Chemical and biological variability of hot pepper fruits (Capsicum annuum var. acuminatum L.) in relation to maturity stage

The aim of the present work was to evaluate the chemical composition and the radical-scavenging and antioxidant activities of hot pepper fruits (Capsicum annuum L. var. acuminatum) at three maturity stages (small green, green and red). GC–MS analysis of n-hexane and chloroform fractions showed a different composition between the three stages of ripening. The first stage of maturation (small green) showed the highest radical-scavenging activity (IC₅₀ of 129 μg/ml). Using the bovine brain peroxidation assay, the methanolic extract of green pepper showed significant antioxidant activity (IC₅₀ of 522 μg/ml). Addition of methanolic extract of red and green pepper inhibited oxidation of linoleic acid. Methanolic extract of red pepper showed greater antioxidative potency than the others (IC₅₀ of 3 μg/ml). The different composition of lipophilic compounds and the various amount of phenolics, showed in the three stage of ripening of C. annuum var. acuminatum fruits, modifies the antioxidant activity.     

Angiotensin I-converting enzyme inhibitory activity of chickpea and pea protein hydrolysates

ACE inhibitory activity was studied for different hydrolysates obtained from protein concentrates of chickpea (kabuli and desi) and yellow pea (Golden) using in vitro gastrointestinal simulation, alcalase/flavourzyme, and papain. Protein/peptide profiles studied by SDS–PAGE and SE-HPLC, showed a rich composition of the hydrolysates in small peptides having MWs under 4 kDa. Papain hydrolysed yellow pea proteins showed the highest ACE inhibitory activity. In addition, chickpea desi proteins hydrolysed by in vitro gastrointestinal simulation showed higher ACE inhibition (IC₅₀ of 140 ± 1 μg/ml) compared to its digests obtained by alcalase/flavourzyme (IC₅₀ of 228 ± 3 μg/ml) or papain (IC₅₀ of 180 ± 1 μg/ml) and to chickpea kabuli hydrolysed by gastrointestinal simulation (IC₅₀ of 229 ± 1 μg/ml). The results demonstrate that enzymatic hydrolysates of chickpea and pea proteins contain bioactive ACE inhibitory peptides; furthermore, the type of enzyme used for hydrolysis affects the ACE inhibitory activity.     

Studies on the anthocyanin profile and biological properties from the fruits of Acanthopanax senticosus (Siberian Ginseng)

The anthocyanin profile and biological activities, including antioxidant, xanthine oxidase inhibitory, angiotensin converting enzyme (ACE) inhibitory, and anticancer of Acanthopanax senticosus fruits were evaluated for the first time. The acidified 80% methanol extract of this species exhibited high biological properties at a concentration of 60 μg/ml. Moreover, cyanidin-3-O-(2″-O-xylosyl)-glucoside was identified using C₁₈ column chromatography, NMR spectroscopy, and HPLC-DAD-ESI/MS analysis. This compound was present at 5.2 mg/g, representing approximately 91% of the total peak area and possessed strong antioxidant effects against DPPH, ABTS, and hydroxyl radicals (IC₅₀ of 85.2, 43.7, and 126.6 μg/ml, respectively). Cyanidin-3-O-(2″-O-xylosyl)-glucoside also exhibited significant inhibitory activities against xanthine oxidase and ACE (IC₅₀: 55.5 and 47.1 μg/ml, respectively). Especially, LNCap (prostate), MOLT-4F (leukaemia), and ACHN (renal) cell lines exhibited potent anticancer effects, with IC₅₀ of 5.2, 11.2, and 22.5 μg/ml, respectively, in comparison with other cancer cell lines. Therefore, A. senticosus fruit may be utilised as an effective source for food and nutraceutical uses due to its high anthocyanin content as well as various biological properties.     

