Black bean (Phaseolus vulgaris L. cv. "Tolosa") polyphenolic composition through cooking and in vitro digestion.

In this study, five different black bean (Phaseolus vulgaris L. cv. Tolosa) populations cultivated in different geographical areas including Oiartzun, Andoain, Azkoitia, San Esteban and Amasa Villabona, were studied and their polyphenolic content was determined. Two food products were prepared from the five different bean populations, cooked "Tolosa" beans and a hummus made with "Tolosa" cooked beans. Moreover, the variations of total polyphenol content (TPC), total anthocyanin content (TAC) and free radical scavenging activity by the 1,1-diphenyl-1-picrylhydrazyl (DPPH) method were analyzed for raw beans, cooked beans, and "Tolosa" beans hummus. Polyphenolic detailed composition was determined by means of HPLC-MS-QTOF analysis. The "Tolosa" bean population richest in polyphenols was selected in order to study the effect of in vitro digestion on the polyphenolic content and antioxidant effect and the degradation of the main anthocyanins was followed during the in vitro digestion. Finally, the effect of the different phases of digestion on the cytotoxicity in Caco-2 cells was determined. The results suggest that cooking "Tolosa" black beans results in an increase in the bioaccessibility of polyphenols and their antioxidant activity, which, additionally has a positive effect on Caco-2 intestinal cell viability.

Study on the phenolic composition and antioxidant properties of white-, yellow-, and black-corn (Zea mays L.) foodstuffs.

BACKGROUND: Black corn possesses potent antioxidants, but these are mostly lost during processing. In this study we evaluated the antioxidant content of two different black ('Millo Corvo') corn-based products (i.e. tortillas and cookies) subject to moderate processing. A parallel study on white and yellow corns was carried out for comparison. RESULTS: Raw 'Millo Corvo' flour exhibited higher contents of phenolic acids, flavonoids, and particularly anthocyanins than white and yellow flours did. Phenolic acids decreased in cookies but did not in tortillas; flavonoids did not exhibit a clear tendency, and anthocyanins were always preserved. Antioxidant activity (AA) obtained for 'Millo Corvo' samples was twice as high as the value measured in white and yellow corns in terms of 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. The difference in AA was even more remarkable in terms of photochemiluminiscence antiradical activity. The conditions used during the cookie-making process enabled the natural antioxidant characteristics of 'Millo Corvo' to be preserved. However, the conditions applied to prepare tortillas resulted in major losses. CONCLUSIONS: The initial phenolics, in particular anthocyanins, and AA of 'Millo Corvo' flour can be maintained during processing as long as the conditions applied are 183 °C for 20 min in the absence of a high water content. Millo corvo products possess improved antioxidant characteristics compared with those from white and yellow corns. Millo corvo is a promising black corn type to prepare health-promoting corn-based foodstuffs. © 2023 Society of Chemical Industry.

Varietal Effect on Composition and Digestibility of Seedless Table Grapes (Vitis vinifera L.) under In Vitro Conditions.

Grapes are one of the richest sources of polyphenols in the Mediterranean diet and, therefore, a very good source of antioxidants in the human diet. For practical reasons, in recent years the market for seedless grape varieties has grown exponentially. These varieties are not well characterized yet, and therefore it is necessary to study the changes in composition that these new varieties bring in. Likewise, the effect of digestion on the bioavailability of polyphenols in foods of plant origin is well known, which, consequently, will also affect antioxidant activity and, in general, bioactivity. In this work, we studied the effect of the grape variety on the initial grape composition and on the absorbable fraction, as it would reach the intestine after in vitro digestion. Our results showed that black and red varieties have great potential for increasing the antioxidant content of the diet. Additionally, we have concluded that all polyphenols, with the exception of flavanols, see their bioavailable fraction diminished under in vitro conditions.

Effect of Baking Temperature on the Phenolic Content and Antioxidant Activity of Black Corn (Zea mays L.) Bread.

