The effect of short-term storage temperature on the key headspace volatile compounds observed in Canadian faba bean flour.

The odour emitted from the high-tannin fab bean flour (Vicia faba var. minor), was characterized by headspace solid-phase microextraction/gas chromatography-mass spectrometry (HS-SPME/GC-MS). The relative odour activity value (ROAV) was used to monitor the changes in key volatile compounds in the flour during short-term storage at different temperature conditions. The key flavour compounds of freshly milled flour included hexanal, octanal, nonanal, decanal, 3-methylbutanal, phenyl acetaldehyde, (E)-2-nonenal, 1-hexanol, phenyl ethyl alcohol, 1-octen-3-ol, ß-linalool, acetic acid, octanoic acid, and 3-methylbutyric acid; these are oxidative degradation products of unsaturated fatty acids and amino acids. Despite the low lipid content of faba beans, the abundances of aldehydes arising during room temperature storage greatly contributed to the flavour of the flour due to their very low odour thresholds. Two of the key volatiles responsible for beany flavour in flour (hexanal, nonanal) increased greatly after 2 weeks of storage at room temperature or under refrigerated conditions. These volatile oxidation products may arise as a result of enzymatic activity on unsaturated fatty acids, and was seen to be arrested by freezing the flour.

Sprouting improves the flavour quality of faba bean flours.

Flours were made from the sprouted seeds of the low- and high-tannin faba bean cultivars Fabelle, FB9-4, Snowbird, and Snowdrop. Headspace measurements on sprouted flours found the most favourable aroma profiles following 48 h sprouting and 24 h drying at 60 °C. Lipoxygenase activity, and the tannin, protein, and moisture contents were determined for unsprouted and sprouted faba bean flours. Lipoxygenase activity was higher in sprouted seeds before drying. Protein content increased after sprouting, whereas the tannin content decreased, especially for high-tannin varieties. Key volatile flavour compounds of faba bean flours included pentanal, hexanal, heptanal, octanal, nonanal, decanal, 1-hexanol, 1-octen-3-ol, 3-methylbutanal, phenyl acetaldehyde, 3-methylbutyric acid, d-limonene, ß-linalool, menthol, and estragole; these include oxidative degradation products of oleic, linoleic, and some amino acids. An overall flavour improvement was achieved after germination, as indicated by a decrease in bitter compounds (tannins) and beany flavours (hexanal, nonanal, 2-heptanone, and 2-pentylfuran).

Characterisation of the volatile flavour compounds in low and high tannin faba beans (Vicia faba var. minor) grown in Alberta, Canada.

The volatile flavour profiles of Canadian-grown faba beans (Vicia faba var. minor) were evaluated by headspace solid-phase microextraction (HS-SPME) coupled to gas chromatography-mass spectrometry (GC-MS). Two low- and high-tannin varieties were chosen, each in the form of dehulled and whole seed flours (DLT, DHT, WLT, and WHT, respectively). Pre-incubation time, fibre-extraction time, extraction temperature, and sample amount were evaluated during method optimization. The volatiles identified were classified into nine groups: aromatic hydrocarbons, aldehydes, alkanes, alkenes, alcohols, ketones, organic acids, esters, and others. Significant differences between dehulled and whole samples were found. Volatiles derived from amino acids were consistently observed in the volatile profile of all types. Despite the low lipid content of faba bean, significant amounts of volatiles normally associated with unsaturated fatty acids were present. HS-SPME/GC-MS proved to be a rapid, effective, and reproducible method (typical RSD < 5%) suitable for routine evaluation of faba bean volatile flavours.

Determination of organophosphorus and carbamate insecticides in fresh fruits and vegetables by high-performance thin-layer chromatography-multienzyme inhibition assay.

