Pressurized acidified water extraction of black carrot [<Emphasis Type="Italic">Daucus carota</Emphasis> ssp. <Emphasis Type="Italic">sativus</Emphasis> var. <Emphasis Type="Italic">atrorubens</Emphasis> Alef.] anthocyanins

The anthocyanins present in black carrot were extracted with pressurized water acidified with sulfuric, citric and lactic acids. Anthocyanin degradation became significant above 100 °C and there was no improvement when extraction pressure was increased to 100 bar. Therefore, the extraction from black carrot was carried out at temperatures 50, 75 and 100 °C under 50 bar pressure. The extraction efficiencies in terms of acylated and non-acylated anthocyanins were comparable for all three acids used to acidify water at 50 °C, while similar results were observed at 75 °C for both citric and lactic acids. Water acidified with lactic acid showed significantly higher extraction efficiency at 100 °C compared to water acidified with sulfuric and citric acids. Highest degree of polymerization together with increasing degree of browning was observed within extracts when sulfuric acid was used. On the other hand, when organic acids were used to acidify water, a higher extraction efficiency of anthocyanins, accompanied with a relatively low polymerization and browning was observed, with lactic acid giving the best results.     

Quantification of anthocyanins in black carrot extracts (<Emphasis Type="Italic">Daucus carota</Emphasis> ssp.<Emphasis Type="Italic"> sativus</Emphasis> var.<Emphasis Type="Italic"> atrorubens</Emphasis> Alef.) and evaluation of their color properties

Pigment composition of 15 black carrot cultivars (Daucus carota ssp. sativus var. atrorubens Alef.) was screened by HPLC-MS. Up to seven cyanidin glycosides, five of which were acylated with hydroxycinnamic and hydroxybenzoic acids, were identified and quantified in the roots by HPLC-DAD. Contents of individual compounds indicated great differences in the potential of anthocyanin accumulation both between different cultivars and carrots of the same cultivar. Total anthocyanin amounts ranged from 45.4 mg/kg dry matter to 17.4 g/kg dry matter. To the best of our knowledge, this is the first report on the quantification of individual anthocyanins in roots of different black carrot cultivars. The determination of color properties in the extracts under various pH conditions proved black carrot anthocyanins to be applicable as natural food colorants also for low-acid food commodities, whereas a considerable loss of color was noted under nearly neutral conditions. Additionally, relatively high saccharide contents were found in almost all cultivars which may be disadvantageous when coloring concentrates are produced from carrot roots.     

Optimization extraction of anthocyanins from purple corn (<Emphasis Type="Italic">Zea mays</Emphasis> L.) cob using tristimulus colorimetry

Optimization of the conditions for extracting anthocyanins from the cob, a byproduct of purple corn (Zea mays L.), was investigated in this paper. A full factorial design (2² × 3²) was used. The factors studied were type of solvent: ethanol or methanol; solvent/water mixture: 100, 90, 80% v/v; type of acid: citric acid or acetic acid; and concentrations of acid: 0.25, 0.5, 1% v/v. Tristimulus colorimetry was employed to evaluate the yield and quality of anthocyanins extracts. The results showed that the maximum yield (5.90 mg/g dw) was obtained with the combination of 80% (v/v) methanol and 1% (v/v) citric acid, and that the relatively high chroma (C* = 23.60) and hue angle (h = 16.63) values of the anthocyanins extract were observed under the same conditions. Eight kinds of anthocyanins in the cob were detected by HPLC–DAD, most the anthocyanins were non-acylated.     

Factorial design optimisation of grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis>) seed polyphenol extraction

A 2³-full factorial design and response surface methodology were deployed to assess some basic factors (time, % ethanol and pH) affecting profoundly the extractability of polyphenolic phytochemicals from grape (V. vinifera) seeds. In an effort to obtain a thorough insight into the applicability of the models established, seed extracts from three different varieties were tested, by determining several indices of the polyphenolic composition, such as total polyphenol (TP), total flavanol (TFl) and proanthocyanidin (PC) concentrations. It has been shown that the models generated can adequately predict the recovery levels for each polyphenol group, but the optimal conditions predicted for TP, TFl and PC recovery varied significantly. Notable differences were also seen among the different varieties. Correlation of the polyphenol indices with the antiradical activity and reducing power of the extracts indicated that there is no consistent pattern to associate specific polyphenol classes with the antioxidant potential, which might be an evidence of antagonism. Examination of the optimally obtained extracts using liquid chromatography-mass spectrometry revealed that the most prominent compounds were catechin, some flavanol dimers and galloylated derivatives thereof.     