Carotenoid content,its stability during drying and the antioxidant activity of commercial coriander (Coriandrum sativum L.) varieties

Although populations of several tropical countries are under severe vitamin-A deficiency, traditional sources of pro-vitamin-A carotenoids have not been exploited due to the lack of relevant research data. In this study, ten commercial varieties of coriander (Coriandrum sativum L.), grown at identical conditions, were evaluated for carotenoids, their bio-efficacy and stability during drying, with the main emphasis on β-carotene (vitamin-A precursor) analysis by HPLC-MS. In all varieties, β-carotene content was higher in foliage at mature stage, than in seedlings and seeds. Variety GS4 Multicut produced highest biomass (6.18 ± 0.73 g/plant), total carotenoids (217.50 ± 5.6 mg/100 g DW) and β-carotene (73.64 ± 0.3 mg/100 g DW) at pre-flowering stage. Carotenoids extract showed a high antioxidant activity with IC₅₀ value of 14.29 ± 1.68 μg/ml, scavenging hydroxyl radicals and rendering higher protection to DNA than by standard gallic acid (IC₅₀ = 357.21 ± 4.29). Microwave drying of foliage was rapid with better retention of pigments, high intactness of trans-β-carotene and higher extractability of pigments when compared with oven drying.     

Isolation and identification of an antiproliferative substance from fructose–tyrosine Maillard reaction products

We isolated a substance from fructose–tyrosine Maillard reaction products (MRPs) and investigated its antiproliferative effect on six human cancer cell lines. The ethyl acetate fraction of fructose–tyrosine MRPs showed a strong antiproliferative effect; this fraction was isolated and purified using silica gel column chromatography, semipreparative RP-HPLC, and recycling HPLC. The structure of the purified compound was determined using spectroscopic methods. The isolated compound was identified as 2,4-bis(p-hydroxyphenyl)-2-butenal (C₁₆H₁₄O₃, HPB242). HPB242 inhibited cell growth in a dose-dependent manner (10–80 μg/ml) on the six human cancer cell lines. The IC₅₀ values of HPB242 on the six human cancer cell lines were 17.34 μg/ml (MCF-7), 29.21 μg/ml (HCT-116), 34.57 μg/ml (H-460), 34.87 μg/ml (HepG2), 48.77 μg/ml (PC-3), and 55.83 μg/ml (MKN-45).     

Podocarpane,isopimarane, and abietane diterpenoids from Isodon lophanthoides var. graciliflorus

Four new diterpenoids including two podocarpanes, graciliflorins A (1) and B (2), an isopimarane acetal, graciliflorin C (3), and a rearranged abietane, graciliflorin D (4) were isolated from the aerial parts of Isodon lophanthoides var. graciliflorus (Lamiaceae) along with podocarpa-8,11,13-triene-3α,13-diol (5) and micranthin B (6). Their structures were elucidated based on the spectroscopic data. The in vitro cytotoxicity of compounds 1–4 and 6 against human carcinoma A549, MCF-7, and HeLa cell lines were evaluated using the MTT colourimetric assay. Micranthin B (6) showed moderate activity against all the cells with IC₅₀ values of 16.29, 18.20, and 22.25 μM, while compounds 1–4 were inactive (IC₅₀ > 50 μg/ml).     

Bioactive characteristics and antioxidant activities of nine peppers

The contents of bioactive compounds (Vitamin C (Vc), carotenoids, total phenolics, phenolic profiles and capsaicinoids) and antioxidant activities (reducing power, DPPH-scavenging activity and lipid peroxidation inhibition ability) of nine peppers from Yunnan province China, belonging to seven cultivars, were determined. Red and green Point pepper/Long-point pepper were from the same cultivars at different maturity stages. Results showed that all fresh pepper samples were rich in Vc, carotenoids, and total phenolics. Vc contents varied mainly depending on the pepper cultivars, except for the ripened stages. The phenolic content in red Fructus Capsici (Capsium frutescens L.) was significantly higher than that of others (Capsium annuum L.) (p < 0.05). Seven phenolics were identified in four pepper samples, while six phenolics were detected in other samples. The capsaicinoid contents of nine peppers showed a large variability, and the content in red Fructus Capsici was significantly higher than that of others (p < 0.05). The extracts of nine peppers showed high reducing power, DPPH-scavenging activity, and lipid peroxidation inhibition ability, and the activities of Fructus Capsici were significantly stronger than those of others (p < 0.05). Furthermore, antioxidant activities of nine peppers correlated well with their total phenolic contents.     