HPTLC-enzyme inhibition assay was applied to different fruit and vegetable samples after individual spiking with organophosphate and carbamate pesticides at their maximum residue limits documented by the European Commission. Samples were extracted according to the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) method, including cleanup by primary secondary amine sorbent. Additional cleanup was performed on the HPTLC plate by a prechromatographic step to separate most coextracted matrix compounds from 20 different pesticides under study. With both rabbit liver esterase and cutinase from Fusarium solani pisi as enzyme sources, mean recoveries from apples, cucumbers, grapes, nectarines, plums, tomatoes, and lemons were in the ranges 86-109, 95-129, 96-114, and 90-111% for chlorpyrifos, paraoxon, parathion, and pirimicarb, respectively, with a mean RSD of 8.5% for all samples.

Effect of bromine oxidation on high-performance thin-layer chromatography multi-enzyme inhibition assay detection of organophosphates and carbamate insecticides.

Following high-performance thin-layer chromatography, thiophosphate pesticides, which inhibit choline esterases, are detectable using a multi-enzyme inhibition assay (HPTLC-EI) based on rabbit liver esterase (RLE), Bacillus subtilis (BS2) esterase, or cutinase (from Fusarium solani pisi). Because choline esterase inhibition is more effective after conversion of thiophosphate thions into their corresponding oxons, a pre-oxidation step was added to the HPTLC-EI assay. Bromine vapour was found to be more effective than iodine or UV irradiation for oxidation. Following oxidation, the inhibitory strength of parathion, parathion-methyl, chlorpyrifos, chlorpyrifos-methyl, and malathion, expressed as HPTLC enzyme inhibition factors (f(i)), increased by approximately 2 orders of magnitude. In contrast, bromine oxidation of organophosphate and carbamate insecticides resulted in a slight reduction in their inhibition factors, due to partial bromination and degradation of the parent compounds, while bromine oxidation increased the inhibition factors for demeton-S-methyl and propoxur. Apple juice and water samples spiked with paraoxon (0.001 mg/L), parathion (0.05 mg/L), and chlorpyrifos (0.5 mg/L) were used to test the HPTLC-EI system, resulting in mean recoveries of 95-106% and 91-102% for RLE and cutinase, respectively.

Multi-enzyme inhibition assay for the detection of insecticidal organophosphates and carbamates by high-performance thin-layer chromatography applied to determine enzyme inhibition factors and residues in juice and water samples.

Esterase inhibition assays provide an effect-directed tool of rapid screening for inhibitors in environmental and food samples. According to a multi-enzyme microtiter-plate assay, rabbit liver esterase (RLE), Bacillus subtilis esterase (BS2), and cutinase from Fusarium solani pisi (CUT) were used for the detection of 21 organophosphorus and carbamate pesticides by high-performance thin-layer chromatography-enzyme inhibition assays (HPTLC-EI). Staining was performed with Fast Blue Salt B coupling to alpha-naphthol enzymatically released from the respective acetate used as substrate. Quantitative analysis was achieved by densitometric evaluation at 533 nm. Enzyme inhibition factors derived from HPTLC-EI were calculated from the slopes of the linear calibration curves, which allowed comparisons to published inhibition constants and well correlated to sensitivity parameters. Limits of detection ranged from a few pg/zone for organophosphates as strongest inhibitors to a few ng/zone for most carbamates, when RLE and BS2 were used. Without oxidation, chlorpyrifos and parathion were directly detectable at approximately 60 and 14 ng/zone, respectively. As the enzyme of lowest sensitivity, CUT was able to detect insecticides of high and low inhibitory power from the ng to microg range per zone. Due to high selectivity of enzyme inhibition, oxon impurities of thionophosphate standards were strongly detected, although only present in low traces. The exemplary application of HPTLC-EI (RLE) to apple juice and drinking water samples spiked with paraoxon (0.001 mg/L), parathion (0.05 mg/L) and chlorpyrifos (0.5mg/L) resulted in mean recoveries between 71 and 112% with standard deviations of 2.0-18.3%.