The recognition of high molecular weight melanoidins as the main components responsible for radical-scavenging capacity of unheated and heat-treated Canadian honeys

As the Maillard reaction is known to occur in heat-treated foods, unheated and heated honey samples were subjected to the activity-guided fractionation and size-exclusion chromatography to compare the degree of browning, radical scavenging activity (ORAC) and molecular size of the fractions obtained. Heat-treatment increased browning in the fractions of light- and medium-coloured honeys (p < 0.002), accelerated the formation of high molecular weight melanoidins (85–232 kDa) and significantly increased ORAC values (p < 0.0001). In contrast, melanoidin content and ORAC decreased in the fractions of heat-treated dark buckwheat honey (p < 0.001). Unheated dark honey contained a significantly higher amount of melanoidins than other honeys (p < 0.007). Together, results showed that at low initial concentration of melanoidins, heat-treatment accelerated formation of new melanoidins and increased ORAC, while at high concentration it caused decrease and a reduction of radical scavenging activity. This study emphasises the importance of non-enzymatic browning in the prediction of the antioxidant pool in thermally processed honeys.     

Influence of the action of exogenous enzymes on the polyphenolic composition of pea: Effect on the antioxidant activity

The polyphenolic composition of green pea (Pisum sativum, variety Esla) was determined by HPLC-DAD and HPLC-(ESI)MS procedures. The action of the enzymes tannase, α-galactosidase, phytase and viscozyme on pea flours, at a semi-pilot scale, was assayed for the polyphenolic composition. Different modifications in the initial concentration of polyphenolics occur depending on the enzyme, but a general decrease was observed in all of them. Free radical scavenging activity of the methanolic extracts, determined by the DPPH test, was also evaluated. The treatments with phytase, viscozyme, α-galactosidase or tannase produced a decrease in the antioxidant activity when compared to raw peas. The results of the analysis of principal components to examine the relationship between antioxidant activity (EC₅₀) and concentrations of polyphenolics in each of the pea samples showed that the hydroxycinnamic compounds are the most related to antioxidant activity.     

Valorisation of grape pomace by the extraction of phenolic antioxidants: Application of high voltage electrical discharges

The aim of this study was to optimise the electrically assisted extraction in order to obtain grape pomace extracts with high polyphenols content, which would be potentially interesting for applications as natural antioxidants. High voltage electrical discharges (HVED) were applied for intensification of the extraction. The effects of the energy input, the electrodes distance gap and the liquid-to-solid ratio were studied. Diffusion was then carried out in different mixtures of water and ethanol for one hour at 20, 30, 40 and 60 °C. The most efficient extraction was (i) an HVED pre-treatment at 80 kJ/kg with an electrodes distance of 5 mm in a liquid-to-solid ratio of 5 followed by (ii) a diffusion with 30% ethanol in water at 60 °C for 30 min. The highest total polyphenols content reached 2.8 ± 0.4 gGAE/100gDM with a corresponding antioxidant activity of 66.8 ± 3.1 gTEAC/kgDM. The polyphenols extraction rate increased with temperature in accordance to an Arrhenius type of relationship: activation energy of 21.5 ± 1.1 kJ/mol for HVED treated systems against 0.5 ± 0.2 kJ/mol for untreated ones.     

Antioxidant activity and chemical constituents of essential oil and extracts of Rhizoma Homalomenae

Antioxidant activity and composition of essential oil and extracts of Rhizoma Homalomenae were determined. The extracts, especially the ethyl acetate extract (QJ4 fraction) of the aqueous residue after oil distillation, had considerable antioxidant potency which was significantly associated with their total phenolic and flavonoid contents, but the essential oil showed only weak or moderate activity. GC–MS analysis of the essential oil (yield: 0.82%, v/w) resulted in the identification of 77 compounds, accounting for 96.5% of the content of the oil. The major components, epi-α-cadinol (14.8%), α-cadinol (14.8%), α-terpineol (13.8%), linalool (11.1%), terpinen-4-ol (4.92%), and δ-cadinene (4.91%) constituted 64.3% of it. LC–MS/MS and HPLC analyses showed seven phenolic compounds (protocatechuic acid, vanillic acid, syringic acid, caffeic acid, p-coumaric acid, ferulic acid and apigenin) with a great amount in the ethyl acetate extract (QJ4 fraction). The strong antioxidant properties of the plant extracts may be attributed to the presence of these phenolics.     

Analysis of flavonoids and phenolic acids in Greek aromatic plants: Investigation of their antioxidant capacity and antimicrobial activity

The phenolic fraction of plant extracts has been linked to their antioxidant capacity and antimicrobial activity. In the present paper, RP-HPLC with UV detection was employed to distinguish phenolic compounds from the other constituents of the plants examined. The phenolic substances were identified and quantified after comparison with reference standards. A GC–MS method is also presented for characterization of different phenolics as trimethylsilyl derivatives. The antioxidant capacity was determined, in dried plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically applying the Folin-Ciocalteu assay. It ranged from 2.9 to 28.2 mg gallic acid/100 g dry sample. Antimicrobial activity of the extracts against selected microorganisms was also investigated.     

