Antioxidant activity and polyphenol content of Turkish thyme (Thymus vulgaris) monitored by liquid chromatography and tandem mass spectrometry

Like tea, the leaves of Turkish thyme (Thymus vulgaris) can be boiled in water to produce an extract. This is widely used as syrup for the treatment of coughs and bronchitis at alternative medicine clinics in many parts of the world. In the current study, we assessed the phenolic content and antioxidant activity of thyme. The antioxidant activities of both ethanol and aqueous extracts of thyme were determined using various in vitro methods. The total phenolic and total flavonoid contents were determined to be a gallic acid equivalent and a quercetin equivalent, respectively. Finally, the quantities of the phenolic compounds were detected using high-performance liquid chromatography and tandem mass spectrometry. The total phenolic compounds in the aqueous extract and ethanol extracts of Turkish thyme were 256.0 μg gallic acid equivalent/mg dried extract and 158.0 μg gallic acid equivalent/mg dried extract, respectively. Conversely, the total flavonoid compounds in both extracts were 44.2 μg and 36.6 μg quercetin equivalent/mg dried extract, respectively. For the first time, we determined phenolic contents and investigated the antioxidant potential of thyme. The results indicate that Turkish thyme is a good dietary source with phenolic properties.     

Protective effects of phenolic constituents from Cytisus multiflorus,Lamium album L. and Thymus citriodorus on liver cells

The present study investigated the antioxidant and cytoprotective effects of purified ethanolic extracts of Cytisus multiflorus, Lamium album L. and Thymus citriodorus plants. These extracts showed high antioxidant activity in DPPH and reducing power assays. Using a model of chemical stress induced by potassium dichromate (DK) in human hepatoblastoma HepG2 cells, 50 μg/mL of C. multiflorus, L. album and T. citriodorus extracts decreased the rate of reactive oxygen species (ROS) production by 35%, 26% and 20%, respectively, when exposed to 25 μM of DK. This effect was also observed for the treatment of cells with individual polyphenolic compounds determined in the extracts, or with mixtures prepared with individual polyphenolic compounds simulating the phenolic composition of the extracts. Additionally, the purified ethanolic extracts and the prepared polyphenolic mixtures showed a cytoprotective effect against DK-induced toxicity. The overall results emphasize the contribution of polyphenols in antioxidant and cytoprotective properties of the studied plants.     

Total phenolic content,antiradical, antioxidant and antimicrobial activities of Rhododendron honeys

Fifty Rhododendron honey samples obtained from Black Sea Region of Turkey were screened for total phenolic content by the modified Folin–Ciocalteu method, for potential antioxidant activity using phosphomolybdenum assay and by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) method for antiradical activity. The antimicrobial activity was studied by the agar diffusion method, using eleven bacteria and two yeasts.     

Phenolic compounds and abscisic acid as potential markers for the floral origin of two Polish unifloral honeys

The phenolic profiles of Polish honey samples from heather (Calluna vulgaris L.) and buckwheat (Fagopyrum esculentum L.) were determined. The phenolic components were isolated from honey samples using Amberlite-XAD-2 as a solid-phase extraction sorbent. For the determination of the composition of the honey extracts HPLC with photodiode array detector was applied. Identification and quantification of phenolics was performed by comparison of their retention times and UV spectra with those of standard solutions of pure reference substances and by using those references as external standards. From among 20 commercially available standards over 15 of them were identified as present in all analysed samples. Taking into account the samples of the same unifloral honeys, similar qualitative but slightly quantitatively different phenolic characteristic profiles were observed. These profiles might be considered as “fingerprints” of heather and buckwheat honeys.     

Antioxidant phenolic compounds from Smilax sebeana Miq.

Methanol extract and sub-fractions of Smilax sebeana rhizomes and roots were analyzed to evaluate the compounds involved in antioxidant activity. In all separated fractions of the different polarity solvents, ethyl acetate fraction showed the highest antioxidant activity and total phenolic content. This fraction was subjected to the sephadex LH-20 column and preparative HPLC for purification. Six phenolic compounds, chlorogenic acid (1), 4-formylphenol (2), epicatechin (3), cinchonain IIa (4), Ia (5) and Ib (6) were isolated and identified by spectroscopic analyses and further evaluated their potential antioxidant activities by DPPH and superoxide radical scavenging assays. Compared with synthetic antioxidant Trolox, except 4-formylphenol, the other isolated five compounds exhibited excellent antioxidant activities. This is the first report on the chemical constituents of S. sebeana which potentially involved in antioxidant activity. The results suggest that the ethyl acetate extract of S. sebeana might be used as a readily accessible source of natural antioxidant.     

