In vitro inhibitory effects of cranberry bean (Phaseolus vulgaris L.) extracts on aldose reductase, α‐glucosidase and α‐amylase

The in vitro inhibitory activities of different seed extracts prepared from cranberry bean mutant SA‐05 and its wild‐type variety Hwachia against aldose reductase, α‐glucosidase and α‐amylase were examined. The results indicated that the polyphenolics‐rich extracts obtained using 800 g kg^?1 methanol and 500 g kg^?1 ethanol demonstrated inhibitory activities against aldose reductase (IC₅₀ of 0.36–0.46 mg mL^?1) and α‐glucosidase (IC₅₀ of 1.32–1.94 mg mL^?1). The 500 g kg^?1 ethanol extracts also showed α‐amylase inhibitory activities (IC₅₀ of 70.11–80.22 μg mL^?1). Subsequent extracts, prepared further with NaCl and H₂O from precipitates of 800 g kg^?1 methanol or 500 g kg^?1 ethanol extracts, exhibited potent α‐amylase inhibitory activities (IC₅₀ of 17.68–38.68 μg mL^?1). A combination of 500 g kg^?1 ethanol extraction plus a subsequent H₂O extraction produced highest polyphenolics and α‐amylase inhibitors. The SA‐05 α‐amylase inhibitor extracts showed greater inhibitory activities than that of Hwachia. Thus, cranberry bean mutant SA‐05 is an advantageous choice for producing anti‐hyperglycaemic compounds.     

The influence of heat treatment of chickpea seeds on antioxidant and fibroblast growth‐stimulating activity of peptide fractions obtained from proteins digested under simulated gastrointestinal conditions

This study presents the effect of heat treatment of chickpea seeds on biological activity of peptides obtained by in vitro gastrointestinal digestion. The most significant antiradical activity against ABTS^+? expressed as IC₅₀ value was observed for 3.5‐ to 7‐kDa peptide fraction from TC hydrolysate (41.01 μg mL^?1). In turn, peptide fraction of 3.5–7.0 kDa obtained from raw chickpea seeds hydrolysate showed the highest antiradical activity against DPPH^? and Fe²⁺ chelating activity with IC₅₀ value of 20.94 and 52.53 μg mL^?1, respectively. The highest Cu²⁺ chelating activity was observed for peptides obtained from TC hydrolysate (IC₅₀ = 56.60 μg mL^?1). Peptide fraction <3.5 kDa from TC hydrolysate demonstrated the most significant reducing power (0.362 A₇₀₀/μg mL^?1). The peptide fraction of 3.5–7 kDa from TC hydrolysate also showed the highest fibroblast growth‐stimulating activity. These results indicated that the heat treatment process has no significant effect on antiradical activity against DPPH^? and Fe²⁺ chelating ability of peptides.     

Phytosterols in banana (Musa spp.) flower inhibit α‐glucosidase and α‐amylase hydrolysations and glycation reaction

Three phytosterols were isolated from Musa spp. flowers for evaluating their capabilities in inhibiting glucosidase and amylase activities and glycation of protein and sugar. The three phytosterols were identified as β‐sitosterol (PS1), 31‐norcyclolaudenone (PS2) and (24R)‐4α, 14α, 4‐trimethyl‐5α‐cholesta‐8, 25(27)‐dien‐3β‐ol (PS3). IC₅₀ values (the concentration of inhibiting 50% of enzyme activity) of PS1, PS2 and PS3 against α‐glucosidase were 283.67, 11.33 and 43.10 μg mL^?1, respectively. For inhibition of α‐amylase, the IC₅₀ values of PS1, PS2 and PS3 were 52.55, 76.25 and 532.02 μg mL^?1, respectively. PS1 was an uncompetitive inhibitor against α‐amylase with K_m at 5.51 μg mL^?1, while PS2 and PS3 exhibited a mixed‐type inhibition with K_m at 52.36 and 2.49 μg mL^?1, respectively. PS1 and PS2 also significantly inhibited the formation of advanced glycation end products (AGEs) in a BSA–fructose model. The results suggest that banana flower could possess the capability in prevention of the diseases associated with abnormal blood sugar and AGEs levels, such as diabetes.     

