Application of Ionic Liquid-Based Ultrasound-Assisted Extraction of Five Phenolic Compounds from Fig (<Emphasis Type="Italic">Ficus carica</Emphasis> L.) for HPLC-UV

Ionic liquid-based ultrasonic-assisted extraction (IL-UAE) was successfully developed to extract the known phenolics gallic acid, chlorogenic acid, rutin, psoralen, and bergapten present in the leaves, pulps, and peels of Ficus carica L. In this work, the proposed method was evaluated in comparison with regular ultrasonic-assisted extraction in which methanol was the extractant and exhibited higher efficiency. Moreover, ionic liquids with different cations were investigated and 1-butyl-3-methylimidazole hexafluorophosphate solution was selected as the optimal extractant at concentration of 1.0 mol L^?1. The corresponding extraction parameters including extraction solvent, the concentration of [BMIM][PF₆], solid–liquid ratio (g mL^?1), ultrasonic extraction time, and extraction temperature were optimized. The phenolics were then determined by high-performance liquid chromatography (HPLC). Under the optimized conditions, the content of gallic acid, chlorogenic acid, rutin, psoralen, and bergapten in leaves, pulps, and peels of F. carica L. was 12.67–37.16, 14.75–90.07, 107.91–222.37, 2.59–67.83, and 1.99–20.21 μg/0.1 g, respectively. This study suggests that IL-UAE is a rapid, simple, and green preparation technique.     

Process Intensification by Experimental Design Application to Microwave-Assisted Extraction of Phenolic Compounds from <Emphasis Type="Italic">Juglans regia</Emphasis> L.

Microwave-assisted extraction was applied to Juglans regia L. fresh male flowers and unripe walnut seeds to evaluate the total phenolic contents of the extracts as well as the percentage of water-soluble polyphenols. The research was planned using the Design of Experiments technique to investigate the role on the extraction efficiency of different parameters, such as temperature, time and number of microwave heating cycles, together with their possible interactions. Optimization was achieved by applying a Response Surface Methodology comprising a three-factor, two-level, full-factorial Face-Centred Central Composite Design. The two input variables with a significant effect on the recovery of phenols from fresh male flowers were the extraction temperature and the number of microwave cycles. In the case of unripe walnut seeds, a linear two-factor interaction model was selected, with significant interactions occurring between temperature and time, and time and number of microwave cycles. The best experimental conditions were as follows: 100 °C, 6 min, three microwave heating cycles and 22.7 ± 0.2 mg gallic acid equivalent (GAE)/g (total phenolic content, PC); energy consumption calculations suggested slightly different conditions: 60 °C, 30 min, three microwave heating cycles and 20.7 ± 0.3 mg GAE/g (TPC).     

Antioxidant and tyrosinase inhibitory activities of different parts of guava (<Emphasis Type="Italic">Psidium guajava</Emphasis> L.)

The antioxidant and the tyrosinase inhibitory activities of 4 different solvents (acetone, ethanol, methanol, and water) for preparation of extracts from guava (branch, fruit, leaf, and seed) were evaluated by measuring total phenolic contents (TPC), DPPH radical scavenging activity, ABTS radical scavenging activity, reducing power (RP), and tyrosinase inhibitory activity. The extracts of branch and leaf showed relatively higher antioxidant properties than those of fruit and seed. The highest TPC (141.28 mg/g gallic acid equivalents), DPPH radical scavenging activity (IC₅₀=34.01 μg/mL), ABTS radical scavenging activity (IC₅₀=3.23 μg/mL), and RP (IC₅₀= 75.63 μg/mL) were found in acetone extract of leaf, while water extract of seed had the lowest antioxidant activity. The tyrosinase inhibitory activity of ethanol extract from guava leaf was 69.56%, which was the highest activity among the extracts. These results indicate that useful bioactive substances exist in the guava branch as well as leaf extracts.     