Study of acrylamide mitigation in model system: Effect of pure phenolic compounds

Many natural antioxidant extracts were reported to potentially reduce the acrylamide formation during Maillard reaction. In this work, the effect of pure antioxidant molecules on acrylamide net amount was investigated in an equimolar asparagine–glucose system (10 mM) during heating (200 °C). All experimentations were performed in the newly designed test cell. Cinnamic acid and six phenolic compounds (gallic, ferulic, coumaric, caffeic acids, catechin, and epicatechin), displaying different antioxidant capacity, were added at 5 mM to the model system. The results indicated no reduction effect of the tested compounds on acrylamide net amount. Moreover, in the case of ferulic acid a slight but significant promoting effect was observed. An increase of antioxidant activity was also noticed during heating of model solution spiked with caffeic acid but the acrylamide level was not affected.     

Evaluation of the phenolic content,antioxidant activity and colour of Slovenian honey

Honey samples from the seven most common honey types in Slovenia were screened for total phenolic content by the modified Folin–Ciocalteu method, for potential antioxidant activity using the ferric reducing antioxidant power (FRAP) assay and by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method for antiradical activity. In addition the colour characteristics of honey samples were analysed. The results of the study showed that total phenolic content, antioxidant activity and colour parameters differ widely among different honey types. Phenolic content expressed as gallic acid equivalent ranged from 44.8 mg/kg in acacia honey to 241.4 mg/kg in fir honey. Antioxidant activity was the lowest in the brightest acacia and lime honeys and the highest in darker honeys, namely fir, spruce and forest. The colour of the Slovenian honeys, analysed in this study was very variable and ranged from pale yellow to dark brown. Correlations between the parameters analysed were found to be statistically significant (p < 0.05).     

Total phenolic contents and antioxidant activity of corn tassel extracts

Ground corn tassels, a by-product of corn, were used as a source of phenolic compounds. Water, ethanol, methanol, acetone, hexane, chloroform, butanol, petroleum ether and methylene chloride were evaluated as different polarity solvents to extract these phenolic compounds. Ethanol exhibited the highest extraction ability for such phenolic compounds, followed by methanol and water, where the total phenols were 0.1575%, 0.1125% and 0.0737%, respectively. Antioxidant activity of corn tassels ranged from 83.0% to 85.2%, 69.9% to 83.7%, 69.8% to 80.4%, 22.2% to 49.1% and 14.8% to 19.3% radical scavenging activity (% RSA) for ethanol, methanol, acetone, butanol and water extracts, respectively. The ethanolic extract of the corn tassels was successfully utilised to retard the oxidation of sunflower oil and the obtained induction period values were comparable to those of tert-butylhydroquinone (TBHQ).     

Determination of antioxidant activity and phenolic compounds of Muntingia calabura Linn. peel by HPLC‐DAD and UPLC‐ESI‐MS/MS

Antioxidant activity in Muntingia calabura Linn. peel was evaluated by DPPH radical, ORAC, ABTS cation radical, FRAP assays and total phenolic contents by different extraction conditions. In addition, a method for determination of phenolic compounds in calabura peel samples harvested in Brazil using methanol:water and magnetic stirring as the extraction method, HPLC‐DAD and UPLC‐ESI‐MS/MS analysis were developed. Calabura peel showed antioxidant activity for all extraction conditions and assays evaluated, the most polar solvents being more effective. The developed HPLC‐DAD method allowed the accurate determination of phenolic compounds, with recoveries in the range of 72–107% and precision values ≤4%, with exception for chlorogenic acid. Gallic acid was determined at the highest concentration levels, followed by myricetin, ferulic acid and vanillic acid. However, all the five proposed phenolic compounds were identified in calabura peel samples by UPLC‐ESI‐MS/MS. Thus, calabura peel, an uncommon edible fruit part, can be appointed as a rich source of phenolic compounds.     

Effects of various clarification treatments on phenolic compounds and color of apple juice

The purpose of this study was to investigate the changes that occur during conventional clarification using gelatin, bentonite, silica sol, and water-soluble chitosan on the phenolic compounds, antioxidant activity, and color of apple juice. The apple material used in this study was of two varieties: Sampion and Idared. The changes in the polyphenols composition (procyanidins, hydroxycinnamic derivatives, and dihydrochalcones) were monitored through the clarification process. Sampion apple control juices contained more total polyphenols than do Idared apple juices. In Sampion variety apple juice, the dominant polyphenols are the flavan-3-ols (86% of total polyphenols), followed by hydroxycinnamic acids (9.7%), dihydrochalcones (3.0%), and flavonols (1.3%). In Idared apple juice the hydroxycinnamic acids (especially chlorogenic acid) are dominant (about 48% of total polyphenols), followed by flavan-3-ols (40%). However, the concentration of polymeric procyanidins in Sampion apple juices was 62.8 and 46.3% less when the Profloc (chitosan) and gelatin treatments were used, respectively. Aktivbentonit and Puranit (bentonite) supplementary added in juices clarification have some protective effect on polymeric procyanidins only with Profloc treatment. That kind of effect was not observed in Idared apple juices with almost eight times smaller polymeric procyanidins concentration than in Sampion apple juices. The antioxidant activity, measured by the DPPH (1,1-diphenyl-2-picrylhydrazyl radical) method, ranged from 0.20 mg TEAC/mL in Idared apple juice to 0.30 mg TEAC/mL in Sampion apple juice, measured by the ABTS method, from 0.17 to 0.48 mg TEAC/mL, respectively. Clarification of apple juices with chosen clarifying agents has statistically no significant (p>0.05) influence on antioxidant capacity. This study suggests that chitosan can be used as a conventional clarifying aid of apple juices and that treatment has no impact on their biochemical parameters.     