Variation of glucosinolates in 62 varieties of Chinese cabbage (Brassica rapa L. ssp. pekinensis) and their antioxidant activity

Glucosinolate (GSL) and antioxidant activity in 62 varieties of Chinese cabbage (Brassica rapa L. ssp. pekinensis) were determined by HPLC and DPPH, HRSA, and FRAP assays. Five aliphatic GSLs: progoitrin, sinigrin, glucoalyssin, gluconapin, and glucobrassicanapin; four indolyl GSLs: 4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrassicin; one aromatic GSL: gluconasturtiin were identified. Glucobrassicanapin and gluconapin documented the most abundant (average 4.52 and 3.72 μmol/g DW, respectively). The contents of total GSLs varied extensively among 62 varieties (range from 2.83 to 48.53 μmol/g DW). Comprehensive differences in total and individual GSL contents have also been observed among different varieties. Indolyl and aromatic GSL together accounted 26% of the total GSLs; but there are few differences among varieties. FC7 and FI17 could be good candidates for future breeding programs since they had a high quantity of glucobrassicin (2.10 and 1.66 μmol/g DW, respectively). Most of the Chinese cabbage varieties showed significant antioxidant activities when compare with positive control. However, three antioxidant assays were not significantly correlated with total GSLs. The presence of significant quantities of glucobrassicin in some varieties should be studied more extensively, since GSL is the precursor of indole-3-carbinol, a potent anticancer isothiocyanate.     

Influence of conventional and organic agricultural practices on the phenolic content in eggplant pulp: Plant-to-plant variation

Consumer awareness, pesticide and fertiliser contaminations, and environmental concerns have resulted in significant demand for organically grown farm products. The present study evaluates the influence that organic and conventional farming practices exert on the total phenolic content in eggplant samples from two cultivars, Blackbell (American eggplant) and Millionaire (Japanese eggplant), grown by conventional and organic farming practices with similar climatic conditions. In addition, plant-to-plant variation in phenolic content was determined from eight eggplant samples collected from different plants grown at various field positions. Samples were separately assayed for phenolic content by the two independent procedures, HPLC/LC–MS and Folin–Ciocalteu assay. The results of the HPLC analysis showed significant plant-to-plant variation (% RSD ranged from 22% to 48%) in 5-caffeoylqunic acid content (the most abundant phenolic acid) in eight eggplant samples collected from eight different plants belonging to the same cultivar and grown under similar conditions. The cv. Millionaire showed a higher total phenolic content compared to the Blackbell cultivar. We did not observe significant consistent trend in the phenolic content of eggplant samples grown with organic and conventional farming practices with both eggplant cultivars. We, however, observed a strong correlation (R² = 0.87) between total phenolic content as assayed by Folin–Ciocalteu assay and 5-caffeoylqunic acid, a major phenolic acid in eggplant extract. This study concludes that multiple repetitive analyses of plant products collected from different plants grown over various time periods (seasons) at different locations should always be carried to unambiguously prove the impact of growing conditions on phenolic content or antioxidant activity.     

Low molecular weight chitosan prepared with the aid of cellulase,lysozyme and chitinase: Characterisation and antibacterial activity

A continuous set of low molecular weight chitosan (LMWC) products was successfully made for this study by coordinating three enzymes (chitinase, lysozyme and cellulase) and two different deacetylated chitosan substrates (80% and 92%). It was observed that the intrinsic viscosity molecular weight (M_V) and SEC-HPLC-determined M_W distribution of LMWC were directed by both the used enzyme and the degree of chitosan substrate acetylation. LMWC prepared using lysozyme and 92%-deacetylated chitosan had larger M_W and, therefore, possessed higher antibacterial activity, as compared to other combinations. LMWC enzyme-directed properties suggest chitinase is more predictable and flexible to produce the specified M_V of LMWC. LMWC’s solubility and antibacterial activity, determined as minimum inhibitory concentration (MIC), against Escherichia coli exhibited a negative linear relationship with log M_V. E. coli strains showed much higher susceptibility to LMWCs than Staphylococcus aureus strains did. Both species also showed intra-species sensitivity diversity toward LMWC.     

Low molecular weight serine protease from the viscera of sardinelle (Sardinella aurita) with collagenolytic activity: Purification and characterisation

A new low molecular weight (LMW) serine-protease from sardinelle (Sardinella aurita) viscera was purified using ammonium sulphate precipitation and Sephadex G-100 gel filtration, with a 3.82-fold increase in specific activity. The molecular weight of the enzyme was estimated to be 14.2 kDa by SDS-PAGE. The optimum pH and temperature for the enzyme activity were around pH 8.0 and 60 °C, respectively. The purified protease was strongly inhibited by phenylmethylsulphonyl fluoride, a serine-protease inhibitor, and soybean trypsin inhibitor. The N-terminal amino acid sequence of the first 10 amino acids of the purified protease was APVQPCVVVI. This sequence showed low homology with several peptidases, suggesting that the enzyme is a new protease. Interestingly, the protease was found to cleave collagen type I and hydrolyze succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide (sAAPFpna), an amide substrate of chymotrypsin. Our findings indicate that the S. aurita protease is a new LMW enzyme with collagenolytic activity.