Antitumour properties of the leaf essential oil of Xylopia frutescens Aubl. (Annonaceae)

The aim of this study was to investigate the chemical composition and anticancer effect of the leaf essential oil of Xylopia frutescens in experimental models. The chemical composition of the essential oil was analysed by GC/FID and GC/MS. In vitro cytotoxic activity of the essential oil was determined on cultured tumour cells. In vivo antitumour activity was assessed in Sarcoma 180-bearing mice. The major compounds identified were (E)-caryophyllene (31.48%), bicyclogermacrene (15.13%), germacrene D (9.66%), δ-cadinene (5.44%), viridiflorene (5.09%) and α-copaene (4.35%). In vitro study of the essential oil displayed cytotoxicity on tumour cell lines and showed IC₅₀ values ranging from 24.6 to 40.0 μg/ml for the NCI-H358M and PC-3M cell lines, respectively. In the in vivo antitumour study, tumour growth inhibition rates were 31.0–37.5%. In summary, the essential oil was dominated by sesquiterpene constituents and has some interesting anticancer activity.     

Bioassay-guided isolation and identification of antifungal components from propolis against Penicillium italicum

The present study was aimed at identification of antifungal components against Penicillium italicum from Chinese propolis with bioassay-guided fractionation technique. Propolis ethanolic extract (PEE) was separated and purified by liquid–liquid extraction and thin layer chromatography (TLC) and the most active band was subjected to HPLC–MS/MS to identify the antifungal compounds. The results showed PEE and its fractions had strong antifungal activity against P. italicum. Among the fractions of PEE partitioned by petroleum ether, ethyl acetate, n-butanol and water, ethyl acetate fraction (E-Fr) exhibited the most effective activity against P. italicum. Further bioautographic TLC assay showed Band I, with Rf value of 0.70, had an inhibitive zone, which showed the strongest antifungal activity and completely inhibited the growth of P.italicum at 200 mg/L. Bioactive components found in Band I were further identified as pinobanksin, pinocembrine, chrysin and galangin. This study exhibited Chinese propolis and its main flavonoids was potential natural alternatives for the control of citrus blue mould caused by P.italicum.     

Identification and quantitative determination of glucosinolates in seeds and edible parts of Korean Chinese cabbage

Glucosinolates (GSLs) have attracted major interest due to the chemopreventive properties of some of their transformation products. GSLs in the seeds and edible parts of Korean Chinese cabbage (Brassicacampestris L. ssp. pekinensis) were identified and quantified by LC–ESI–MS and LC–UV. As a result, nine GSLs were identified: progoitrin, glucoraphanin, glucoalyssin, gluconapin, 4-hydroxy-3-indolylmethyl, glucobrassicanapin, glucoerucin, glucobrassicin, and 4-methoxyglucobrassicin. The total GSL levels were 268–198 and 23.0–15.8 μmol/g dry weight (DW) for seeds and edible parts, respectively. Gluconapin (197 μmol/g DW) was the highest individual GSL in seeds, whereas 4-methoxyglucobrassicin (6.08–4.94 μmol/g DW) and glucobrassicanapin (8.18–3.09 μmol/g DW) were found in the edible parts. In addition, LC–MS profiles of the nine GSLs identified from Korean Chinese cabbage were subjected to principal components analysis (PCA) to evaluate differences among samples. The metabolome among the four cultivar seed or edible parts was clearly separated by PCA.     

Characterisation of volatiles and polyphenols for quality assessment of alcoholic beverages prepared from Corsican Myrtus communis berries

In the present work, the essential oil compositions of Myrtus communis berries from ten Corsican localities were studied and no chemical variability was observed. HS–SPME, GC and GC/MS analysis were carried out for the characterisation of volatile fractions of M. communis berries and two derived commercial alcoholic beverages (liqueur and eau-de-vie). Quantitative variations due to the distillation process were observed between the two beverages. The volatile compositions of Corsican myrtle alcoholic products were characterised by high amounts of monoterpene hydrocarbons and oxygenated monoterpenes with α-pinene and 1,8-cineole as major components. The polyphenolic compositions of the berry extract and the liqueur were also established using HPLC–DAD and LC–MS–MS; high concentrations of flavonol glycosides, flavonols and flavanols were reported.     

Rapid screening and identification of antioxidants in aqueous extracts of Houttuynia cordata using LC–ESI–MS coupled with DPPH assay

Houttuynia cordata Thunb. has been used traditionally as immune stimulant and anticancer agent. An aqueous extract of H. cordata tea showed high radical scavenging activity determined by off-line DPPH assays. Then, it was screened for its antioxidant components via an on-line DPPH radical scavenging technique coupled with a liquid chromatography–electrospray ionization mass spectrometer (LC–ESI–MS). Based on their mass spectra and fragmentation patterns; the antioxidant compounds were identified as quinic acid derivative, caffeic acid derivatives, procyanidin B, neo-chlorogenic acid, catechin, chlorogenic acid, crypto-chlorogenic acid and quercetin hexoside. LC–MS/MS in multiple reactions monitoring (MRM) mode was used to quantify these antioxidant compounds. Chlorogenic acid was found to be a major component in H. cordata tea.     