Chemical composition,antioxidant activity and anti‐lipase activity of Origanum vulgare and Lippia turbinata essential oils

This study reported the chemical composition, phenolic content, antioxidant and anti‐lipase activity of oregano and Lippia essential oils. The major compounds found in oregano essential oil were γ‐terpinene (32.10%), α‐terpinene (15.10%), p‐cymene (8.00%) and thymol (8.00%). In Lippia essential oil, α‐limonene (76.80%) and 1,8‐cineole (4.95%) represented the major compounds. Oregano essential oil had higher phenolic content (12.47 mg gallic acid mL^?1) and DPPH scavenging activity (IC₅₀ 0.357 μg mL^?1) than Lippia essential oil (7.94 mg gallic acid mL^?1 and IC₅₀ 0.400 μg mL^?1, respectively). Both essential oils had similar antioxidant indexes (about 1.2) determined by Rancimat. Moreover, oregano essential oil had also higher anti‐lipase activity (IC₅₀ 5.09 and 7.26 μg mL^?1). Higher phenolic content in the essential oils was related with higher scavenging and anti‐lipase activities. Oregano and Lippia essential oils could be used as natural antioxidants on food products.     

Tannin fraction from Ampelopsis grossedentata leaves tea (Tengcha) as an antioxidant and α‐glucosidase inhibitory nutraceutical

Leaf of Ampelopsis grossedentata is a new resource of functional foods with healthful properties. Antioxidant and α‐glucosidase inhibitory activities of water extract (made in the style of drinking), tannin fraction (TF) and dihydromyricetin (DMY) from A. grossedentata leaves were evaluated. The main component of TF was identified as gallotannins. DPPH and ABTS radical scavenging activities and reducing power of TF were superior to those of water extract, however, inferior to those of DMY. In no PBS wash protocol of cellular antioxidant activity assay, DMY and TF exhibited similarly, while in PBS wash protocol, the value of TF was higher than that of DMY. In addition, TF possessed the highest α‐glucosidase inhibitory activities (IC₅₀ = 1.94 μg mL^?1), followed by water extract (IC₅₀ = 23.10 μg mL^?1) and DMY (IC₅₀ = 72.21 μg mL^?1). The strong α‐glucosidase inhibitory activity of TF may attribute to the binding capacity to enzymes, as confirmed by fluorescence analysis.     

Phytochemicals content,antioxidant and hypoglycaemic activities of commercial nutmeg mace (Myristica fragrans L.) and pimento (Pimenta dioica (L.) Merr.)

Commercially dried powder of nutmeg mace (Myristica fragrans) and pimento (Pimenta dioica) spices was investigated for their high performance liquid chromatography phenolic profile and their antioxidant and hypoglycaemic properties by α‐amylase and α‐glucosidase inhibition tests. Generally, mace showed the most promising activity. An interesting protection of lipid peroxidation with an IC₅₀ value of 7.7 μg mL^?1 was found. A significant result was also obtained in ferric reducing ability power assay if compared to the positive control butylated hydroxytoluene (IC₅₀ value of 68.7 μg mL^?1 vs. 63.2 μg mL^?1, respectively). Mace also exhibited the highest carbohydrate‐hydrolysing enzyme inhibitory activity with IC₅₀ values of 62.1 and 75.7 μg mL^?1 against α‐amylase and α‐glucosidase, respectively. Overall, these results support the use of these spices not only as flavouring agent but also as food preservative and functional ingredients.     

Effects of physicochemical factors and in vitro gastrointestinal digestion on antioxidant activity of R‐phycoerythrin from red algae Bangia fusco‐purpurea

R‐phycoerythrin (R‐PE) was purified from the red algae Bangia fusco‐purpurea after 35–50% ammonium sulphate fractionation followed by ion‐exchange column chromatography on DEAE‐Sepharose, resulting in a purity (A₅₆₅/A₂₈₀) ratio of 5.1. The circular dichroism spectroscopy results suggested that the structure of R‐PE is predominately helical. The antioxidant activity of R‐PE was studied and revealed changes in conformation and antioxidant activity at different temperatures and pH values. After in vitro‐simulated gastrointestinal (GI) digestion of R‐PE, the scavenging activity of ABTS radical (EC₅₀, 769.9 μg mL^?1), DPPH radical (EC₅₀, 421.9 μg mL^?1), hydroxyl radical (EC₅₀, 32.4 μg mL^?1) and reducing power (A₇₀₀ = 0.5, 625.8 μg mL^?1) were measured. Gel filtration chromatography analysis showed that the molecular weight distribution of the final GI digest that still contained high antioxidant activity was <3 kDa. Our present results indicate that digestion‐resistant antioxidant peptides of R‐PE may be obtained by in vitro GI proteinases degradation.     