Non-destructive Assessment of Guava (<Emphasis Type="Italic">Psidium guajava</Emphasis> L.) Maturity and Firmness Based on Mechanical Vibration Response

Storage potential and eating quality of guava (Psidium guajava L.) fruit depend on its maturity. Segregation of guava according to maturity and firmness measured using non-destructive technologies would help the industry to designate ripe fruit to immediate market and less ripe fruit for distant market (e.g., exportation). This research was conducted to evaluate the potential of experimental resonant frequency (f _e) and elasticity index (EI) to estimate fruit firmness, which has been reported to be inversely correlated to its maturity. A maturity index (I _m) was calculated as the ratio of total soluble solids/titratable acidity (TSS/TA). It was proved that TSS, TA, and I _m were significantly correlated (P?<?0.05) to skin firmness (F _s), flesh firmness (F _f), stiffness (S), and analytical resonant frequency (ω _n ), being S the attribute best fitted to I _m (R ²?=?0.77). Since it was observed that f _e and EI were sensitive to changes in fruit firmness, both of them were explored as alternatives to predict F _s, F _f, S, and ω _n of guava fruit. In some cases, EI improved the models to predict guava firmness traits (e.g., F _s vs f _e had a coefficient of determination of R ²?=?0.58, whereas for F _s vs EI, it was R ²?=?0.62). The best model occurred when plotting ω _n vs f _e (R ²?=?0.86), followed by S vs EI (R ²?=?0.84), making these promising features for the development of a new practical application using frequency response measurement as a non-destructive method to assess guava maturity.     

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.     

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.     

Phenolic contents and antioxidant activities from different tissues of <Emphasis Type="Italic">Baekseohyang</Emphasis> (<Emphasis Type="Italic">Daphne kiusiana</Emphasis>)

The aim of this research was to investigate the levels of phenolic compounds and antioxidant activities in different tissues including leaves, stems, and roots from baekseohyang (Daphne kiusiana). The highest contents of total phenolics (43.59 mg gallic acid equivalent, GAE/g) and flavonoids (15.73 mg rutin equivalents, RE/g) were observed in the 75% methanol extract of leaves. Moreover, this extract had the predominant antioxidant capacity, DPPH (85.91%) and ABTS (92.57%) radical scavenging activities as well as reducing power (7.20%) at a concentration of 5 mg/mL. The highest content of phenolic compounds was also exhibited in this extract with an increasing order in leaves, roots, and stems and their major components were vanillic acid (6.37 mg/g), tannic acid (1.91 mg/g), and p-hydroxybenzoic acid (3.96 mg/g). Thus, the strong antioxidant activities of the 75% methanol extract are correlated with high phenolic compound contents. This study suggests that baekseohyang leaves may potentially be used as an accessible source of natural antioxidants.     

A New Multiplexed Real-Time PCR Assay to Detect <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>, <Emphasis Type="Italic">C. coli</Emphasis>, <Emphasis Type="Italic">C. lari</Emphasis>, and <Emphasis Type="Italic">C. upsaliensis</Emphasis>

A new rapid method based on real-time PCR was developed to detect four thermophilic Campylobacter species (Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis) in food samples. The assay targeted the bipA gene for C. upsaliensis and C. lari, whereas the gene encoding the ATP-binding protein CJE0832 was used to detect C. coli and C. jejuni. These genes were chosen for this assay due to their low variability and mutation rate at a species level. The multiplex PCR showed 100% inclusivity for all 25 thermophilic Campylobacter strains tested and 100% exclusivity for 38 non-targeted strains belonging to closely related species. The newly developed real-time PCR could detect down to 10² genomes/reaction and displayed efficiency above 97% for all species except for C. upsaliensis (90.1%). The method proved to be a reliable tool for food analysis, showing 100% sensitivity, 96% efficiency, and 92.45% specificity when validated against the gold standard method UNE-EN ISO 10272:2006 using 200 diverse food samples (meat, fish, fruits and vegetables, and raw milk). In artificially spiked samples, the detection limit of the method was 10 cfu/g in salad, 5 cfu/g in turkey meat, and 1 cfu/g in the rest of meat samples tested. Consequently, the newly designed molecular tool represents a quick and safe alternative to obtain reliable results concerning the presence/absence of the main thermophilic Campylobacter in any food sample.     