Black corn is known for its health-promoting properties, which are due to its high content of bioactive phytonutrients. However, the high temperatures required during the processing of bakery products usually trigger thermal degradation, and therefore, the loss of all labile bioactive compounds. In the present study, we evaluated the effect of baking temperature on the phenolic content (i.e., TPC, TAC and individual phenolics) and antioxidant activity in black corn (Millo corvo variety) bread. As a result, baking always resulted in a general decrease in TPC, even at 150 °C. In contrast, TAC only decreased when temperatures as high as 180 °C were applied. Some relevant individual phenolics were preserved during the whole process as long as 150 °C was used. In particular, the content of the major anthocyanin, namely, cyanidin-3-O-glucoside, hardly decreased from the raw flour to the final bread. The loss of antioxidant activity of Millo corvo raw flour during bread baking was avoided by heating at 150 °C. These results demonstrate the appropriate temperature to bake Millo corvo corn bread without losing the antioxidant characteristics and health-promoting properties of the starting black corn.

Enrichment of Olive Fruits in Antioxidant Content by Pre-Harvest Salicylic Acid Treatment.

We here study the effect of the pre-harvest application of salicylic acid at two different concentrations on the olive phenolic composition. Influence of the cultivar and harvesting day were considered. As a result, the total phenol content increased significantly, particularly when using 200 mg mL-1 of salicylic acid. However, the free radical scavenging activity was cultivar dependent. For instance, when the olives were harvested on day 3 and treated with 200 mg mL-1 of salicylic acid, the antioxidant activity decreased from 161 to 278 µg mL-1 for Arbequina, whereas it increased from 397 to 258 µg mL-1 for Picual. Generally speaking, oleuropein and hydroxytyrosol contents enhanced with the application of 200 mg mL-1 of salicylic acid. The results found suggest that exogenous salicylic acid is an interesting agronomic practice to enrich olive fruits in antioxidants.

Exogenous Salicylic Acid Improves Phenolic Content and Antioxidant Activity in Table Grapes.

Grapes contain high contents of phenolics, which are known to possess health promoting properties. Exogenous application of phytoregulators, mainly methyl jasmonate and abscisic acid, to grapevines to enhance phenolic content has been reported (Portu et al. Sci Hortic 240: 378-386, 2018; Ranjbaran et al. J Faculty Agric Kyushu Univ 56: 263-267, 2011). However, these phytohormones possess some drawbacks that can be overcome by using other phytoregulators as an alternative. In this work the effect of an additional phytohormone, salicylic acid, to grapevines on the phenolics and antioxidant activity of grapes was investigated. To our knowledge, salicylic acid has been earlier applied to grapevines to affect grape ripening and quality (Lóay. Egyptian J Basic Applied Sci 4: 227-230, 2017). However, this is the first time it is applied to increase the total phenolic content. As a result of our study, total phenol content and the free radical scavenging activity increased with 100 mg l-1 of salicylic acid. In particular, the total phenol content increased from 768.3 to 1843.5 mg 100 g-1 and the IC50 values decreased from 45.2 to 13.2 mg ml-1. Also the contents of individual phenolics mostly increased significantly with 100 mg l-1 of salicylic acid, except anthocyanins. Higher concentrations of salicylic acid (ie, 500 mg l-1vs 100 mg l-1) did not result in higher contents of phenolics. Therefore, 100 mg l-1 was selected as the best salicylic acid concentration to be used in the treatment. The application of exogenous salicylic acid to grapevines is an interesting agronomic practice to obtain table grapes with improved health-promoting properties.

Effect of postharvest methyl jasmonate treatment on fatty acid composition and phenolic acid content in olive fruits during storage.