Inhibitory activity towards human α‐amylase in wheat flour and gluten

Wheat α‐amylase inhibitors (AI) have been targeted as potential triggers of noncoeliac gluten or wheat sensitivity (NCGS). The aim of this study was to determine AI activity towards α‐amylase from human source in wheat cultivars. Contrary to barley, buckwheat, or oats, high level of AI activity was found in wheat and, to a lesser extent, rye. AI activity (mean IC₅₀ = 137 μg mL^?1) did not vary with respect to ancient or recently developed wheat cultivars. Vital wheat gluten had very high and heat‐stable AI activity (mean IC₅₀ = 23 μg mL^?1), higher than wheat starch (?10 000 μg mL^?1) or acarbose (40 μg mL^?1), a medication for the management of hyperglycaemia and potentially causing digestive disorders due to the accumulation of undigested carbohydrates in the intestine. Data suggest that eating raw wheat gluten, flour or dough could pose a health risk.     

Comparative study of the essential oils of three Beilschmiedia species and their biological activities

This study was designed to examine the chemical compositions of the essential oils from three Beilschmiedia species and antioxidant, antimicrobial, antityrosinase, acetylcholinesterase and anti‐inflammatory activities. The essential oils of B. kunstleri, B. maingayi, B. penangiana gave β‐caryophyllene (10.6–12.1%), β‐eudesmol (17.5–24.1%) and δ‐cadinene (17.5–28.7%) as the most abundant components respectively. The bark oil of B. maingayi showed the highest activity in β‐carotene/linoleic acid (125.9%) and phenolic content (288.2 mg GA g^?1), while B. penangiana bark oil was found to have strong activity in DPPH (IC₅₀ 84.7 μg mL^?1) and ABTS (IC₅₀ 108.3 μg mL^?1). The essential oils of B. penangiana showed the best activity against Candida glabrata with MIC value 31.3 μg mL^?1. The bark oil of B. penangiana gave 82.5% tyrosinase inhibiton. The leaf oil of B. maingayi gave the highest inhibition in AChE (66.6%) and lipoxygenase (77.0%) assay. Our findings demonstrate that the essential oils have great potential for applications in pharmaceutical and food industries.     

Separation of angiotensin I‐converting enzyme inhibitory peptides from bovine connective tissue and their stability towards temperature,pH and digestive enzymes

Bovine collagen was isolated from connective tissue, a by‐product in the meat processing industry and characterised by SDS‐PAGE. Alcalase and papain were employed to generate collagen hydrolysates with different degree of hydrolysis (DH). In vitro angiotensin I‐converting enzyme (ACE) inhibitory activities were evaluated and the two most potent hydrolysates from each enzyme were separated by two‐step purification. Both alcalase‐catalysed and papain‐catalysed hydrolysates exhibited strong ACE inhibitory capacities with IC₅₀ values of 0.17 and 0.35 mg mL^?1, respectively. Purification by ion‐exchange chromatography and gel filtration chromatography revealed higher ACE inhibitory activities in one fraction from each enzyme with IC₅₀ values of 3.95 and 7.29 μg mL^?1. These peptide fractions were characterised as 6‐12 amino acid residues by MALDI‐TOF/MS. The peptides retained their activity (>90%) after exposure to processing temperature and pH and in vitro simulated gastrointestinal digestion. The present results demonstrated that collagen peptides can be utilised for developing high value‐added ingredients, for example ACE inhibitory peptides.     