Thermal inactivation kinetics of quality-related enzymes in cauliflower (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">botrytis</Emphasis><Emphasis Type="Bold">)</Emphasis>

Thermal inactivation of quality-related enzymes in both cauliflower crude enzyme extracts and fresh tissue samples was studied in temperature range 50–100 °C. For crude enzyme extracts, several parameters, reaction rate constants (k) and activation energy (E _a) as well as decimal reduction time (D) and (z) values, were used to characterize the thermal stability. The rates of inactivation were found to follow first-order inactivation kinetics. Activation energies varied between 101.18 and 208.42 kJ mol⁻¹ with z values of 10.59–24.09 °C. The examined kinetics indicated that lipoxygenase was the most heat resistant followed by peroxidase, polyphenol oxidase, pectin methyl esterase and ascorbic acid oxidase. Furthermore, the obtained results from the blanched fresh tissues indicated that inactivation of lipoxygenase secured disappearing of any other enzyme activities. Therefore, this study recommends using lipoxygenase as an indicator enzyme to optimize the thermal treatments of cauliflower products.     

In vivo study of antiallergenicity of ethanol extracts from <Emphasis Type="Italic">Sargassum tenerrimum</Emphasis>, <Emphasis Type="Italic">Sargassum cervicorne</Emphasis> and <Emphasis Type="Italic">Sargassum graminifolium turn</Emphasis>

Food allergy has becoming the serious threat in the world for which the search of an effective anti-allergic drug is the demand of time. Keeping in view of the potentiality of seaweeds, the ethanol extracts from Sargassum tenerrimum (ST), Sargassum cervicorne (SC), and Sargassum graminifolium turn (SG) have been studied in vivo for its antiallergenicity through passive cutaneous anaphylaxis (PCA) and active cutaneous anaphylaxis (ACA) in female BALB/c mice. Intraperitoneal administration of these ethanol extracts inhibit mouse PCA and ACA in a dose-dependent manner using ovalbumin (OVA) and shrimp allergen as triggering agents to induce allergenicity over mice. The extract of ST containing phlorotannin has been found most active over the suppression of PCA triggered by OVA and shrimp with IC₅₀ values of 25.64 and 40.98 mg/kg, respectively and an efficacy comparable to that of an anti-allergic drug disodiumcromoglycate. Similarly, ST inhibits ACA triggered by ova and shrimp allergen in the mouse, with 50% suppression at 25.5 and 43.53 mg/kg, respectively. The results presented here show that these extracts are active on the studied models among which ethanol extract of ST was the most potent, leading toward the promising development of a new class of anti-allergic drugs.     

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.     

Molecular diversity among natural populations of <Emphasis Type="Italic">Lactobacillus paracasei</Emphasis> and <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis>/<Emphasis Type="Italic">paraplantarum</Emphasis> strains isolated from autochthonous dairy products

The diversity of 87 Lactobacillus paracasei and Lactobacillus plantarum/paraplantarum strains, previously identified from different autochthonous dairy products, was investigated by phenotypic and genotypic approaches. The increased resolution obtained using phenotypic and genotypic characterization allowed the level of strain heterogeneity detection to be widened. Phenotypic diversity was evaluated by studying biochemical characteristics of technological interest, including antimicrobial and proteinase activities, resistance to nisin, aggregation ability, production of exopolysaccharides, acetoin and diacetyl, citrate utilization, and antibiotic susceptibility. Genotypic diversity was generally evaluated by PCR amplification of repetitive bacterial DNA element fingerprinting using the (GTG)₅ primer [(GTG)₅-PCR]. Moreover, in cases where strains were not discriminated by (GTG)₅-PCR combined with phenotypic analysis, pulsed-field gel electrophoresis (PFGE) analysis was performed. The results indicate that L. plantarum/paraplantarum and L. paracasei natural isolates from artisanal dairy products are a gold mine in terms of diversity of strains and could be potentially interesting to dairy companies for the formulation of functional starter cultures in the production of innovative foods.     