Characterisation of phenolic antioxidants in aqueous extract of Orthosiphon grandiflorus tea by LC–ESI-MS/MS coupled to DPPH assay

Aqueous extract from Orthosiphon grandiflorus tea was on-line screened for its antioxidant components based on their capacity to scavenge free DPPH (1,1-diphenyl-2-picrylhydrazyl) radical after separation by a LC gradient. The structural elucidation of the active compounds was achieved by negative ionisation LC–ESI-MS/MS. Based on their mass spectra and fragmentation patterns related to antioxidant activity trace, seven compounds showing strong DPPH scavenging were identified to be danshensu, caffeic acid, caftaric acid, rosmarinic acid, caffeic acid derivative, sagerinic acid and salvianolic acid B. It is noted that danshensu, caftaric acid, sagerinic acid and salvianolic acid B have been reported in O. grandiflorus for the first time. In addition, the quantification of antioxidant compounds was performed using LC–MS/MS in multiple reaction monitoring (MRM) mode. Rosmarinic acid was found as a major component responsible for the antioxidant activity of this plant extract.     

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

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

Rapid quantitation of curcumin in turmeric via NMR and LC–tandem mass spectrometry

A rapid, sensitive and accurate ¹H NMR method has been developed for the quantitation of curcumin isolated from Curcuma longa rhizome (turmeric) extract, the results of which were compared with a validated LC–MS/MS method. The relative standard deviations of the methods were found to be 2.49% and 3.48% for the ¹H NMR and LC–MS/MS methods, respectively. The correlation coefficients were 0.998 for ¹H NMR and 0.995 for LC–MS/MS assay in the calibration range. The recoveries at 5 mg mL⁻¹ and 50 μg mL⁻¹ concentrations averaged to 99–101% for both techniques, respectively. The uncertainty of the measurement of curcumin via ¹H NMR spectroscopy was determined to be 5.80% while in LC–ESI-MS/MS method was 7.38%.     

Effect of bengkoang (Pachyrhizus erosus) fiber extract on murine macrophage-like J774.1 cells and mouse peritoneal macrophages

Bengkoang (Pachyrhizus erosus (L) Urban) is an edible root vegetable containing fairly large amounts of carbohydrates and crude fibers. The purpose of this study is to evaluate the effects of the bengkoang fiber extract (BFE) on macrophages in vitro and in vivo. BEF was prepared by heat-extraction from the bengkoang fiber in distilled water at 121 °C for 20 min. BFE stimulated the phagocytotic activity of J774.1 cells. In addition, BFE significantly facilitated production of tumor necrosis factor (TNF)-α and interleukin (IL)-6 by J774.1 cells and mouse peritoneal macrophages. BFE also facilitated the gene expression levels of inducible nitric oxide synthase and cyclooxygenase-2. The phagocytotic activity of mouse peritoneal macrophages was significantly enhanced by oral administration of BFE to BALB/c mice for 7 days. Ex vivo analysis revealed that production and gene expression of TNF-α and IL-6 in peritoneal macrophages from the BFE-administered mice were significantly enhanced. These results demonstrated that BFE activates macrophages in vitro and in vivo.     

The impact of fermentation and in vitro digestion on formation angiotensin converting enzyme (ACE) inhibitory peptides from pea proteins

Pea seeds were fermented by Lactobacillus plantarum 299v in monoculture under different time and temperature conditions and the fermented products were digested in vitro under gastrointestinal conditions. After fermentation and digestion ACE inhibitory activity was determined. In all samples after fermentation no ACE inhibitory activity was noted. Potentially antihypertensive peptides were released during in vitro digestion. The highest DH (68.62%) were noted for control sample, although the lowest IC₅₀ value (0.19 mg/ml) was determined for product after 7 days fermentation at 22 °C. The hydrolysate characterised by the highest ACE inhibitory activity was separated on Sephadex G10 and two peptides fractions were obtained. The highest ACE inhibitory activity (IC₅₀ = 64.04 μg/ml) for the first fraction was noted. This fraction was separated by HPLC and identified by LC–MS/MS and the sequence of peptide derived from pea proteins was determined as KEDDEEEEQGEEE.     

