Phenolic profiles of cultivated, in vitro cultured and commercial samples of Melissa officinalis L. infusions

Melissa officinalis L. (lemon balm) is normally consumed as an infusion and presents therapeutic properties, such as sedative, carminative and antispasmodic, also being included in some pharmaceutical preparations. The phenolic profiles of different samples of lemon balm, prepared as infusions, were evaluated by HPLC-DAD-ESI/MS. The profiles were compared in order to understand the differences between cultivated, in vitro cultured and commercial (bags and granulated) samples. All the samples showed a similar phenolic profile, presenting differences only in the quantities found of each compound. Rosmarinic acid was the most abundant compound, being higher in commercial samples, especially in tea bag sample (55.68 mg/g of infusion) and lower in in vitro cultured sample (15.46 mg/g). Moreover, dimers, trimers and tetramers of caffeic acid were identified and quantified for the first time in lemon balm. Only one flavonoid, luteolin-3′-O-glucuronide was found in all the samples, ranging from 8.43 mg/g in commercial granulate sample to 1.22 mg/g in in vitro cultured sample. Overall, cultivated and in vitro cultured samples presented the lowest amounts of phenolic compounds (59.59 and 30.21 mg/g, respectively); otherwise, commercial samples showed the highest contents (109.24 mg/g for tea bag and 101.03 mg/g for granulate sample). The present study shows that infusion of lemon balm can be a source of phenolic compounds, known for their bioactive effects.     

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 in vitro and in vivo antioxidant properties of seabuckthorn (Hippophae rhamnoides L.) seed oil

The antioxidant capacity of seabuckthorn (Hippophae rhamnoides L.) seed oil was investigated with a number of established in vitro assays and in an in vivo study of carbon tetrachloride (CCl₄)-induced oxidative stress in mice. The results showed that DPPH radical scavenging activity, ferrous ion chelating activity, reducing power and inhibition of lipid peroxidation activity all increased with increasing concentrations of seabuckthorn seed oil. Moreover, the EC₅₀ values of seabuckthorn seed oil from the hydrogen peroxide, superoxide radical, hydroxyl radical scavenging assays were 2.63, 2.16 and 0.77 mg/ml, respectively. In the in vivo study, seabuckthorn seed oil inhibited the toxicity of CCl₄, as seen from the significantly increased activities of the antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. The GSH content in the liver was also increased, whereas hepatic malondialdehyde was reduced. Taken together, these results clearly indicate that seabuckthorn seed oil has significant potential as a natural antioxidant agent.     

Chemical composition and antimicrobial activity of the essential oil of Coriandrum sativum

The essential oil from leaves of Coriandrum sativum L. (Apiaceae), obtained by hydro-distillation was analysed by gas chromatography–mass spectrometry (GC–MS) and also evaluated for in vitro antimicrobial activity. Out of 27 peaks, 24 components, which constitute 92.7%, were identified in the oil. The oil was dominated by aldehydes and alcohols which accounted for 56.1% and 46.3% of the oil, respectively. The major constituents were 2E-decenal (15.9%), decanal (14.3%), 2E-decen-1-ol (14.2%) and n-decanol (13.6%). Other constituents present in fairly good amounts are 2E-tridecen-1-al (6.75%), 2E-dodecenal (6.23%), dodecanal (4.36%), undecanol (3.37%), and undecanal (3.23%). The oil was screened for antimicrobial activity against both Gram positive (Staphylococcus aureus, Bacillus spp.) and Gram negative (Escherichia coli, Salmonella typhi, Klebsiella pneumonia, Proteus mirabilis, Pseudomonas aeruginosae) bacteria and a pathogenic fungus, Candida albicans. The oil showed pronounced antibacterial and antifungal activity against all of the microbes tested, except for P. aeruginosae, which showed resistance.     

Changes on essential oil composition of coriander (Coriandrum sativum L.) fruits during three stages of maturity

The essential oils composition of coriander (Coriandrum sativum L.) fruits obtained by hydrodistillation was studied at three stages of maturity by GC–FID and GC–MS. Essential oil yields showed marked increase during maturation process and forty one compounds were identified. Geranyl acetate (46.27%), linalool (10.96%), nerol (1.53%) and neral (1.42%) were the main compounds at the first stage of maturity (immature fruits). At the middle stage, linalool (76.33%), cis-dihydrocarvone (3.21%) and geranyl acetate (2.85%) were reported as the main constituents. Essential oils at the final stage of maturity (mature fruits) consist mainly on linalool (87.54%) and cis-dihydrocarvone (2.36%). Additionally, accumulation of monoterpene alcohols and ketones was observed during maturation process of coriander fruit.     

