birds were fed coarse pea (CP) or fine pea (FP) or nitrogen-free (NF) diets. Protein digestion was studied in gastro-intestinal contents of chicks by gel filtration and electrophoresis. Apparent ileal protein digestibility was lower for CP (70·2%) than FP (89·5%) diets. A decrease of protein digestibility appeared between ileum and excreta for FP diets. Endogenous proteins in ileal digesta represented important amounts. Estimation of excreta composition was about 24% of microflora in FP and CP. The molecular weight (MW) dis-tribution of nitrogen compounds of diets were characterised by high proportions of components with MW>15000. The relative amounts of very low MW (<500) components were in all cases lower for CP than for FP digesta. In intestine, proportions of high MW (>15000) components were higher for CP than for FP fed birds. Intestinal digesta of NF diet were characterised by higher MW than FP digesta. Similar electrophoresis patterns appeared for CP digesta and pea diet, whereas fine particles of CP and FP showed disappearance of protein bands. It is concluded that the CP diet leads to lower digestion of proteins than FP, but FP diets can lead to low MW proportion in intestine, which could be a limiting factor in pea protein digestion. © 1997 SCI.     

Supplementation of pig diet with algal fibre changes the chemical and physicochemical characteristics of digesta

Seaweed extracts, because of their physicochemical characteristics and potential nutritional value, could provide a new source of dietary fibre. This study investigated changes in seaweed fibres (physicochemical and fermentative properties) in different digestive sites and their effects on digesta (viscosity and hydration properties). Sixteen pigs were adapted to a test diet supplemented with 5% algal fibre (either Palmaria palmata (PP), a poorly viscous soluble xylan; or Eucheuma cottonii (EC), a partly insoluble carrageenan; or Laminaria digitata (LD), a highly viscous soluble alginate) or 5% cellulose (reference fibre). PP did not modify the characteristics of digesta and was fermented in the caecum (pH 6.1 ± 0.4; short‐chain fatty acids measured in digesta, 1409 ± 691 µmol g⁻¹ dry matter). EC and LD were mainly insoluble in the stomach, becoming soluble in the intestine; EC was slightly fermented in the colon, giving a low concentration of short‐chain fatty acids (303 ± 122 µmol g⁻¹ dry matter). Supplementation of the diet with alginate (LD) increased 3.5‐fold the ileal viscosity of digesta and their hydration capacity in the ileum and colon. Thus the physicochemical properties of pig digesta largely depend on the physicochemical properties of the ingested seaweed fibre, the pH and ionic conditions prevailing in the gut, and their fermentability. © 2000 Society of Chemical Industry     

Short communication: Effect of abomasal inorganic phosphorus infusion on phosphorus absorption in large intestine,milk production,and phosphorus excretion of dairy cattle

The objective of the study was to evaluate the effect of inorganic phosphorus (Pi) infusion on P absorption in large intestine, milk production, and phosphorus excretion. Four ruminally and ileally cannulated crossbred cows were used in a 4 × 4 Latin square with 21-d periods. Cows were fed a total mixed ration containing 0.21% P, providing 50% of the cows' P requirement. Cobalt-EDTA was used as marker to measure large intestine digesta flow. On d 13 to 21 of each period, each cow was infused daily with 0, 20.1, 40.2, or 60.3 g of Pi into the abomasum and total collection was conducted on d 18 to 21. Ileal samples were collected every 9 h on d 18 to 21. Feed, digesta, and fecal samples were analyzed for total P and Pi using the molybdovanadate yellow method and blue method, respectively. All data were analyzed using PROC GLIMMIX in SAS 9.3 (SAS Institute Inc., Cary, NC) using contrasts to evaluate linear, quadratic, and cubic effects of Pi infusion dose. Dry matter intake, apparent dry matter digestibility, milk yield, and milk total P were unaffected by Pi infusion. Ileal flow and fecal excretion of total P and Pi increased linearly with increasing infused Pi. In the large intestine, net absorption of TP and Pi was increased linearly with increasing infused Pi. The magnitude of absorption from the large intestine was greater than reflected in current models, raising questions that could be evaluated with longer infusion periods or dietary alteration.     

Digestion and absorption of lipids by sheep fed chopped and ground dried grass

Three wether sheep equipped with re-entrant cannulae at the proximal duodenum and terminal ileum were given dried cocksfoot in the chopped or ground form and the samples of duodenal and ileal digesta and faeces obtained for both diets were analysed for individual long-chain fatty acids. The total quantities of fatty acids entering the proximal duodenum (15.2 and 15.4 g/24 h on chopped and ground feed, respectively) were markedly higher than the amounts consumed (8.7 and 7.8 g/24 h, respectively), whilst losses within the small intestine were 13.9 and 14.2 g/24 h, respectively. None of these effects was found to be significantly influenced by the processing of the diet (P > 0.05). The quantity of C₁₈ unsaturated fatty acids in the two feeds comprised 78% of the total fatty acids but the level of these acids was considerably reduced (19%) in duodenal digesta and was not significantly influenced by diet (P > 0.05), whilst the levels of stearic acid (C_18:0) entering the small intestine were markedly increased on both diets.     

Nutritional and technological characteristics of new broad bean flaked products

The effects of the technological processes (soaking in water or alkaline solutions, drying, puree preparation) and the supplementation with maize flour on the nutritional value and on the organoleptic characteristics of broad bean (Vicia faba, L. major) flakes have been studied. Protein content was not affected by technological process. The addition of maize flour decreased the protein content of the final product depending on the amount of the maize flour added. Amino acid composition showed a decrease of tryptophan due to technological processing. Supplementation with maize flour improved the amino acid pattern and, except for tryptophan, the amount of essential amino acids in the flakes supplemented with 25% or more maize flour well compared with the provisional pattern by F.A.O. In vitro digestibility trials did not evidence significant changes due to technological processes or to integration of broad beans with maize flour. Broad bean toxic factors (vicine and convicine glycosides) were only slightly affected by the alkaline treatment of the flakes. Glycosides content decreased with the increasing supplementation with maize flour but the relationship was not linear. The organoleptic tests were positive for texture and taste, whereas the appearance of the products should be improved.