BACKGROUND: The nutritional effects of both table olives and olive oil are attributed not only to their fatty acids but also to antioxidant phenolics such as phenolic acids. Delays in oil processing usually result in undesirable oxidation and hydrolysis processes leading to formation of free fatty acids. These alterations create the need to process oil immediately after olive harvest. However, phenolic content decreases drastically during olive storage resulting in lower quality oil. In the present study we propose postharvest methyl jasmonate treatment as a mean to avoid changes in fatty acid composition and losses of phenolic acids during olive storage. RESULTS: Contents of fatty acids and phenolic acids were estimated in methyl jasmonate treated olives throughout 30-day storage, as compared with those of untreated olives. Significant decreases of saturated fatty acids were observed in treated samples whereas increases of oleic, linoleic and linolenic acids were respectively measured (i.e. from 50.8% to 64.5%, from 7.2% to 9.1% and from 1.5% to 9.3%). Also, phenolic acid contents increased significantly in treated olives. Particularly, increases of gallic acid from 1.35 to 6.29 mg kg-1 , chlorogenic acid from 9.18 to 16.21 mg kg-1 , vanillic acid from 9.61 to 16.99 mg kg-1 , caffeic acid from 5.12 to 12.55 mg kg-1 , p-coumaric acid from 0.96 to 5.31 mg kg-1 and ferulic acid from 4.05 to 10.43 mg kg-1 were obtained. CONCLUSION: Methyl jasmonate treatment is proposed as an alternative postharvest technique to traditional methods to guarantee olive oil quality when oil processing is delayed and olive fruits have to necessarily to be stored. © 2016 Society of Chemical Industry.

Supercritical fluid extraction of grape seeds: extract chemical composition, antioxidant activity and inhibition of nitrite production in LPS-stimulated Raw 264.7 cells.

Grape by-products are a rich source of bioactive compounds having broad medicinal properties, but are usually wasted from juice/wine processing industries. The present study investigates the use of supercritical fluid extraction (SFE) for obtaining an extract rich in bioactive compounds. First, some variables involved in the extraction were applied. SFE conditions were selected based on the oil mass yield, fatty acid profile and total phenolic composition. As a result, 40 °C and 300 bar were selected as operational conditions. The phenolic composition of the grape seed oil was determined using LC-DAD. The antioxidant activity was determined by ABTS and DPPH assays. For the anti-inflammatory activity the inhibition of nitrite production was assessed. The grape seed oil extracted was rich in phenolic compounds and fatty acids with significant antioxidant and anti-inflammatory activities. From these results, added economic value to this agroindustrial residue is proposed using environmentally friendly techniques.

Phenylalanine ammonia-lyase, flavanone 3ß-hydroxylase and flavonol synthase enzyme activity by a new in vitro assay method in berry fruits.

An HPLC method for the determination of phenylalanine ammonia-lyase, flavanone 3ß-hydroxylase and flavonol synthase enzyme activity is proposed. This method is based on the determination of the compounds produced and consumed on the enzymatic reaction in just one chromatographic analysis. Optimisation of the method considered kinetic studies to establish the incubation time to perform the assay. The method here described proved to be an interesting approach to measure the activities of the three enzymes simultaneously increasing the rapidity, selectivity and sensitivity over other exiting methods. The enzyme activity method developed was applied to strawberry, raspberry, blackberry, redcurrant and blackcurrant fruits.

Isolation of the four methyl jasmonate stereoisomers and their effects on selected chiral volatile compounds in red raspberries.

The four stereoisomers present in a commercial sample of methyl jasmonate (MJ) were isolated at semi-preparative scale by HPLC, using a permethylated ß-cyclodextrin column. This allowed the baseline resolution and collection of both major (methyl jasmonates) and minor (epi-methyl jasmonates) stereoisomers. When 1.5 mL of a 5mg per mL MJ solution were injected, isolated amounts were 3.56 mg for (-) and (+)-methyl jasmonates, with respective purities of 96.1% and 99.9%, and 0.18 mg for (-)- and (+)-epi-methyl jasmonates, with 98.6% and 91.6% respective purities. The post-harvest treatment of red raspberry fruits with the pure methyl jasmonate stereoisomers isolated proved that (-)-epi-MJ either promotes the bioformation of (+)-limonene or inhibits that of (-)-limonene to a greater extent than the other three MJ stereoisomers, while the biosynthesis of the (+)-enantiomer of α-ionone is favoured equally, whichever MJ stereoisomer used. The results obtained in the present study might be used to obtain food products with improved sensory characteristics.