Phenolic compounds content and antioxidant activity in pomace from selected red grapes (Vitis vinifera L. and Vitis labrusca L.) widely produced in Brazil

The phenolic compounds content and antioxidant activity of pomace from the vinification of grape varieties widely produced in Brazil (Cabernet Sauvignon, Merlot, Bordeaux and Isabel) were investigated with a view to their exploitation as a potential source of natural antioxidants. Cabernet Sauvignon grape pomace was found to have the highest content of total phenolic compounds (74.75 mg gallic acid equivalent (GAE)/g), the highest antioxidant activity (determined using the 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging methods; 485.42 and 505.52 μMol Trolox equivalent antioxidant capacity (TEAC)/g, respectively), and the highest reducing power (determined using the FRAP method; 249.46 μMol TEAC/g). The Bordeaux variety showed the highest oxidation inhibition power (41.13%), determined using the β-carotene/linoleic acid method and the highest content of total anthocyanins (HPLC; 29.17 mg/g). Catechin was the most abundant non-anthocyanic compound identified in the grape pomace (150.16 mg/100 g) for all varieties. In this study, pomaces of the red wine vinification of Cabernet Sauvignon and Bordeaux varieties showed the highest potential as a source of antioxidant compounds and natural colourants, respectively.     

Phenolic compounds in Imburana (<Emphasis Type="Italic">Amburana cearensis)</Emphasis> powder extracts

In response to the increasing demand for aged distilled spirits, wood extracts have been investigated as accelerating agents in distilled spirits. The aim of this work was to obtain and to evaluate the potential use of Amburana cearensis powder extracts as an accelerating aging agent in sugar cane distilled spirits. Wood powder was submitted to different toasting treatments using conventional drying oven and microwave oven. Instead of using the traditional reflux extraction method, the phenolic compounds were extracted by means of ultrasonic treatment using a 50% ethanolic solution with pH adjusted to 4.5. The concentration of gallic acid, 5-hydroxymethylfurfural, furfural, vanillic acid, syringic acid, vanillin, coniferaldehyde, sinapaldehyde and coumarin, determined by high performance liquid chromatography (HPLC) were used as quality parameters of powder toasting treatment and ethanolic extraction. The use of toasted wood powder combined with ultrasonic extraction showed higher yields when compared to the traditional solvent extraction method when obtaining wood extracts. The extraction method presented in this work is also less time consuming.     

Evaluation of pathogenesis-related protein content and protein instability of seven white grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) clones from Casablanca Valley,Chile

Thaumatin-like proteins and chitinases are the main pathogenesis-related (PR) proteins found in grapes, grape juice and wine and are responsible for protein haze formation in bottled white wine during storage and transport. We have studied the effect of the content of both thaumatin-like proteins and chitinases on protein instability of Sauvignon blanc (clones 1, 107 and 242) and Chardonnay (clones 4, 5, 75 and Mendoza) grape juices from both a warm and a cold production zone in the Casablanca Valley, Chile. The PR proteins were identified and quantified by reversed-phase liquid chromatography (RP-HPLC). Protein instability was determined using a heat test and was expressed in nephelometric turbidity units (NTUs). Thaumatin-like (TL) proteins were identified as the major PR proteins present in all grape juices studied. Three TL proteins were identified and named Vitis vinifera thaumatin-like proteins 1, 2 and 3 (VVTL1, VVTL2 and VVTL3). Chitinase A (ChitA) was identified in the Sauvignon blanc and Chardonnay grape juices, and chitinase B (ChitB) was found only in Chardonnay grape juices. Significant differences in the protein content and in the type of protein were observed between grapes from different production zones and between grapes of different varieties, respectively.     