Angiotensin I‐converting enzyme inhibitory peptides FQPSF and LKYPI identified in Bacillus subtilis A26 hydrolysate of thornback ray muscle

Angiotensin I‐converting enzyme (ACE) inhibitory peptides have been searched in thornback ray (Raja clavata) muscle hydrolysed with Bacillus subtilis A26 proteases until a hydrolysis degree of 18.35%. The hydrolysate showed an IC₅₀ of 0.83 mg mL^?1. To identify peptides responsible for this activity, the extract was eluted through size‐exclusion chromatography and fractions collected. The highest ACE inhibitory activity was found for fractions F2 and F3 which had IC₅₀ of 0.42 and 0.51 mg mL^?1, respectively. These fractions were analysed by nano‐liquid chromatography coupled to tandem mass spectrometry (nLC‐MS/MS). A total of 131 and 108 peptide sequences mainly derived from actin, myosin heavy chain and procollagen alpha 1 chain proteins were identified in fractions F2 and F3, respectively. FQPSF and LKYPI showed the best results with an IC₅₀ of 12.56 and 27.07 μM, respectively. These results prove the potential of thornback ray muscle hydrolysate as a source of ACE inhibitory peptides.     

Development of polyclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for sodium saccharin residue in food samples

A sensitive and specific polyclonal antibody (PcAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) for sodium saccharin is described. 6-Amino saccharin was coupled to carrier protein for artificial antigen by diazotisation. New Zealand white rabbits were immunised to obtain anti-sodium saccharin PcAb and then icELISA was developed. The assay showed high sensitivity and specificity to sodium saccharin, with the 50% inhibition value (IC₅₀) of 0.243 μg mL⁻¹, workable range (IC₃₀–IC₇₀) of 0.050–12.8 μg mL⁻¹ and limit of detection (LOD, IC₂₀) of 0.021 μg mL⁻¹. The average recoveries of sodium saccharin in spiked food samples were estimated ranging from 70.7% to 98.8%. A statistically significant correlation of results was obtained between this new ELISA and previously established HPLC approaches with the food-relevant sodium saccharin concentration range 0–320 μg mL⁻¹ (R² = 0.9887–0.9975). These results indicated that the established ELISA was a potential and useful analytical tool for rapid determination of sodium saccharin residue in food samples.     

Effects of fermentation conditions on the potential anti‐hypertensive peptides released from yogurt fermented by Lactobacillus helveticus and Flavourzyme®

This study investigates the effects of fermentation conditions on the production of angiotensin‐converting enzyme inhibitory (ACE‐I) peptides in yogurt by Lactobacillus helveticus 881315 (L. helveticus) in the presence or absence of Flavourzyme^®, which is derived from a mould, Aspergillus oryzae and used for protein hydrolysis in various industrial applications. Optimal conditions for peptides with the highest ACE‐I activity were 4% (v/w) inoculum size for 8 h without Flavourzyme^® supplementation, and 1% inoculum size for 12 h when combined with Flavourzyme^®. The yogurt fermented by L. helveticus resulted in IC₅₀ values (concentration of inhibitor required to inhibit 50% of ACE activity under the assayed conditions) of 1.47 ± 0.04 and 16.91 ± 0.25 mg mL^?1 with and without Flavourzyme^® respectively. Seven fractions of ACE‐I peptides from the yogurt incorporated with L. helveticus and Flavourzyme^® were separated using the preparative high‐performance liquid chromatography. Fraction (F3) showed the highest ACE‐I activity with an IC₅₀ of 35.75 ± 5.48 μg mL^?1. This study indicates that yogurt may be a valuable source of ACE‐I peptides, which may explain the outcomes observed in the experimental and clinical studies and foresee the application of fermented milk proteins into functional foods or dietary supplements.     

A sensitive and selective direct competitive enzyme-linked immunosorbent assay for fast detection of Sudan I in food samples