NMR-based quantification of rosmarinic and carnosic acids,GC–MS profile and bioactivity relevant to neurodegenerative disorders of Rosmarinus officinalis L. extracts

A comparative study of phytochemicals content and biological properties of eight Rosmarinus officinalis (rosemary) populations (RO1–RO8) collected in different areas of Tunisia was carried out. Two of the main rosemary constituents, rosmarinic and carnosic acids, were quantified by an NMR technique. Carnosic acid content was higher than that of rosmarinic acid. The non-polar constituents were examined by GC and GC–MS. Total phenols and flavonoids content were also determined in order to discuss the possible correlation between these phytochemicals and bioactivity. Antioxidant activity was investigated through different in vitro assays. Sample RO3 from a sub-humid area showed the highest potency in 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay (IC₅₀ of 6.74 μg/mL) while RO5 from arid area exhibited the highest radical 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) scavenging activity (IC₅₀ of 5.92 μg/mL). The most active sample in Ferric Reducing Antioxidant Power (FRAP) assay was RO4 (62.21 μM Fe(II)/g), followed by RO1–RO3 and RO8 harvested in sub-humid areas with values in the range 49.61–58.72 μM Fe(II)/g. Extracts were able to inhibit acethylcholinesterase (AChE) and butyrylcholinesterase (BChE). Sample RO5 exhibited a promising AChE inhibitory activity (IC₅₀ of 4.47 μg/mL). Thus, we suggest rosemary extracts as a functional food ingredient for preventing AD.     

Purification and characterization of an antioxidant protein from Ginkgo biloba seeds

A new antioxidation protein named G4b was purified from Ginkgo biloba seed albumin. The results of chemical colorimetry and chemiluminescence methods showed that the hemi-inhibitory concentration (IC₅₀) of G4b on the hydroxyl radical were 78.1 μg/mL and 115.1 μg/mL, and the IC₅₀ on DPPH was 100.7 μg/mL, while the IC₅₀ on the superoxide anion and DNA injury were 23 μg/mL and 27 μg/mL, respectively, and the abundance of sulfur amino acids and aromatic amino acids contained in G4b might contribute to its strong antioxidation properties. G4b is a kind of homogeneous and novel protein with a molecular mass of 29,247 Da, consisting of two peptides with similar molecular weights linked by a disulfide bond. Result of β-elimination reaction showed that G4b is connected with a small quantity of polysaccharide by an oxygen-glucosidic bond.     

Purification and characterization of angiotensin I-converting enzyme inhibitory peptides of small red bean (Phaseolus vulgaris) hydrolysates

Angiotensin I-converting enzyme (ACE) inhibitory activity was investigated for small red bean (Phaseolus vulgaris) protein hydrolysate produced by sequential digestion of Alcalase, papain followed by in vitro gastrointestinal simulation. The hydrolysate had ACE inhibitory activity with IC₅₀ of 67.2 ± 1.8 μg protein/mL. Peptides responsible for potent ACE inhibitory activity were isolated by a three-step purification process, including ultrafiltration, gel filtration and preparative reverse phase high performance chromatography (RP-HPLC). The fraction obtained after RP-HPLC fractionation with the highest activity yielded an IC₅₀ of 19.3 ± 1.4 μg protein/mL. Enzymatic kinetic studies using this fraction demonstrated competitive inhibition with K_i of 11.6 ± 1.7 μg protein/mL. Mass spectrometric characterization identified for the first time the octapeptide PVNNPQIH which demonstrated an IC₅₀ value of 206.7 ± 3.9 μM. The results expand the knowledge base of ACE inhibitory properties of small red bean protein hydrolysate and should be useful in further identification of specific ACE inhibitory peptides in beans.     