Hepatoprotective and antioxidant activity of ethanolic extracts of edible lotus (Nelumbo nucifera Gaertn.) leaves

Lotus (Nelumbo nucifera Gaertn.), an aquatic vegetable, is extensively cultivated in eastern Asia, particularly in China. An ethanolic extract of the leaves was studied for its hepatoprotective activity against CCl₄-induced liver toxicity in rats. In vitro and in vivo antioxidant activity was also assessed. The results showed the hepatoprotective activity of lotus leaf extract (LLE) at doses of 300 and 500 mg/kg and in vivo antioxidant activity at 100 mg/kg that was comparable with that of a standard treatment comprising 100 mg/kg of silymarin, a known hepatoprotective drug. These data were supplemented with histopathological studies of rat liver sections. The main flavonoids and phenolic compounds of LLE were analysed by HPLC-DAD-ESI/MS methods. Six of the compounds detected were tentatively characterised, one as catechin glycoside and five as flavonoid glycoside derivatives: miricitrin-3-O-glucoside, hyperin, isoquercitrin, quercetin-3-O-rhamnoside and astragalin.     

Assessment of the antioxidant potential of selected plant extracts – In vitro and in vivo experiments on pork

The antioxidant potential of selected plant extracts was assessed in vitro and in vivo experiments on pork. In the in vitro experiment, the anti-oxidative capacity of ethanol–water extract of Melissa officinalis (MW), ethanol–propylene–glycol extracts of M. officinalis (MP), Origanum vulgaris (O) and Salvia officinalis (S) at different dilutions was analysed. Furthermore a 2% essential oil concentrate was added to Origanum (OSi) and Salvia (SSi). In the two in vivo experiments in total 104 Slovak White Meaty pigs were fed with plant extracts (MW and O) at different doses with/without additional vitamin E.     

Antimicrobial potential of Coriandrum sativum L. against different Candida species in vitro

The aim of this study was to evaluate the chemical and antifungal activity of the essential oil from Coriandrum sativum L. (Apiaceae) against different Candida species. The essential oil (EO) was obtained by hydrodistillation and submitted to dry-column chromatography, resulting in six fractions, which were then submitted to TLC and GC–MS analysis. The main compounds identified were alcohols: 1-decanol (24.20%); 2E-decenol (18.00%); 2Z-dodecenol (17.60%); and aldehydes (89%). Antibacterial activity of the EO and its fractions was tested against five species of Candida albicans. The EO showed antimicrobial activity against all the species of Candida tested, except for Candida tropicalis CBS 94. Fractions 4 and 6 had a greater antibiotic spectrum, probably due to the presence of such alcohols as 3-hexenol, 1-decanol, 2E-decenol and 2Z-dodecenol. In conclusion, the EO and its fractions could be used as potential antimicrobial agents to treat or prevent Candida yeast infections.     

The specific chemical profile of Mediterranean propolis from Malta

Seventeen Maltese propolis samples were studied by GC–MS after silylation. They exhibited the typical Mediterranean chemical profile, rich in diterpene compounds (18–92% of TIC, GC–MS): 32 individual diterpenes were identified; 22 of them were present in each specimen. The other abundant compound group was that of sugars and sugar derivatives. In some samples, however, another compound group was observed (0–12% of TIC, GC–MS); the corresponding mass spectra were consistent with mono- and sesquiterpenyl esters of substituted benzoic acids. Two new propolis constituents of this group, daucane diterpene esters of hydroxybenzoic acids, were isolated. Their origin is suggested to be Ferula communis, as they are taxonomic markers for this species. All propolis samples were active against Staphylococcus aureus but only those with high concentrations of terpenyl esters showed antifungal activity against Candida albicans. The present results confirm that Mediterranean propolis is a valuable natural product with potential to improve human health.     

Contribution of Leu and Hyp residues to antioxidant and ACE-inhibitory activities of peptide sequences isolated from squid gelatin hydrolysate

Squid gelatin obtained from inner and outer tunics was hydrolysed with Alcalase to isolate antioxidant peptide sequences. The ACE-inhibitory activity of the isolated peptides was also evaluated. After fractionation by ultrafiltration and size-exclusion chromatography into four fractions, the antioxidant activity of the peptide fractions was determined by radical scavenging ability and ferric reducing power. Fraction FIII showed the highest antioxidant activity, although slight differences could be expected in the antioxidant activity of the different fractions based on the amino acid composition. FIII was subjected to liquid chromatography and tandem mass spectrometry (LC–MS/MS) and two major compounds were identified: the compound with m/z 952.42, which could be mostly comprised by the carbohydrate fucose, and the peptide with m/z 1410.63. Three possible sequences were proposed and synthesised for this peptide, and the contribution of Leu or Hyp residues to the antioxidant and ACE-inhibitory activities of the resulting sequence was evaluated. The presence of Leu residues in the peptide sequence in replacement of Hyp seems to play an important role in the antioxidant and ACE-inhibitory activity.     