Isolation and structure elucidation of phenolic compounds in Chinese olive (<Emphasis Type="Italic">Canarium album</Emphasis> L.) fruit

Chinese olive (Canarium album L.), one native and well-known tropical fruit tree in the southeast of China, contain a large amount of phenolics and possess great pharmacological activities. In this study, phenolics were extracted from Chinese olive fruit using 80% (v/v) aqueous acetone, and seven phenolic compounds were isolated and purified by Polyamide column and Toyopearl HW-40 column chromatography from crude extracts. Their structures were elucidated by high performance liquid chromatography coupled to diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS), and where possible by ¹H NMR and ¹³C NMR spectrometry. Except gallic acid and hyperin, five phenolic compounds including methyl gallate, ethyl gallate, corilagin, kaempferol-3-glucoside and amentoflavone were first identified in Chinese olive.     

Optimization for encapsulation of green tea (<Emphasis Type="Italic">Camellia</Emphasis><Emphasis Type="Italic">sinensis</Emphasis> L.) extract by spray drying technology

Green tea has been credited with providing a wide variety of health benefits like higher total antioxidant, cancer prevention, and anti-irritant etc. Microencapsulation technology is used to prevent antioxidant loss during processing which involves entrapment of active material into carrier material. The experiments were designed using response surface methodology based on three level two factor (green tea extract, 10–25 % and drying inlet air temperature, 120, 130 and 140°C) central composite design. The feed emulsion was prepared with green tea extract as core material and 40 % maltodextrin concentration as wall material with the ratio of core to wall (1:2) and it was fed into the spray dryer at varied drying inlet air temperature to get encapsulated green tea extract powder. The quality characteristics were analysed such as moisture content, total antioxidant activity and total phenolic content and the process conditions were optimized using desirability function methodology. At optimum spray drying condition moisture content, total antioxidant activity and total phenolic content were found to be 3.1 % w.b., 72.91 % free radical scavenging activity by DPPH method, 57.81 mg/g of dry matter, expressed as gallic acid equivalent respectively with desirability value of 0.92.     

Effect of glasswort (<Emphasis Type="Italic">Salicornia herbacea</Emphasis> L.) and rice (<Emphasis Type="Italic">oryza sativa</Emphasis> L.) on quality of <Emphasis Type="Italic">Ganjang</Emphasis> (Korean Soy Sauce)

Four types of meju were made from 100%(w/w) defatted soybean (DFS), a mixture of 80%(w/w) defatted soybean, and 20%(w/w) glasswort (DFS-G), a mixture of 80%(w/w) defatted soybean and 20%(w/w) rice (DFS-R), and a mixture of 60%(w/w) defatted soybean, 20%(w/w) glasswort, and 20%(w/w) rice (DFS-GR). Four types of Korean traditional soy sauce were prepared from the 4 types of meju. Mineral and antioxidant contents in the soy sauce made of DFS-G and DFS-GR were significantly higher than others. Citric, malic, succinic, lactic, and pyroglutamic acid contents in soy sauce made of DFS-R and DFS-GR were 1.3–1.5 times higher than others. Total nitrogen and free amino acid contents in soy sauce were correlated with DFS concentration in the meju. The bacterial community in the non-fermented meju-making ingredients was replaced largely by Bacillus sp. in the fermented meju. The use of glasswort and rice in the meju-making process did not alter the bacterial community responsible for the fermentation of meju.     

Identification of a new phenolic compound from Chinese olive (<Emphasis Type="Italic">Canarium album</Emphasis> L.) fruit

Chinese olive (Canarium album L.), a native and a well-known tropical fruit tree in the southeast of China, contains large amount of phenolics and possesses great pharmacological activities. In this study, phenolics were extracted from Chinese olive fruit pulp using 80% (v/v) aqueous acetone, and acetone extracts were further fractioned with petroleum ether, ethyl acetate and n-butanol sequentially. From n-butanol fraction, a new phenolic compound was isolated and purified through AB-8 adsorption resin column chromatography, polyamide column chromatography and TSK Toyopearl HW-40 (S) column chromatography, and the structure of the new compound was established as 3-O-galloyl quinic acid butyl ester by electrospray ionization mass spectrometry (ESI-MS), 1D- and 2D-NMR (DEPT, COSY, HMBC, HMQC) and UV–vis techniques.     