In vitro and in vivo protein hydrolysis of beans (Phaseolus vulgaris) genetically modified to express different phaseolin types

Experiments were conducted to study whether phaseolin type could influence proteolysis susceptibility and nutritional value of total bean protein. The DOR-390 bean cultivar was genetically modified to express different phaseolin types (S, T or I). Beans were soaked and autoclaved. A sequential hydrolysis was carried out in vitro with pepsin and pancreatin. Differences in the degree of protein hydrolysis among bean lines started at 30 min and remained until 240 min, with the S bean proteins presenting lower values (P < 0.05). Subsequently, rats were fed with diets containing beans expressing different phaseolin types as the only source of protein for N digestibility and nutritional value determination. No differences (P > 0.05) in ileal protein digestibility and rat growth were observed. In conclusion, the differences in in vitro hydrolysis between bean lines expressing different phaseolin types had no consequences on growth and N retention in rats.     

Phenolic composition and inhibitory effect against oxidative DNA damage of cooked cowpeas as affected by simulated in vitro gastrointestinal digestion

Cowpeas contain phenolic compounds with potential health benefits. The effect of simulated gastrointestinal digestion on phenolic composition of cooked cowpeas and the ability of the digests to inhibit radical-induced DNA damage was determined. A red and a cream-coloured cowpea type were used. The phenolic composition of acetone extracts and enzyme digests of cooked cowpeas was determined using UPLC-MS. Compounds such as p-hydroxybenzoic acid, p-coumaric acid, coumaroylaldaric acid and feruloylaldaric acid were present in the acetone extracts of the cooked cowpeas but were not detected in the enzyme digests. Glycosides of quercetin and myricetin decreased upon in vitro gastrointestinal digestion of cooked cowpeas whereas flavan-3-ols were hardly present except catechin glucoside. The enzyme digest of the red cowpea type was about thrice as effective as that of the cream cowpea type in protecting DNA from oxidative damage. The observation that enzyme digests of cooked cowpeas inhibited radical-induced DNA damage suggests that cowpea phenolics retain some radical scavenging activity after gastrointestinal digestion.     

A comparative study of tocopherols composition and antioxidant properties of in vivo and in vitro ectomycorrhizal fungi

In aerobic organisms, the free radicals are constantly being produced during the normal cellular metabolism. The antioxidant properties of many organisms and particularly of wild mushrooms with their content in antioxidant compounds such as tocopherols, can detoxify potentially damaging forms of activated oxygen. Herein, a comparative study of tocopherols composition and antioxidant properties of in vivo (fruiting bodies) and in vitro (mycelia) ectomycorrhizal fungi: Paxillus involutus and Pisolithus arhizus. Tocopherols were determined by high performance liquid chromatography (HPLC) coupled to a fluorescence detector. The antioxidant properties were studied in terms of DPPH radical-scavenging activity, reducing power and inhibition of β-carotene bleaching. Fruiting bodies revealed the highest antioxidant properties, including scavenging effects on free radicals (EC₅₀ = 0.61 and 0.56 mg/ml) and inhibition of lipid peroxidation capacity (EC₅₀ = 0.40 and 0.24 mg/ml for P. involutus and P. arhizus, respectively), than mycelia produced in vitro cultures. Nevertheless, mycelia revealed higher levels of total tocopherols than fruiting bodies, and particularly P. arhizus mycelium proved to be a powerful source of γ-tocopherol (154.39 μg/g dry weight).     