Evolution of tebuconazole residues through the winemaking process of Mencía grapes

The removal of tebuconazole residues during the winemaking process, applied initially to red must elaborated from Mencía variety grapes from A.O.C. Valdeorrras (Ourense, N.W. Spain), was studied. Analytical determination of tebuconazole residues in grapes, musts and wines were perfomed by gas chromatography equipped with an ion trap mass spectrometry detector (GC–ITMS). Tebuconazole is retained on the solid matter (cakes and lees) and clarification agent. The eliminated percentage of tebuconazole in the final wine is 86%. The influence of this fungicide on the fermentative activity of Saccharomyces cerevisiae yeast and the Oenococcus oeni was also studied through in vitro assays. Liquid chromatography equipped with triple quadrupole mass spectrometer (LC–MS/MS) was used to determine tebuconazole residues in synthetic must and wine used for in vitro assays. No effect on the alcoholic or malolactic fermentation was observed; and no degradation of tebuconazole originated by these microorganisms was registered.     

Quali-quantitative analysis of flavonoids of Cornus mas L. (Cornaceae) fruits

The methanol extract obtained from the ripe fruits of Cornus mas L. (Cornaceae) have been phytochemically studied. On the basis of HPLC–PDA–MS/MSⁿ analysis eight compounds have been identified as quercetin, kaempferol, and aromadendrin glycosilated derivatives. Three major compounds have been also isolated by Sephadex LH-20 column chromatography followed by HPLC and characterised by NMR spectroscopy. Moreover, LC–PDA–MS analysis of the red pigment revealed the presence of three anthocyanins. The quantitative analysis of all compounds was reported.     

Chemopreventive properties of the bran extracted from a newly-developed Thai rice: The Riceberry

The potential anti-cancer activity of compounds extracted from Riceberry bran was evaluated in human cancer cell lines (Caco-2, MCF-7 and HL-60). Anti-proliferation and BrdU incorporation assays indicated a time–dose dependent effect of dichloromethane (DCM) and methanol (MeOH) extracts, and that HL-60 was the most sensitive cell. DNA fragmentation assay revealed that both extracts could induce different degrees of apoptosis. The apoptotic induction pathway of each extract determined by flow cytometry and immunoblotting assays revealed various phases of cell cycle arrest with alteration of pro-apoptotic p53, caspase-3, and cyclin proteins. The bioactive compounds in each extract were chemically analysed by GC–MS and LC–ESI–MS/MS. Results revealed the presence of two major anthocyanins, cyanidin-3-glucoside and peonidin-3-glucoside, in the MeOH extract, while the DCM extract contained higher content of plant sterols. The latter constituents are considered the major contributors to apoptotic mechanism in the sensitive cell. These bran products are worth developing into medicinal supplements.     

HPLC–MS analyses and bioactivities of novel chemicals in Devil’s club (Oplopanax horridus (Sm.) Miq.)

Devil’s club (Oplopanax horridus (Sm.) Miq.) has traditionally been used as a folk medicine by native North Americans for the treatment of various chronic diseases. A methanolic extract and its sub-fractions by liquid–liquid re-extraction, i.e., chloroform (CHCl₃), ethyl acetate (EtOAc), n-butanol (BuOH), and water fractions, were analysed by their anti-cancer activity with in vitro cell proliferative bioassays, and their antioxidant capacities in terms of the determination of total phenolic content (TPC), ORAC value, and DPPH free radical scavenging activity. The whole extract of the dried root contained high TPC and possessed strong ORAC and DPPH radical scavenging activities. In addition, the extract exhibited a strong anti-proliferative ability against HT-29 cancer cells, e.g., with an inhibitive rate of 56.5% with a 0.2-mg/mL extract. Further investigation by HPLC–UV–MS identified significant bioactive phenolic compounds in the chloroform fraction, including gallic acid, caffeic acid, 4-O-feruloylquinic acid, 5-O-feruloylquinic acid, ferulic acid, methyl feruloylquinate, methyl ferulate, and quercetin. These results suggested that the associated bioactivity of the plant might result from the contribution of phenolic compounds.     

Antioxidant activity and phenolic content of phenolic rich fractions obtained from black cumin (Nigella sativa) seedcake

The antioxidant activities of crude methanolic extract (CME) and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black cumin seedcake were investigated. DPPH radical scavenging activity, β-carotene–linoleate bleaching, and inhibition of corn oil oxidation were used to evaluate the antioxidant capacity. The total phenolics were found to be 78.8, 27.8, 32.1 and 12.1 mg gallic acid equivalents (GAE)/g in EAF, CME, WF and HF, respectively. The CME and EAF exhibited the highest DPPH followed by WF and HF. The extract/fractions showed high effect on reducing the oxidation of β-carotene. The effect of extract/fractions on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The oil peroxide and anisidine values were generally lower with addition of PRFs in comparison to a control. The predominant phenolic compounds identified by HPLC–DAD in CME and WF of black cumin seedcake were hydroxybenzoic, syringic and p-cumaric acids.