Alternative Extraction Method of Bioactive Compounds from Mulberry (<Emphasis Type="Italic">Morus nigra</Emphasis> L.) Pulp Using Pressurized-Liquid Extraction

Black mulberry (Morus nigra L.) is one of the most important species of the genus Morus as its fruit contains substantial levels of anthocyanins and phenolic compounds which show a potentially positive effect on the human health. Nowadays, PLE is becoming a promising extraction technology. Therefore, the development of fast extraction methods of anthocyanins and phenolic compounds from mulberry pulp using pressurized-liquid extraction (PLE) has been studied in this paper. The operating conditions (solvent, temperature, pressure, purge time, pH, and flushing) were investigated by a Box–Behnken design. Analysis of the model clearly showed that the most influential factors were temperature and solvent composition. The optimum extraction conditions for anthocyanins were 47.2% methanol in water, a temperature of 75.5 °C, pressure of 200 atm, a purge time of 90 s, pH 3.01, and 50.2% for flushing. The best conditions for the extraction of phenolics were 74.6% methanol, 99.4 °C, 100 atm, 90 s purge, pH 7, and 100% flushing. The optimum extraction time was 10 min. The precision values of the methods were also evaluated and excellent results (RSD?<?5%) were obtained. The developed methods were successfully applied to several mulberry marmalade samples. The results using PLE were compared to those achieved by UAE methods. Similar extraction yields were obtained for anthocyanins by PLE and UAE under optimized conditions; however, PLE required less methanol consumption. Besides, PLE showed higher extraction efficiency for total phenolic compounds. From the results, it can be concluded that pressurized-liquid extraction can be considered as an efficient alternative and powerful tool for the extraction of bioactive compounds from mulberries.     

Prevalence,genetic diversity,and antibiotic susceptibility of <Emphasis Type="Italic">Cronobacter</Emphasis> spp. (<Emphasis Type="Italic">Enterobacter sakazakii</Emphasis>) isolated from <Emphasis Type="Italic">Sunshik</Emphasis>, its ingredients and soils

The prevalence, genetic diversity, and antibiotic susceptibility of Cronobacter species (Enterobacter sakazakii) isolated from sunshik products, its ingredients, and root vegetable farm’s soils were investigated to analyze main reservoirs and contaminated sources of Cronobacter spp. Cronobacter spp. was isolated from 9 of 15 sunshik products, 26 of 72 its ingredients, and 2 of 39 soils. The root vegetables such as sweet potato and carrot showed higher contamination rate (70%) than the other sunshik ingredients. All isolates showed 929 bp band amplified from 16S rRNA and resistant to ampicillin, amoxicillin/clavulanic acid, and cefazolin. All isolates showed diverse random-amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) band patterns. However, 3 cases of RAPD banding patterns between clinical strains and isolates from sunshik products and root vegetables (yam, carrot) were related with similarities level of 80%. These studies indicated that root vegetables can be an important contamination source of Cronobacter spp. in sunshik products. Thus, the preparation of root vegetables for manufacturing sunshik products used as a weaning diet was handled with more care than the other sunshik ingredients.     