Estrogenic effects of various extracts from <Emphasis Type="Italic">Chamdanggui</Emphasis> (<Emphasis Type="Italic">Angelica gigas</Emphasis> Nakai) and <Emphasis Type="Italic">sogdan</Emphasis> (<Emphasis Type="Italic">Phlomis umbrosa</Emphasis> Turcz)

The various extracts from chamdanggui (Angelica gigas Nakai) and sogdan (Phlomis umbrosa Turcz) were evaluated for estrogenic activity and characterized according to HPLC profile. Chamdanggui and sogdan were individually extracted with 4 solvents (hot water, 70% ethanol, n-butanol, and dichloromethane) of differing polarities. Estrogenic activity was determined by E-screen using an estrogen-dependent MCF-7 BUS cell. Although almost all extracts showed estrogenic effects in a concentrationdependent manner, the hot water extract from chamdanggui (250 μg/mL) had the higher effect (138%). Among 90 fractions using HPLC separation of the hot water extract from chamdanggui, fraction 21 and 28 produced the highest estrogenic effects of 178 and 163% at 10 μg/mL, respectively. The results imply that the hot water extract from chamdanggui could be useful as an alternative hormone replacement therapy.     

Quantification of DNA during winemaking by fluorimetry and <Emphasis Type="Italic">Vitis vinifera</Emphasis> L.-specific quantitative PCR

The ability to genotype wine would be of a great benefit to control the varietal wine authenticity. In this paper, we assessed the fate of grapevine DNA and total DNA during vinification to suggest the latest sampling point that would allow genotyping. The total DNA concentration was measured by fluorimetry and the specific Vitis vinifera L. DNA concentration was measured by quantitative PCR in samples obtained from different steps of vinifications of four wines. To elucidate the effect of pre-extraction and extraction steps on DNA yield from wine, several samples from bottle of wine, two precipitation salts, and two DNA extraction solutions were tested. It was found that hundreds of genome equivalents of grapevine DNA can be extracted from every winemaking sample and from wine. In tested samples, differences in DNA yield among wines from different grapevine varieties, differences among extraction methods, and sampling variation in wine were not statistically significant.     

Antihyperglycemic activity of polyphenolic components of black/bitter cumin <Emphasis Type="Italic">Centratherum anthelminticum</Emphasis> (L.) Kuntze seeds

The effect of black/bitter cumin seeds Centratherum anthelminticum (L.) Kuntze extract (CA) containing mixture of polyphenolic compounds was tested on rat intestinal α-glucosidases, human salivary α-amylase activity and postprandial hyperglycemia in rats. Polyphenolic components of C. anthelminticum seeds (CA) dose dependently inhibited rat intestinal disaccharidases. IC₅₀ values were found to be 34.1 ± 3.8, 62.2 ± 4.5 and 500.5 ± 11.9 μg of CA for rat intestinal sucrase, maltase and p-nitrophenyl α-d-glucopyranoside (PNP-glycoside), respectively. CA also inhibited human salivary α-amylase activity with IC₅₀ value of 185.5 ± 4.9 μg. The inhibitory effect of CA was found to be 8–32 fold more potent than dl-catechin but less effective than acarbose on rat intestinal disaccharidases and salivary α-amylase. The enzyme kinetic studies showed a non-competitive type of inhibition with a low K _i value of 30.24 μg, 76.67 μg and 341.60 μg of CA for maltase, sucrase and PNP-glycoside hydrolysis activities, respectively. The in vitro inhibition of glucosidases was further confirmed by in vivo maltose tolerance test in rats. Feeding of CA at 50–200 mg/kg body weight (b.wt) to maltose (2.0 g/kg b.wt), loaded rats significantly reduced the postprandial plasma glucose levels compared with acarbose. The inhibitory components of CA were identified as a mixture of polyphenolic compounds viz., gallic acid, protocatechuic acid, caffeic acid, ellagic acid, ferulic acid, quercetin and kaempferol. This study demonstrated that CA exerts antihyperglycemic effect by decreasing postprandial glucose in rats by modulating α- amylase and glucosidases (sucrase and maltase) activity and thus may be useful for the management of diabetes mellitus.     

Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

Polyphenol composition of peel and pulp of two Italian fresh fig fruits cultivars (<Emphasis Type="Italic">Ficus carica</Emphasis> L.)

An analytical study was carried out on phenols in two fig cultivars (Ficus carica L.), one black and one green. Fresh fruits were peeled and phenols were extracted separately from the peel and the pulp. The extracts were subjected to HPLC-DAD analysis, by monitoring at four wavelengths, 280, 316, 365 and 520 nm for catechins and benzoic acids, hydroxycinnamic acids, flavonols and anthocyanins, respectively. Results showed that phenols are concentrated almost exclusively in the peel, with the black cultivar having the highest content. In particular, the peel was rich in rutin, with amounts from 527 to 1,071 mg/kg (fresh basis) for green and black figs, respectively. The peel of black figs showed an appreciable content of cyanidin 3-O-rutinoside. The pulp of both green and black cultivars contained only cyanidin 3-O-rutinoside, while the cyanidin 3-O-glucoside was found only in the peel of the black figs. Chlorogenic acid and another cinnamic acid were detected in the peel of both cultivars, while benzoic acids and catechins were not present. Alessandra Del Caro and Antonio Piga contributed equally to this study.     

Differences in morphology and composition of skin and pulp cell walls from grapes (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.): technological implications

The morphology and composition of skin and pulp cell walls of four premium red wine grape (Vitis vinifera L.) varieties were studied to determine whether the technological differences observed in previous studies between the four different varieties during the winemaking process (especially, the anthocyanin extractability) could be partly explained by their respective cell wall compositions. Monastrell grapes showed the highest amount of cell wall material in their skins (55–57 mg cell wall/g skin), while Syrah showed the largest amount of cell wall material in the pulp (5.1 mg cell wall/g pulp). The highest percentage of galactose and cellulosic glucose in skin and pulp cell walls was also found in Monastrell grapes. The cell wall composition of Monastrell grapes, together with their morphology, suggests that the firmer pulp and skin are responsible for the difficulties this variety shows for anthocyanin extraction (extractability index varying from 37 to 60). As cell wall composition may be modulated by the action of enzymes, the activity of pectinmethylesterase, polygalacturonase, cellulase and α- and β-galactosidase was studied. No polygalacturonase and cellulase activity was found. The two galactosidases showed higher activity than pectinmethylesterase (25–290 units/g tissue versus 0.3–0.4 units/g tissue), especially in Cabernet Sauvignon grapes, although in Monastrell grapes α- and β-galactosidase activity was low, which may explain the large amount of galactose in Monastrell grapes cell wall.     

Characterisation of certain major polyphenolic antioxidants in grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis> cv. Roditis) stems by liquid chromatography-mass spectrometry

The stem extract, which was shown to contain high amounts of phenolics and significant antioxidant potency, was further analysed employing liquid chromatography-electrospray ionisation mass spectrometry, in an effort to obtain a deeper insight into its polyphenolic composition. The compounds identified were mainly flavanols (a trimer and a galloylated dimer), flavonols (rutin and quercetin 3-O-glucuronide), stilbenes (trans-resveratrol and a resveratrol dehydrodimer) and a dihydroflavonol glycoside (astilbin).     

Biochemical characterization of esterase from soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L.)

Esterases are enzymes that present good potential in industrial application, and soybean seed can represent an alternative source for this enzyme. The extraction and esterase activity of Brazilian soybean seeds (Glycine max L.) were evaluated. Esterase activity was found in both the germinated and non-germinated seeds at 2.90 and 1.70 U/mg, respectively, with a concentration in the powdered extract (freeze dried) of 100 mg/mL. The enzyme showed a preference for the hydrolysis of short chain fatty acids (120.02 U/mL) and optimum pH for activity was pH 8.0 with optimum temperatures of 40 and 80°C. The enzyme showed stability at 70°C showing 60% of residual activity and activity increased with the addition of the following salts: NaNO₃, K₂SO₄, and Na₂SO₄ in the reaction medium.