BACKGROUND: Sudan I, a synthetic azo dye, is considered to be a genotoxic carcinogen and is prohibited in foodstuffs for any purpose at any level worldwide. In this study, a sensitive and specific direct competitive enzyme‐linked immunosorbent assay (dc‐ELISA) for fast detection of Sudan I in food samples was developed for the first time. The monoclonal antibody against Sudan I was used as capture protein, while horseradish peroxidase labeled Sudan I conjugate prepared by the periodate method via ovalbumin (OVA) as a bridge was used as enzyme tracer. RESULTS: The standard curve of dc‐ELISA for Sudan I was constructed in the range 0.1–100 ng mL^?1 and the assay time was within 80 min. Sensitivity was 2.6 ng mL^?1 and the limit of detection was 0.08 ng mL^?1. Cross‐reactivity values of the assay with Sudan II, III and IV were 5.78%, 1.72% and 0.64%; no cross‐reactivity was found with six other edible colorants. The assay was tolerated to 30% of methanol and 10% of acetonitrile without significant loss of IC₅₀. Recoveries of spiked Sudan I in five different samples including chilli powder, tomato sauce, hotpot seasoning and chilli sauce I and II were within 88.4–113.2% and the intra‐assay relative standard deviation was less than 14%. The dc‐ELISA was confirmed by conventional high‐performance liquid chromatography and the correlation coefficient of the two methods was 0.9902. CONCLUSION: The proposed dc‐ELISA method provides an alternative method for sensitive, specific and fast determination of Sudan I in food samples. Copyright © 2011 Society of Chemical Industry     

Polyphenolic composition,antioxidant and antiproliferative effects of wild and cultivated blackberries (Rubus fruticosus L.) pomace

The content of total polyphenolics, antioxidative capacity and antiproliferative activity were tested in wild and cultivated blackberry pomace. Wild blackberry pomace extract Tw2 showed the highest following contents: total polyphenolics (50.16 mg GAE g⁻¹ dw), flavonoids (7.73 mg Qc g⁻¹ dw), flavonols (6.63 mg Qc g⁻¹ dw) and total monomeric anthocyanins (13.40 mg Cy g⁻¹). Tw2 extract significantly inhibited free radicals: IC₅₀^DPPH = 127.76 μg mL⁻¹, IC₅₀^ABTS = 26.53 μg mL⁻¹ and IC₅₀ ^{˙ OH} = 168.62 μg mL⁻¹, and the growth of breast adenocarcinoma IC₅₀^MCF7 = 306.68 μg mL⁻¹ and cervix epitheloid carcinoma cell lines IC₅₀^HeLa = 315.49 μg mL⁻¹. Wild blackberry varieties had higher extraction yields, higher total polyphenolic contents and possessed stronger biological effects compared to cultivated blackberries (P < 0.05). All blackberry extracts showed high biological potential that could be attributed to high total polyphenols and flavonoids content and could be utilised as value-added functional food.     

Antioxidative and anti‐inflammatory potential of phenolics from purple basil (Ocimum basilicum L.) leaves induced by jasmonic,arachidonic and β‐aminobutyric acid elicitation

The study presents changes in the phenolic levels, antioxidant and anti‐inflammatory potential of purple basil leaves caused by different chemical elicitors: arachidonic acid (AA), jasmonic acid (JA) and β‐aminobutyric acid (BABA). The application of the all tested elicitors increased the concentration of phenolic compounds, including flavonoids and phenolic acids; especially, in comparison with control (457.62 μg g^?1 FW), the rosmarinic acid level significantly increased after AA and JA treatment ‐ 705.0 and 596.5 μg g^?1 FW, respectively. Phenolics from AA‐elicited plants showed the highest anti‐inflammatory activities designated as lipoxygenase (EC₅₀ = 1.67 mg FW mL^?1) and cyclooxygenase inhibition (EC₅₀ = 0.31 mg FW mL^?1). Elicitors' treatments (especially AA and JA) may be a very useful biochemical tool for improving the production of phenolic compounds in purple basil leaves.     

Efficacy of Acorus calamus L. essential oil as a safe plant‐based antioxidant,Aflatoxin B1 suppressor and broad spectrum antimicrobial against food‐infesting fungi

The study explores the efficacy of Acorus calamus L. essential oil (EO) as a safe plant‐based broad spectrum antifungal, antiaflatoxin, antioxidant food additive. The oil completely inhibited the growth and toxin production of the toxigenic strain of Aspergillus flavus at 0.4 and 0.25 μL mL^?1, respectively. EO exhibited pronounced antifungal activity against sixteen food‐infesting fungal species at 0.5 μL mL^?1. The EO showed strong antioxidant efficacy (IC₅₀ 1.06 μL mL^?1) and nonphytotoxic nature on germination of chickpea seeds. The EO was found nonmammalian toxic showing high LD₅₀ (4877.4 μL kg^?1) for mice (oral, acute). The chemical profile of EO was determined through GC and GC–MS analysis. The findings strengthen the possibility of A. calamus EO as a plant‐based food additive in view of its favourable safety profile, antioxidant and antiaflatoxigenic efficacy and broad spectrum antimicrobial activity against food‐infesting fungi.     