Effect of soaking and hydrothermal processing methods on the levels of antinutrients and in vitro protein digestibility of Bauhinia purpurea L. seeds

The effects of various domestic processing methods such as soaking, cooking and autoclaving on the levels of certain antinutritional factors and in vitro protein digestibility of seeds of Bauhinia purpurea L., an underutilised legume collected from South India, were investigated. The raw seeds were found to contain antinutritional factors like total free phenolics (2.75 g/100 g), tannins (2.35 g/100 g), phytic acid (692 mg/100 g) and flatulence factors, raffinose (0.54 g/100 g), stachyose (1.17 g/100 g) and verbascose (0.95 g/100 g). Soaking the seeds in distilled water caused maximum reduction in the phytic acid content (37%), whereas soaking in NaHCO₃ solution reduced significant levels of phenolics and tannins (72% and 78%, respectively). A reduction in the levels of oligosaccharides (raffinose by 63%, stachyose by 42% and verbascose by 79%) was observed during cooking. Of the attempted treatments, autoclaving appeared to be most effective in reducing levels of all the investigated antinutrients, except phytic acid, and also improved the in vitro protein digestibility of B. purpurea seeds.     

Coupled extruder-headspace,a new method for analysis of the essential oil components of Coriandrum sativum fruits

A new method involving concurrent single screw extruder combined with continuous headspace dynamic for the extraction and identification of the essential oil of Coriandrum sativum L. fruit was developed. The effect of six different nozzle diameters (5, 6, 7, 8, 9 and 10 mm) on the content and chemical composition of the essential oil of coriander fruit was studied. The oils from fruit samples were obtained by OMEGA 20 extruder.     

Phenolic acid analysis and antioxidant activity assessment of oil palm (E. guineensis) fruit extracts

Phenolic compounds in oil palm fruit (E. guineensis) were extracted in soluble free (SFP), insoluble-bound (ISBP) and esterified (EFP) forms. The total phenolic content (TPC) of the oil palm fruit extracts was determined using the Folin–Ciocalteu method and found to range from 5.03 to 9.04 g/L per g of dried weight (DW). The antioxidant activities of oil palm phenolic extracts were analysed using free radical scavenging assays and results showed that oil palm phenolic extracts contained antioxidant activities in the order of ISBP > EFP > SFP. Eight different phenolic acids were identified and quantified using a simple reversed-phase high performance liquid chromatography (HPLC) with a diode array detector (DAD) and liquid chromatography/tandem mass spectrometry (LC/MS/MS). Ferulic, p-hydroxybenzoic and p-coumaric acid were the dominant phenolic acids found in oil palm fruit extracts and ranged from 55 to 376 μg/g of DW.     

Phenolic acids and flavonoids in leaf and floral stem of cultivated and wild Cynara cardunculus L. genotypes

Ten genotypes, cultivated and wild of Cynara cardunculus L. were evaluated for their content of phenolic acids, flavonoids and their antioxidant activity. The major compounds present in the leaf were luteolin derivatives in globe artichoke and apigenin derivatives in wild and cultivated cardoon. Apart from ‘Cimiciusa di Mazzarino’ (var. scolymus), caffeoylquinic acids represent the main phenolic compounds in the floral stem. In particular, ‘Sylvestris Creta’ (var. sylvestris) and ‘Violetto di Sicilia’ (var. scolymus) show the highest content of caffeoylquinic acid ∼95% of the total measured polyphenols. The antioxidant capacity, in both leaf and floral stem, was qualitatively and quantitatively dependent on the phenolic acid and flavonoid profile. The phenolic acids and flavonoids in normally uneaten parts of wild and cultivated artichoke could be exploited as sources of natural antioxidants.     

Quantification of the polyphenolic fraction and in vitro antioxidant and in vivo anti-hyperlipemic activities of Hibiscus sabdariffa aqueous extract

In the present study the quantification of the polyphenolic fraction, anthocyanins and other polar compounds, the antioxidant capacity and the anti-hyperlipemic action of the aqueous extract of Hibiscus sabdariffa has been achieved. Seventeen compounds were successfully quantified either by HPLC-DAD or HPLC-ESI-TOF-MS. Six of them were directly quantified by their corresponding standards, whereas the rest were indirectly quantified as equivalents using standards of similar compounds. The antioxidant capacity have also been estimated by comparing different assays, i,e, Trolox equivalent antioxidant capacity (TEAC), ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and measurement of thiobarbituric acid reacting substances (TBARS). H. sabdariffa showed high reducing capacity in FRAP assay and significant capability to scavenge peroxyl radicals in the ORAC assay. Nevertheless, the extract exhibited poor efficacy to inhibit peroxyl radicals in lipid systems. The plant extract also exhibited the capacity to decrease serum triglyceride concentration on hyperlipemic mouse model.     