Effect of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> Bauer and <Emphasis Type="Italic">Bifidobacterium animalis</Emphasis> ssp. <Emphasis Type="Italic">lactis</Emphasis> BB12 on proteolytic changes in dry-cured loins

The effect of the potentially probiotic bacteria strain of Lactobacillus acidophilus Bauer and probiotic bacteria Bifidobacterium animalis ssp. lactis BB12 on proteolytic changes of proteins in dry-cured loins during fermentation and cold storage was studied. Results of the conducted tests demonstrated that the use of probiotic bacteria for the production of dry-cured meats impacts the generation of products of protein proteolysis with high antioxidant activity. The highest antioxidant activity of peptides after fermentation and cold storage was observed in the loin with the strain B. animalis ssp. lactis BB12 and the loin with the mixture of strains L. acidophilus Bauer and B. animalis ssp. lactis BB12. Qualitative analysis of peptides demonstrated that peptides with weight below 3.5 kDa are characterized by the highest capacity of quenching the ABTS cation radical, including the peptides in loins with the strain B. animalis ssp. lactis BB12.     

Characterization of brewing microorganisms isolated from Korean traditional <Emphasis Type="Italic">nuruk</Emphasis> for <Emphasis Type="Italic">Cheongju</Emphasis> production

The objective of this study was to select and identify yeasts and molds isolated from traditional nuruk and to investigate their brewing characteristics for Cheongju production. The yeast strains Y190 (Accession ID-KACC 93251P), Y263 (Accession ID-KACC 93252P), and Y270 (Accession ID-KACC 93253P) showing high alcohol and flavor productivity were isolated and identified by phylogenetic inference based on an internal transcribed spacer 2 region sequence analysis. In addition, Aspergillus oryzae 83-10 (Accession ID-KACC 93254P) showing the highest enzyme activity was isolated. This study provides basic data for the production of Korean Cheongju and assesses the applicability of these three yeast strains and A. oryzae 83-10 isolated from traditional nuruk.     

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.     

Optimization of Conditions for the Extraction of Antioxidants from Leaves of <Emphasis Type="Italic">Syzygium cumini</Emphasis> L. Using Different Solvents

The aim of the present study was to find out the best method for extracting antioxidants from Syzygium cumini L. leaves. The extraction was done by three different methods: sequential cold percolation extraction method, decoction extraction method, and maceration extraction method. Antioxidant activity, total phenol, and flavonoid content were determined in all different extracts of various extraction methods of S. cumini L. leaves. Antioxidant activity was tested by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical, superoxide anion radical and hydroxyl radical scavenging activities, and reducing capacity assessment. Sequential cold percolation extraction method proved to be the best extraction method. The acetone extract had maximum phenol and flavonoid content and showed best DPPH free radical scavenging activity and reducing capacity assessment. Ethyl acetate extract showed best superoxide radical scavenging activity, while aqueous extract showed best hydroxyl radical scavenging activity. It can be concluded that sequential cold percolation extraction method is the best method of extracting leaf antioxidants for this plant at least.     

Ethanolic fermentation from red ginseng extract using <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> and <Emphasis Type="Italic">Saccharomyces carlsbergensis</Emphasis>

The pH of red ginseng extracts fermented with Saccharomyces cerevisiae and Saccharomyces carlsbergensis decreased rapidly during 3 days of fermentation, with no further significant change thereafter. After 20 days of fermentation, a relatively small difference remained in the acidity of extracts fermented with S. cerevisiae (0.54%) and S. carlsbergensis (0.58%). Reducing sugar in the S. cerevisiae and S. carlsbergensis extracts decreased from 258.6 to 45.4 and 43.2 mg/mL glucose equivalents, respectively; and ethanol contents increased from 1.5% at day 0 to 16.0 and 15.0%, respectively, at day 20. Ginsenosides Rb₁, Rb₂, Rc, Re, Rf, and Rg₁ decreased during the fermentation with S. cerevisiae, but Rd and Rg₃ increased by day 12. Ginsenosides Rb₁, Rb₂, Rc, Re, and Rg₁ decreased gradually in the extract with S. carlsbergensis, but Rd and Rg₃ were increased at day 6 and 9, respectively.