Identification and quantification of key phytochemicals in peas - Linking compounds with sensory attributes.

Pea protein isolates contain high-quality plant protein. However, they have sensory drawbacks, notably bitterness and astringency, that have limited their use in commercial foods. This study's aim was thus to identify the main phytochemicals in pea-based samples and to examine associations with sensory attributes. The phytochemical profiles of pea flour, pea protein isolates, and pea protein isolate fractions were characterized via UHPLC-DAD-MS. A total of 48 phytochemicals have been revealed: 6 phenolic acids, 5 flavonoids, and 1 saponin were identified and quantified, while another 9 phenolic acids, 10 flavonoids, and 6 saponins were tentatively identified. The impacts of protein extraction and fractionation were studied. These processes appear to have caused some compound degradation. It was found that 29 compounds were correlated with perceived bitterness and/or astringency. Therefore, these results show that certain phytochemicals can lead to negative sensory attributes in pea-protein-based products.

Potent aroma compounds of two red wine vinegars.

Gas chromatography olfactometry (GCO) was used to determine key aroma compounds of two red wine vinegars. Sensory analysis was performed to choose the best neutralization agent of acetic acid (NaOH or MgO) and to test representativeness of four extracts obtained by different methods (dichloromethane extraction, XAD-2, mixture of XAD-2 and XAD-7, and Extrelut resins extraction). Neutralization with NaOH followed by dichloromethane extraction was selected to extract volatile compounds of vinegars. Key odorant compounds were determined by GCO based on detection frequency with 13 people. In the two red wine vinegars, 13 odors were perceived by at least 70% of the panelists, and 8 compounds among the 13 were identified: acetic acid, 3-methylbutyric acid, 2-phenyl-1-ethanol, 2, 3-butanedione, butyric acid, 2-methylbutyric acid, mixture of 2- and 3-methyl-1-butanol, and two newly identified compounds in vinegar, 3-hydroxy-2-pentanone and 3-(methylthio)-1-propanal. Quantification of all the volatile compounds was performed by GC-FID, and 10 other compounds were identified for the first time in wine vinegar.

Lymphatic absorption of phytosterol oxides in rats.

Two of the main classes of oxyphytosterols (7-keto and epoxides) were synthesized from sitosterol and campesterol and given to mesenteric duct-cannulated adult male rats. Lymph was collected during 24 h and was analyzed for oxysterols. The results showed that the lymphatic recovery of the phytosterol oxides was low: 4.7% of the given dose for epoxy derivatives and 1.5% for 7-keto compounds. The campesterol oxides presented a better absorption than the sitosterol oxides. During the process of absorption, the epoxyphytostanols were also partly transformed in campestanetriol and stigmastanetriol.

Geometry of conjugated double bonds of CLA isomers in a commercial mixture and in their hepatic 20:4 metabolites.

Rats were fed a fat-free diet for 2 wk. After this period, while maintaining the animals on the same diet, the rats were given intragastrically 180 mg per day of a mixture of conjugated linoleic acids (CLA) as triacylglycerols. Gas chromatography mass spectrometry (GC-MS) analyses of this mixture, as well as hydrazine reduction and GC-MS and GC-Fourier transform infrared analyses of the resulting monoenes, revealed the presence of two major isomers, the 9c,11t- and the 10t,12c-18:2 accompanied by smaller amounts of the 8t, 10c and the 11c, 13t-18:2 isomers. Minor quantities of cis,cis and trans,trans conjugated isomers also were detected. The total fatty acid methyl esters from the liver lipids were fractionated by reversed-phase high-performance liquid chromatography, and the fraction containing the 20:4 isomers was further fractionated by silver nitrate thin-layer chromatography. A band containing two 20:4 conjugated isomers was submitted to hydrazine reduction and the resulting monoenes analyzed by GC-MS as dimethyl-oxazoline derivatives. The two conjugated isomers were tentatively identified as 5c,8c, 11c, 13t-20:4 and 5c,8c,12t,14c-20:4. These could be formed by desaturation and elongation of the 9c,11t- and 10t,12c-18:2 present in the commercial CLA mixture.

Incorporation and metabolism of trans 20:5 in endothelial cells. Effect on prostacyclin synthesis.