The in vitro and in vivo effects of yerba mate (Ilex paraguariensis) extract on adipogenesis

The aim of this study was to evaluate the effects of yerba mate extract and its principal bioactive compounds on adipogenesis. The anti-adipogenic effects of yerba mate, chlorogenic acid, quercetin and rutin were evaluated in 3T3-L1 cells using a PCR array. The results obtained in vitro were validated in vivo in a high-fat diet-induced model of obesity. The in vitro and in vivo results demonstrated that yerba mate extract down-regulated the expression of genes that regulate adipogenesis, such as Creb-1and C/EBPα, and the extract up-regulated the expression of genes related to the inhibition of adipogenesis, including Dlk1, Gata2, Gata3, Klf2, Lrp5, Pparγ₂, Sfrp1, Tcf7l2, Wnt10b, and Wnt3a. In summary, it was demonstrated that yerba mate and its bioactive compounds regulate the expression of genes related to in vitro adipogenesis. Furthermore, yerba mate might regulate adipogenesis through the Wnt pathway.     

Characterisation of lipoxygenase from pea seeds (Pisum sativum var. Telephone L.)

Lipoxygenase (LOX) from pea seeds (Pisum sativum var. Telephone L.) was extracted and studied of biochemical properties. The molecular mass of purified lipoxygenase was 93 kDa. The effects of substrate specificity, pH, and sensibility to various inhibitors: caffeic acid, ferulic acid, benzoic acid, catechin, quercetin and kaempferol of LOX were investigated. Lipoxygenase showed the highest activity toward linoleic acid and the lowest toward oleic acid as substrates. Kinetic studies indicated that V_max of the LOX activity was 151.5 U/min and corresponding K_m value of 0.44 × 10⁻³ M. Optimum pH of lipoxygenase was reported at 5.5. Caffeic acid was the most effective inhibitor and kaempferol was the least effective.     

Antioxidant capacity of four polyphenol-rich Amazonian plant extracts: A correlation study using chemical and biological in vitro assays

Many plants used in Amazonian folk medicine present a high antioxidant activity. In this study, the antioxidant activities of four largely used plants, namely Byrsonima crassifolia, Davilla kunthii, Davilla rugosa and Inga edulis, were evaluated, using methanolic extracts of their leaves, fruits and bark and several different in vitro tests, based either on the capacity to scavenge free radicals (ORAC, TRAP) or on the ability to protect biological structures (LDLs, erythrocytes). The total phenolics (TP), flavanoids (TFA) and flavonols (TFO) were also measured. Almost all extracts performed well in all assays of antioxidative capacity, with best activities found in leaves (compared to fruits and bark). Most antioxidative performance indicators (ORAC, TRAP, LDL protection) correlated well with the TP and TFA content of the extracts. Conversely, correlation was lower between TFO and these indicators, reflecting a lower involvement of these compounds in antioxidant processes. Erythrocyte protection against oxidant-triggered haemolysis showed no correlation with any of the phenolic content indicators, suggesting that most of these compounds have a low ability to protect lipid targets in the erythrocyte membrane. On the other hand, protection of erythrocytes against haemolysis correlated positively with LDL protection. The extract of I. edulis leaves contained average amounts of polyphenols but ranked first in the majority of the tests, indicating the occurrence of particularly efficient compounds with very important antioxidant properties, which could be used for medicinal and other applications.     

Soymilk phenolic compounds,isoflavones and antioxidant activity as affected by in vitro gastrointestinal digestion

The aim of this research was to evaluate changes in the phenolic compounds, isoflavones and antioxidant activity of soymilk following in vitro gastrointestinal digestion (including dialysis). Gastric digestion significantly influenced the release of bioactive substances from the soymilk matrix, increasing the concentration of total phenolic components (35% as the sum of individuals and 14% by Folin–Ciocalteu [F–C] method), total isoflavone content (22%) and total antioxidant activity (76%). The concentration of all those compounds was reduced significantly in the duodenal fraction in comparison to gastric digestion and their lowest concentration was observed in the dialysed fraction, where phenolic acids were not detected. The bioaccessibility of soymilk phenolic compounds was 15% as the sum of individuals and 20% by F–C assay; isoflavones 36% and constituents with antioxidant activity 27%. Results suggest that most of these compounds were sufficiently available to be absorbed and could contribute health benefits.