Cyanidin-3-glucoside,nutritionally important constituents and in vitro antioxidant activities of Santalum album L. berries

Anthocyanins are ubiquitous plant pigments. They are hydrophilic, and have high tinctorial value hence, used as natural food colorant. The main aim of this study was characterization of Santalum album L. berries for its pigment content, nutritionally important phytoconstituents and functional attributes. Major pigment identified was cyanidin-3-glucoside (0.21% FW) as confirmed by spectral characteristics including LC–MS. Nutrients such as total carbohydrates (8.5), proteins (5.8), free amino acids (0.8 g), oil (1.5 g), ascorbic acid (1.5 μg), tocopherols (0.3 mg) and niacin (5.2 mg) per 100 g berries were determined along with total soluble solids (13.4°Brix) and phenols (3.1 mg GAE/g). Reducing power and DPPH· scavenging assays showed highest activity in methanol extract, which also efficiently protected bleaching of β-carotene (EC₅₀ 285 μg/mL) in β-carotene–linoleate model. Pigment rich crude extract was not toxic to HepG2 cells during 24 h exposure, whereas cyanidin-3-glucoside was cytotoxic (IC₅₀ 0.1 μg/mL).     

The influence of fruit ripening on the phytochemical content and biological activity of Capsicum chinense Jacq. cv Habanero

During the past decade, it has been reported that the consumption of certain foods and spices such as pepper may have a positive effect on health. The present study evaluates the influence of fruit ripening on total phenols, flavonoids, carotenoids and capsaicinoids content and antioxidant, hypoglycaemic and anticholinesterase activities of Capsicum chinense Jacq. cv Habanero. The chemical investigation showed a different composition between the two stages of ripening (immature and mature). Generally, the concentration of carotenoids and capsaicinoids increased as the peppers reached maturity, whereas the concentration of phenols declined. The immature fruits showed the highest radical scavenging activity (IC₅₀ of 97.14 μg/ml). On the contrary, the antioxidant activity evaluated by the β-carotene bleaching test showed a significant activity for mature peppers (IC₅₀ value of 4.57 μg/ml after 30 min of incubation). Mature peppers inhibited α-amylase with an IC₅₀ of 130.67 μg/ml. The lipophilic fractions of both mature and immature peppers exhibited an interesting and selective inhibitory activity against α-amylase with IC₅₀ values of 29.58 and 9.88 μg/ml, respectively. Both total extracts of mature and immature peppers inhibited butyrylcholinesterase selectively. The obtained results underline the potential health benefits as a result of consuming C. chinense Habanero and suggest that it could be used as new valuable flavour with functional properties for food or nutriceutical products on the basis of the high content of phytochemicals and found biological properties.     

Identification and characterisation of coumarins from the roots of Angelica dahurica and their inhibitory effects against cholinesterase

The current research was the first to investigate coumarins and their cholinesterase inhibitory activities from Angelica dahurica roots. The ethanol extract of this species (100 μg/ml) possessed inhibitory effects against human acetylcholinesterase (hAChE) and butylcholinesterase (BChE) with IC₅₀ values of 26.40 and 14.71 μg/ml, respectively. To evaluate the compounds responsible for these activities, the ethanol extract was chromatographed, which yielded ten coumarins, including isoimperatorin (1), imperatorin (2), senbyakangelicol (3), oxypeucedanin (4), byakangelicol (5), t-OMe-oxypeucedanin hydrate (6), t-OMe-byakangelicin (7), angelol H (8), byakangelicin (9), and oxypeucedanin hydrate (10). Among them, coumarin 5, 7, and 9 showed potent inhibition with IC₅₀s of 46.3, 39.0, and 55.4 μM (hAChE) as well as 45.2, 25.6, and 42.4 μM (BChE), respectively. Moreover, their inhibition modes against two cholinesterases exhibited noncompetitive. The individual coumarin contents were remarkable differences, especially, oxypeucedanin (4) was the most predominant compound (8710.9 μg/g), representing approximately 55.0% of the total content.     

Pomegranate husk extract,punicalagin and ellagic acid inhibit fatty acid synthase and adipogenesis of 3T3-L1 adipocyte