A new approach based on off-line coupling of high-performance liquid chromatography with gas chromatography-mass spectrometry to determine acrylamide in coffee brew.

A new method based on off-line coupling of LC with GC in replacement of conventional sample preparation techniques is proposed to analyze acrylamide in coffee brews. The method involves the preseparation of the sample by LC, the collection of the selected fraction, its concentration under nitrogen, and subsequent analysis by GC coupled with MS. The composition of the LC mobile phase and the flow rate were studied to select those conditions that allowed separation of acrylamide without coeluting compounds. Under the conditions selected recoveries close to 100% were achieved while LODs and LOQs equal to 5 and 10 µg/L for acrylamide in brewed coffee were obtained. The method developed enabled the reliable detection of acrylamide in spiked coffee beverage samples without further clean-up steps or sample manipulation.

Evaluation of pesticide residue contents in fruit juice by solid-phase microextraction and multidimensional gas chromatography coupled with mass spectrometry.

A new method based on the use of SPME followed by the MDGC-MS analysis was developed to determine pesticides in fruit juice. Different pesticide mixture standards (i.e. Mix 101, 13 and 164) were initially analyzed to optimize the separation conditions. To evaluate the advantages of the two-dimensional system over monodimensional GC, a comparative study on relative standard deviations, detection limits and correlation coefficients was carried out. As a result, selective transfers of some pesticides from the first to the second dimension were at times essential to avoid overlapping. The selected separation conditions from the study with standards were applied to fruit juices spiked with some pesticide standards. The results found in this work prove that the employment of a multidimensional analysis technique permits to avoid false positives obtained frequently on monodimensional techniques as a consequence of interferences of the analytes with matrix components.

(+)-methyl jasmonate-induced bioformation of myricetin, quercetin and kaempferol in red raspberries.

The effect of postharvest treatment with enantiomers of methyl jasmonate (MJ) in conjunction with ethanol on bioformation of myricetin, quercetin and kaempferol in red raspberry was studied. For comparison, postharvest treatment with the commercial stereoisomeric mixture of MJ in conjunction with ethanol was simultaneously accomplished. The levels obtained were contrasted with those determined in untreated (control) samples. Exogenous (+)-MJ induced an enhancement in the levels of myricetin, quercetin and, particularly, kaempferol whereas the exposition to (-)-MJ exhibited the opposite effect. Enzymatic assays were carried out in presence and absence of (-)-MJ and (+)-MJ to evaluate possible changes in the activity of the enzymes regulating the bioformation of flavonols in red raspberries as a consequence of the treatments. From the results of the assays both (-)-MJ and (+)-MJ inhibited the activity of flavanone 3ß-hydroxylase (FHT) and flavonol synthase (FLS), which are directly involved in the formation of flavonols from (-/+)-naringenin. From these results, it is speculated that the activity of phenylalanine ammonia lyase (PAL) regulating the formation of (-/+)-naringenin from l-phenylalanine by (+)-MJ in conjunction with ethanol is promoted. Postharvest treatment of red raspberry with (+)-MJ in ethanol is proposed as a mean to increase flavonol content in red raspberries.

Exogenous methyl jasmonate diminishes the formation of lipid-derived compounds in boiled potato (Solanum tuberosum L.).

BACKGROUND: During potato storage the tubers tend to develop off-flavours, mainly due to lipid-derived aldehydes, whose formation is increased after boiling or processing. This may become a problem when boiled or precooked potatoes are used. Methyl jasmonate (MJ) is a phytohormone capable of modifying plant and food composition. The aim of this work was to control the formation of off-flavours in different cultivars of raw potato during storage by MJ treatment. RESULTS: The use of Carbowax 20M as support in MJ emulsion resulted in significantly (P < 0.05) lower levels of lipid-derived compounds than the use of lanolin. Levels of the lipid-derived aldehydes propanal, hep-2-enal, nonanal, oct-2-enal and hexanal, although dependent on variety, generally decreased significantly (P < 0.05) after treatment with MJ/Carbowax 20M emulsion. Sugar-derived compounds such as pyridine and furfural, however, were not affected by this treatment. CONCLUSION: Treatment with MJ/Carbowax 20M can be used to diminish the formation of off-flavours in boiled potato. However, this treatment is not useful to avoid the sweetening caused by sugar-derived products. Additional advantages of exogenous MJ are a delay in the appearance of sprouting and aging and hence prolongation of the storage life of potato tubers.