Solid‐phase extraction and spectrophotometric determination of Allura Red (E129) in foodstuff,soft drink,syrup and energy drink samples: a comparison study

Amberlite XAD‐7 and XAD‐8 resins were used as adsorbents for preconcentration and determination of Allura Red (AR) food dye in aqueous medium. The effects of pH, sample volume, sample and eluent flow rates on the extraction of AR were optimised. The determination of dye was performed at 506.0 nm using spectrophotometry. Interference effects of matrix ions and some dyes were also investigated under optimised conditions. The methods permitted low detection limits which were 1.2 and 0.6 μg L^?1 for XAD‐7 and XAD‐8, respectively. Adsorption behaviours were investigated by adsorption isotherms and zero charge pH experiments. Validations of the method were performed by determination of AR contents in some foodstuffs. AR contents of liquid samples were found between 58 and 440 μg mL^?1. AR concentrations of solid samples were between 416 and 432 μg g^?1. The XAD‐7 and XAD‐8 resins presented a fast and reliable potential to determine AR dye in real samples.     

Assessment of chemically characterised Rosmarinus officinalis L. essential oil and its major compounds as plant‐based preservative in food system based on their efficacy against food‐borne moulds and aflatoxin secretion and as antioxidant

The study explores antifungal, anti‐aflatoxigenic and antioxidant efficacy of Rosmarinus officinalis essential oil (ROEO) and its major compounds. In addition, the mode of action of ROEO and its practical efficacy as preservative have been assessed. GC‐MS analysis of ROEO identified 16 compounds; α‐pinene, 1,8‐cineole and camphor being the major compounds. The minimum concentration for inhibition of growth and aflatoxin B₁ secretion against A. flavus (LHP‐6) was found to be 1.5, >5.0, 4.0 and 3.0 μL mL^?1 and 1.25, >5.0, 3.5 and 3.0 μL mL^?1 for ROEO, α‐pinene, 1,8‐cineole and camphor, respectively. The IC₅₀ value through DPPH analysis and percentage inhibition of linoleic acid peroxidation of ROEO were 0.042 μL mL^?1 and 71.05%, respectively. The targeted site of antifungal action of ROEO was confirmed as plasma membrane through ergosterol measurement and TEM analysis. Moreover, ROEO significantly protected Piper nigrum fruits against mould infestation upto 6 months in in vivo trial.     

Bifurcaria bifurcata: a key macro‐alga as a source of bioactive compounds and functional ingredients

The aim of this work was to study the proximate composition and the bioactive profile of Bifurcaria bifurcata. It contains 73.31 ± 0.69% of moisture, 8.57 ± 0.11 g per 100 g dry weight (d.w.) of protein, 5.81 ± 0.14 g per 100 g d.w. of lipid content and 30.15 ± 0.00 g per 100 g d.w. of ash. The polyunsaturated fatty acids were the most abundant fatty acid (FA), accounting for 2426.56 mg per 100 g which represents 41.77% of the total FA. The methanolic fraction showed high quantity of polyphenols (220.01 ± 0.010 phloroglucinol equivalents g^?1 extract), DPPH radical reduction capacity (EC₅₀:58.82 μg mL^?1) and oxygen radical absorbent capacity (3151.35 ± 119.33 μmol Trolox equivalents g^?1 extract). The highest antimicrobial effect was observed against Pseudomonas aeruginosa (11.3 ± 1.5 mm) and Saccharomyces cerevisiae (IC₅₀:17.07 μg mL^?1) induced by methanolic and dichloromethane fractions, respectively. Dichloromethane fraction revealed the highest antitumor activity on Caco‐2 and HepG‐2 cells. Bifurcaria bifurcata can be a promising source of bioactive compounds and functional ingredients.