Fatty acid synthase (FAS) has been recognized as a potential therapeutic target for obesity. In this study, for the first time, the inhibitory effect of pomegranate husk extract, punicalagin and ellagic acid on FAS was investigated. We found them potently inhibiting the activity of FAS with half-inhibitory concentration values (IC₅₀) of 4.1 μg/ml (pomegranate husk extract), 4.2 μg/ml (4.50 μM, punicalagin) and 1.31 μg/ml (4.34 μM, ellagic acid), respectively. Moreover, they all exhibited time-dependent inactivation of FAS. Punicalagin and ellagic acid inhibited FAS with different mechanisms compared to previously reported inhibitors, through inactivating acetyl/malonyl transferase and β-ketoacyl synthase domains, respectively. Additionally, 100 μg/ml pomegranate husk extract, 5.24 μg/ml (5 μM) punicalagin and 4.5 μg/ml (15 μM) ellagic acid effectively reduced lipid accumulation inside FAS over-expressed 3T3-L1 adipocytes. Since FAS plays a key role in the biosynthesis pathway of fatty acid, these findings suggest that pomegranate husk extract, punicalagin and ellagic acid have potential in the prevention and treatment of obesity.     

Cancer preventive effect of Thai rat-tailed radish (Raphanus sativus L. var. caudatus Alef)

The cancer preventive effect of Raphanus sativus L. var. caudatus Alef (Thai rat-tailed radish) extract was evaluated against the HCT116 colon cancer cell line. HPLC and GC–MS were used to identify the phytoconstituents in Thai rat-tailed radish extract. Thai rat-tailed radish extract showed high cytotoxicity against HCT116 with an IC₅₀ at 9.42 ± 0.46 μg/ml. Apoptosis induced by Thai rat-tailed radish extract was confirmed (a) by DAPI staining which demonstrated nuclei morphological changes, chromatin condensation and nuclear fragmentation and (b) by Annexin V/PI flow cytometry. GC–MS analysis of Thai rat-tailed radish extract revealed two isothiocyanates; sulforaphane and sulforaphene, whose cytotoxicity and apoptosis induction effect were evident, giving Thai rat-tailed radish extract its anticancer attributes. This is the first report on the cancer preventive effect of Thai rat-tailed radish extract against HCT116 colon cancer cell line, through apoptosis induction.     

Yerba mate tea (Ilex paraguariensis): Phenolics,antioxidant capacity and in vitro inhibition of colon cancer cell proliferation

Mate tea (MT) is a rich source of phenolic compounds that vary depending on geographical origin and mode of preparation. Total phenolic concentration and antioxidant capacity of Mate tea (Ilex paraguariensis) products were determined in this work. In addition, a representative MT was tested for in vitro inhibition of human colon carcinoma cell proliferation. Total polyphenol concentration, was measured using Folin–Ciocalteau method, ranged from 90 to 176 mg gallic acid eq (GAE)/g dry leaves (DL) in traditional MT and from 40 to 113 mg GAE/g DL in MT added with other flavouring ingredients. It was estimated that a cup of tea (250 ml) containing one teaspoon (5 g) of instant MT could provide an average intake of 1.5 g GAE. Fresh tea (FT) from Mate leaves displayed high antioxidant capacity (85 ± 1%) and preferentially inhibited 50% of net growth of human colorectal adenocarcinoma cells CaCo-2 (GI₅₀ = 1.0 ± 0.03 μg/ml) and HT-29 (GI₅₀ = 105.2 ± 15.2 μg/ml) when compared with the CCD-33Co normal colon fibroblast cell line (GI₅₀ > 300 μg/ml). MT inhibited in vitro colon cancer cell proliferation possibly mediated via pro-oxidant activities, therefore represents a potential source of chemopreventive agents that deserve further investigation.     

Identification of bioactive peptides after digestion of human milk and infant formula with pepsin and pancreatin

Seven human milks were subjected to an in vitro digestion with pepsin and pancreatin to identify the peptides released from human proteins. On the basis of their sequences, 11 of the 23 peptides were synthesised and their angiotensin converting enzyme (ACE)-inhibitory and antioxidant activities were measured. The β-casein peptides HLPLP and WSVPQPK showed potent ACE-inhibitory and antioxidant activity, with a protein concentration needed to inhibit 50% ACE activity (IC₅₀) of 21 μm and a Trolox Equivalent Antioxidant Capacity (TEAC) of 1.297 μmol 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) equivs μmol⁻¹ of peptide, respectively. These activities were determined after digestion of eight infant formulas and compared with those found in digested human milk. One of the infant formulas exhibited a low IC₅₀ value (60.11 μg protein mL⁻¹ of reconstituted formula) and a high TEAC value (1.7056 μmol Trolox equivs mg⁻¹ of protein) and was therefore selected to identify the peptides responsible of these activities.     