Impact of postharvest methyl jasmonate treatment on the volatile composition and flavonol content of strawberries.

BACKGROUND: Although strawberry aroma is very complex, certain compounds have been described as main contributors, i.e. furanones, aldehydes, alcohols, sulfur compounds and particularly methyl and ethyl esters. In addition, strawberries possess potent antioxidant activity because of their high content of phenolic compounds. Among them, flavonols are highlighted as important antioxidant compounds in strawberry. The objective of this study was to assess the effect of methyl jasmonate (MJ) on the composition of the major contributors to aroma and on the content of certain flavonols in strawberry fruits. RESULTS: The levels of all studied volatile compounds were significantly affected by MJ treatment, though the individual effect differed according to the specific compound considered. Most of them increased significantly (P < 0.05), except methyl butanoate, which always showed higher levels in untreated strawberries. In contrast to aroma compounds, the change in the concentration of flavonols (i.e. myricetin, quercetin and kaempferol) was not significant in MJ-treated strawberries. Considering the health-promoting activity of these compounds, further investigations on the experimental conditions related to the treatment are required to control flavonol bioformation by means of MJ. CONCLUSION: The exogenous application of MJ vapour to strawberry enhances, in general, the production of the most relevant aroma-active compounds. On the contrary, MJ treatment does not appear to influence the levels of myricetin, quercetin and kaempferol. Thus postharvest MJ treatment is proposed as an approach to obtain improved strawberry fruits in terms of sensory quality and health-promoting properties.

Development of a method based on on-line reversed phase liquid chromatography and gas chromatography coupled by means of an adsorption-desorption interface for the analysis of selected chiral volatile compounds in methyl jasmonate treated strawberries.

A method based on the use of the through oven transfer adsorption-desorption (TOTAD) interface in on-line coupling between reversed phase liquid chromatography and gas chromatography (RPLC-GC) for the determination of chiral volatile compounds was developed. In particular, the method was applied to the study of the influence of methyl jasmonate (MJ) treatment on the production and enantiomeric composition of selected aroma compounds in strawberry. The compounds studied were ethyl 2-methylbutanoate, linalool and 4-hydroxy-2,5-dimethyl-3(2H)-furanone (i.e. furaneol), which were examined on days 3, 6 and 9 after treatment. The method developed resulted in relative standard deviations (RSDs) of 21.6%, 8.1% and 9.8% and limits of detection (LD) of 0.04, 0.07 and 0.02mg/l for ethyl 2-methylbutanoate, linalool and furaneol, respectively. The application of the RPLC-TOTAD-GC method allowed higher levels of ethyl 2-methylbutanoate, linalool and furaneol to be detected, particularly after 9 days of treatment. Besides, MJ demonstrated to affect the enantiomeric distribution of ethyl 2-methylbutanoate. On the contrary, the enantiomeric composition of linalool and furaneol kept constant in both control and MJ-treated strawberries throughout the study. These results are discussed.

Jasminum polyanthum Franch. as a natural source of (-)-methyl jasmonate: an alternative to the use of the synthetic standard.