To study the ability of long-chain trans fatty acids (FA) to be incorporated and metabolized into endothelial cells, bovine aortic endothelial cells were incubated with medium enriched eicosapentaenoic acid (EPA) bound to albumin (M2) or one of its geometrical isomers: 20:5 5c,8c,11t,14c,17c (M3), 20:5 5c,8c,11c,14c,17t (M4), or 20:5 5c,8c,11t,14c,17t (M5). After 48 h of incubation, supernatant and cells were harvested and their lipids were analyzed, including prostacyclin synthesis. EPA and 22:5n-3 of endothelial cells incubated with M2 were, respectively, three and two times higher than in control cells (incubated in M1, without any fatty acid added), whereas 22:6n-3 increased only in the supernatant, suggesting its release after biosynthesis. However, 18:2n-6 and 22:4n-6 decreased (about 30%). Trans 20:5 isomers represented 4.7, 3.9, and 5.2% of total phospholipid FA in endothelial cells incubated with M3, M4, and M5, respectively. They were elongated into trans 22:5 and trans 24:5, as revealed by gas chromatography-mass spectrometry and gas chromatography-Fourier transform infrared analysis. In cells incubated with M2, M3, M4, and M5, prostacyclin synthesis was inhibited by 49.0, 62.5, 60.5, and 72.0%, respectively. This effect may be due to less available arachidonic acid in the cells and to a competition between EPA isomers and AA at the level of cyclooxygenase pathway, as it was demonstrated that 20:5 delta17t was metabolized by this enzyme.

In vivo metabolism of diallyl disulphide in the rat: identification of two new metabolites.

1. Diallyl disulphide (DADS), a compound formed from the organosulphur compounds present in garlic, is known for its anticarcinogenic effects in animal models. 2. The aim was to identify and analyse the metabolites produced in vivo after a single oral administration of 200 mg kg(-1) DADS to rats. The organic sulphur metabolites present in the stomach, liver, plasma and urine were measured by gas chromatography coupled with mass spectrometry over 15 days. 3. Data indicate that DADS is absorbed and transformed into allyl mercaptan, allyl methyl sulphide, allyl methyl sulphoxide (AMSO) and allyl methyl sulphone (AMSO(2)), which are detected throughout the excretion period. Overall, the highest amounts of metabolites were measured 48-72h after the DADS administration. AMSO(2) is the most abundant and persistent of these compounds. The levels of all the sulphur compounds rapidly decline within the first week after administration and disappear during the second week. Only AMSO and AMSO(2) are significantly excreted in urine. 4. These potential metabolites are thought to be active in the target tissues. Our data warrant further studies to check this hypothesis.

Analysis of metabolic pathways by the growth of cells in the presence of organic solvents.

A new approach to the analysis of metabolic pathways involving poorly water-soluble intermediates is proposed. It relies upon the ability of the hydrophobic intermediates formed by a sequence of intracellular reactions to cross the membrane(s) and partition between aqueous and organic phases, when cells are incubated in the presence of a nonpolar and nontoxic organic solvent. As a result of this thermodynamically driven efflux of the formed intermediates from the cell, they accumulate in the organic medium in sufficient quantities for GC-MS analysis and identification. This enables direct determination of the sequence of chemical reactions involved with no requirement for the isolation of each individual metabolite from a cell-free extract. The feasibility of the proposed methodology has been demonstrated by the elucidation of the biosynthesis of (R)-gamma-decalactone from (R)-ricinoleic acid catalyzed by the yeast Sporidiobolus ruinenii grown in the presence of decane. The corresponding 4-hydroxy-acid intermediates, formed in the course of beta-oxidation of (R)-ricinoleic acid, were simultaneously observed in a single experiment on the same chromatogram. Potential applications of this proposed methodology are briefly discussed.

Novel scheme for biosynthesis of aryl metabolites from L-phenylalanine in the fungus Bjerkandera adusta.