Glycine max (L.) Merr., Vigna radiata L. and Medicago sativa L. sprouts: A natural source of bioactive compounds

The consumption of sprouts, common in Asia, has been growing in western countries, once they are a natural healthy food and considered as a valuable dietary supplement. Comparing with their mature counterparts, sprouts are usually richer in health-promoting phytochemicals. So, the nutritional composition and the biological potential of widely consumed sprouts of three species – Glycine max (L.) Merr., Vigna radiata L. and Medicago sativa L. – were compared for the first time. Phenolic compounds and phytosterols were analyzed by HPLC–DAD and organic acids by HPLC–UV. The volatile profile was determined by HS-SPME/GC–IT/MS. Fourteen phenolic compounds (including four isoflavones), three sterols one triterpene, sixteen fatty acids, seven organic acids and thirty volatile compounds were determined. The antioxidant activity was assessed against DPPH^?, superoxide and nitric oxide radicals. G. max sprouts were the most active against DPPH^? (IC₅₀ = 1.337 mg/mL), while those of M. sativa were the most effective against superoxide and nitric oxide radicals (IC₅₀ = 67 μg/mL and IC₅₀ = 426 μg/mL, respectively). Data provide evidence of great similarities between G. max and M. sativa sprouts, both being rich in phenolic compounds, fatty acids and volatiles, and exhibiting better antioxidant activity. On the other hand, V. radiata showed higher amounts of sterols, triterpenes and organic acids. In this study it was found that the sprouts are a good source of bioactive compounds in our diet with health-promoting antioxidants.     

Licochalcone A: A lipase inhibitor from the roots of Glycyrrhiza uralensis

In order to develop a functional food and anti-obesity drug through the inhibition of absorption of dietary lipids, we investigated the inhibitory effects on pancreatic lipase of extracts from more than 800 species of herbs in Korea. In this study, licochalcone A was isolated from the ethyl acetate/n-hexane fraction of ethyl acetate extract of the roots of Glycyrrhiza uralensis. Its structure was elucidated by ¹H NMR, ¹³C NMR and HR-EI mass spectroscopy. Licochalcone A substantially inhibited activity with IC₅₀ value of 35 μg/mL (103.4 μM). In an investigation of reversibility, licochalcone A was shown to be reversible to pancreatic lipase. The inhibition mode of licochalcone A was a non-competitive inhibitor, determined by Lineweaver–Burk plot analysis, and K_i value was 11.2 μg/mL (32.8 μM). Furthermore, licochalcone A significantly reduced the production of oleic acid with not only artificial substrate DNPB but natural substrate triolein by pancreatic lipase.     

Cytochrome P450 inhibitory potential of selected Indian spices — possible food drug interaction

Spices constitute an important group of food which is virtually indispensable in the culinary art. In a view, these spices feared to pose a probability to affect the disposition of conventional pharmaceuticals through inhibition of human cytochrome P450 (CYPs) enzymes. In the present study an approach has been made to evaluate the possible CYP inhibition potential with some Indian spices (Capsicum annuum, Murraya koenigii, Zingiber officinale) and their major bioactive compounds, in combination with pooled microsome; as well as commercially available recombinant human CYP3A4, CYP2D6, CYP2C9 and CYP1A2. Quantification of the bioactive compound was determined through RP-HPLC, in order to standardize the plant material. CYP–carbon monoxide (CYP–CO) complex assay result indicated that all the plants and their bioactive compounds have an interaction potential with CYPs. Fluoregenic assay results indicated that the spice extracts have higher inhibition potential comparing to their single bioactive molecule. The higher enzyme inhibition potential by the extracts may be related to the synergistic effects due to the presence of other constituents in the extract. Capsaicin and C. annuum showed the lowest IC₅₀ value and 6-gingerol and Z. officinale extract showed the highest IC₅₀ value among the entire sample tested. The entire sample showed significantly less (P < 0.001, P < 0.01) interaction potential than known inhibitors. These findings indicate that selected spices are unlikely to cause clinically relevant drug interactions involving the inhibition of major CYP isozymes.