INTRODUCTION: Methyl jasmonate (MJ) contains two chiral centres at C-3 and C-7 in its chemical structure, which implies that it can exist in four possible stereoisomeric forms, namely (+)-MJ, (-)-MJ, (+)-epiMJ and (-)-epiMJ. The absolute configuration of the two side chains of MJ affects the biological activity associated with this compound. OBJECTIVE: To isolate pure (-)-MJ from a natural source, Jasminum polyanthum Franch., with the intention of increasing the knowledge about its biological properties, including its effect on the biosynthesis of plant metabolites. METHODOLOGY: The method used was based on steam distillation extraction (SDE) as an extraction technique followed by high-performance liquid chromatography (HPLC) as a purification procedure. The HPLC flow-rate as well as the number of fractions accumulated were optimised to achieve the concentration and purity required. RESULTS: The employment of 0.3 mL/min as HPLC flow-rate and the accumulation of three HPLC fractions allowed the required enantiomeric purity (95%) and concentration (0.36 mg/L in each HPLC fraction) to efficiently obtain (-)-methyl jasmonate from Jasminum polyanthum Franch. to be achieved. CONCLUSION: The approach proposed may enable the properties and effect of pure (-)-MJ on plant responses to be studied. The use of a natural source to obtain (-)-MJ is presented as an alternative to the enantioselective synthesis and enantiomeric resolution from the standard racaemic mixture.

Enantioselective isolation of methyl jasmonate using permethyl-beta-cyclodextrin HPLC.

A method based on the use of HPLC for the enantioselective resolution of the four stereoisomers of methyl jasmonate (MJ) with no need for the previous formation of the diastereoisomers is developed. To that end, a Nucleodex-beta-PM column as well as an optimization process considering different flow rates and mobile phase compositions were required. As a result, 0.8 mL/min and 55:45 methanol/water composition were the conditions selected to carry out the separation of the stereoisomers. Isolation of pure (-)- and (+)-MJ was accomplished by collecting the HPLC fractions corresponding to their elution time. SPE was subsequently used to concentrate and change the solvent of the HPLC fractions collected. Chiral GC and polarimetry were additionally employed to evaluate the purity and optical rotation, respectively, of the enantiomers separated. The results found in this study are particularly relevant considering that MJ stereoisomers are not commercially available.

Online RPLC-GC via TOTAD method to isolate (+)-methyl epijasmonate from lemon (Citrus limon Burm.).

Pure (+)-methyl epijasmonate was isolated from lemon ( Citrus limon Burm.) for the first time. To that aim, two commercial essential oils and one homemade extract were included in the present paper. First, a study on the appropriate chromatographic conditions to avoid the epimerization from methyl epijasmonate to the more stable methyl jasmonate was accomplished. The results obtained are discussed. The presence of (+)-methyl epijasmonate in the three samples studied was initially established through the direct injection into GC-MS. However, the overlapping of (+)-methyl epijasmonate with other matrix components made it necessary to employ a multidimensional technique. RPLC-GC analysis via through-oven transfer adsorption-desorption (TOTAD) provided the selectivity and sensitivity required, reflecting that the homemade lemon extract was an adequate natural source to obtain pure (+)-methyl epijasmonate by means of the collection of the corresponding RPLC fraction.

Through oven transfer adsorption-desorption interface for the analysis of methyl jasmonate in aromatic samples by on-line RPLC-GC.

A fully automated method for the determination of medium volatility compounds in aromatic samples was developed. Specifically, the determination of methyl jasmonate in jasmine fragrances was performed by using the through oven transfer adsorption-desorption (TOTAD) interface for the on-line coupling between RPLC-GC. A study of the most relevant variables involved in the performance of the TOTAD interface for medium volatility compounds was carried out by testing different values of helium flow (100, 300, 400, and 500 mL/min), transfer speed (0.1, 0.3, 0.5, and 2.0 mL/min), and methanol/water percentages (86:14, 85:15, 83:17, 80:20, and 70:30). The method developed provided satisfactory repeatability (RSD for retention times of 0.15% and for peak areas of 9.4%) and recovery (71%) as well as excellent LOD (0.01 mg/L) for methyl jasmonate in commercial jasmine essence under the experimental conditions selected as optimum. Additional advantages of the automated RPLC-TOTAD-GC method proposed in the present work are its rapidness, reliability, and the possibility of directly introducing the sample with no further pretreatment.