Aryl metabolite biosynthesis was studied in the white rot fungus Bjerkandera adusta cultivated in a liquid medium supplemented with L-phenylalanine. Aromatic compounds were analyzed by gas chromatography-mass spectrometry following addition of labelled precursors ((14)C- and (13)C-labelled L-phenylalanine), which did not interfere with fungal metabolism. The major aromatic compounds identified were benzyl alcohol, benzaldehyde (bitter almond aroma), and benzoic acid. Hydroxy- and methoxybenzylic compounds (alcohols, aldehydes, and acids) were also found in fungal cultures. Intracellular enzymatic activities (phenylalanine ammonia lyase, aryl-alcohol oxidase, aryl-alcohol dehydrogenase, aryl-aldehyde dehydrogenase, lignin peroxidase) and extracellular enzymatic activities (aryl-alcohol oxidase, lignin peroxidase), as well as aromatic compounds, were detected in B. adusta cultures. Metabolite formation required de novo protein biosynthesis. Our results show that L-phenylalanine was deaminated to trans-cinnamic acid by a phenylalanine ammonia lyase and trans-cinnamic acid was in turn converted to aromatic acids (phenylpyruvic, phenylacetic, mandelic, and benzoylformic acids); benzaldehyde was a metabolic intermediate. These acids were transformed into benzaldehyde, benzyl alcohol, and benzoic acid. Our findings support the hypothesis that all of these compounds are intermediates in the biosynthetic pathway from L-phenylalanine to aryl metabolites. Additionally, trans-cinnamic acid can also be transformed via beta-oxidation to benzoic acid. This was confirmed by the presence of acetophenone as a beta-oxidation degradation intermediate. To our knowledge, this is the first time that a beta-oxidation sequence leading to benzoic acid synthesis has been found in a white rot fungus. A novel metabolic scheme for biosynthesis of aryl metabolites from L-phenylalanine is proposed.

Incorporation and metabolism of dietary trans isomers of linolenic acid alter the fatty acid profile of rat tissues.

To study the influence on lipid metabolism and platelet aggregation of the fatty acid isomerization that occurs during heat treatment, weanling rats were fed for 8 wk a diet enriched with 5% isomerized (experimental group) or normal (control group) canola oil. Geometrical isomers of alpha-linolenic acid representing 0.2 g/100 g of the experimental diet were incorporated into liver, platelets, aorta and heart, at the expense of their cis homologue and of 18:2(n-6). The major isomer, 9c,12c,15t-18:3, was also metabolized to 5c,8c,11c,14c,17t-20:5 and to an unknown compound, found in liver, platelets and aorta, which has been identified tentatively as 7c, 10c,13c,16c,19t-22:5. The greater 20:4(n-6)/18:2(n-6) ratio in the liver, platelets and heart of the experimental group than the control group indicated an enhancement of desaturation activities. This induced a higher content of long-chain (n-6) fatty acids in the experimental group. Platelet aggregation tended to be slightly higher (P: = 0.065) in the experimental group. We conclude that 0.2 g of trans isomers of alpha-linolenic acid per 100 g of diet was sufficient to be incorporated and metabolized, thus altering the fatty acid profile of rat tissues.

(Z)-dodec-3-en-1-ol, a novel termite trail pheromone identified after solid phase microextraction from Macrotermes annandalei.

(Z)-dodec-3-en-1-ol was isolated and identified by GC-MS as the major component of the trail-following pheromone from whole body and sternal gland extracts of workers of the fungus-growing termite, Macrotermes annandalei (Silvestri) (Termitidae, Macrotermitinae). For the first time, this trail pheromone was also identified by using solid phase microextraction from the surface of the secretory sternal gland of workers. Bioassays showed that synthetic dodecenol induced both orientation and recruitment behavioral effects. The activity threshold of (Z)-dodec-3-en-1-ol in eliciting trail-following is similar to that of (3Z,6Z,8E)-dodeca-3,6,8-trien-1-ol in the Rhinotermitidae, but amounts of dodecenol secreted are 100 times higher than those of dodecatrienol. There is about 1 ng of (Z)-dodec-3-en-1-ol per worker. Artificial trails made of synthetic dodecenol are able to compete with natural trails in the field. The activity duration of synthetic (Z)-dodec-3-en-1-ol trails is shorter than that of trails made from whole sternal secretion of workers. Observations showed that (Z)-dodec-3-en-1-ol is probably the only major component of the trail-following pheromone of M. annandalei and that it could be associated with other compounds in a pheromonal blend providing specificity and/or stability to trails.