Assessment of in vitro antioxidant capacity of the seed coat and the cotyledon of legumes in relation to their phenolic contents

Methanolic extracts of the seed coat and the cotyledon of two varieties of lentils (Lens culinaris L.) and two varieties of dark peas (Pisum sativum L.), were analysed for their antioxidant capacities (EC₅₀), in the form of free radical-scavenging activities. Huge differences have been observed in the antioxidant capacity in the seed coat and the cotyledon in both legumes. The seed coat, in which are located, principally, phenolic compounds with flavonoid structures, presents higher antioxidant activity than the cotyledon in lentils and peas, showing differences in both species but not very large differences between varieties. An analysis of principal components was applied to the results in order to relate the antioxidant capacity of these legumes with their phenolic composition previously established by the authors.     

Using flavonoids, phenolic compounds and headspace volatile profile for botanical authentication of lemon and orange honeys

This study evaluates the usefulness of flavonoids (naringenin, hesperetin, chrysin, galangin, kaempferol, luteolin, pinocembrin, and quercetin) and phenolic acids (caffeic acid and p-coumaric acid) together with 37 volatile compounds in the differentiation between lemon blossom honey (Citrus limon) and orange blossom honey (Citrus spp.). The total content of flavonoids and phenolic acids is twice as high in lemon honey (6.20 mg/100 g) as in orange honey (3.64 mg/100 g); naringenin and caffeic acid were the main compounds in all cases. Hesperetin, a floral marker of citrus honey, was not significantly different for the two types of honey. A multivariate PLS2 analysis showed that some volatile compounds such as, 4 lilac aldehydes and bencenacetaldehyde (all abundant in orange honey) were negatively correlated with 4 flavonoids: pinocembrin, chrysin, naringenin and quercetin, and caffeic phenolic acid (all abundant in lemon honey). Moreover, the last 5 compounds were positively correlated with: 6 alcohols, 2 ketones, acetaldehyde and furanmethanol. This is a first approach to employ all of these compounds together with appropriate statistical techniques to differentiate between two varieties of citrus honey, and therefore it could be an interesting tool for their authentication.     

Changes in antioxidant capacity,isoflavone profile,phenolic and vitamin contents in soymilk during extended fermentation

In this study, the influence of lactic acid bacteria fermentation on the antioxidant capacity, phenolic compounds and vitamins profile of soymilk with different final fermentation pH values (pH 4.55, 4.15 and 3.85) was examined. Fermented soymilk (FSM) with a final pH of 3.85 exhibited relatively higher antioxidant capacity than samples terminated at pH 4.55 or 4.15, as shown from results of Folin–Ciocalteu assay (57.3–63.8 mg GAE/100 mL FSM), DPPH-radical scavenging efficiency (30.0–36.5%) and ferricyanide reducing power (1.13–1.58 mg AAE/100 mL FSM). Among all products, Lactobacillus rhamnosus WQ2-fermented soymilk showed the highest antioxidant level among the four Lactobacillus species, while Lactobacillus acidophilus CSCC 2400 possessed the best ability to deglycosylate isoflavone glucoside (IG), reducing IG concentration by 60.8%. Four vitamins, three phenolic acids, one flavanol, and six isoflavones were detected in fermented and non-fermented soymilk by HPLC-DAD. The contents of antioxidant compounds in FSM were closely related to reducing power and anti-radical ability. In addition, antioxidant capacity was highly correlated with proteolytic activity. This study also demonstrated the potential of extended fermentation to enhance the overall health-promoting property of the products.     

Phenolic profile and antioxidant properties of Polish honeys

The phenolic components of honeys have great participation in their nutritional value and antioxidant activity. Moreover, phenolic components are promising markers for the determination of botanical and geographical origin of honey. The purpose of the present work was to study the antioxidant activity and profiles of phenolic acids and flavonoids of honeys of various origins. The total phenolic content of honeys varied from 4.46 to 15.04 mg of gallic acid equivalents per 100 g of product and the total phenolic acid content determined chromatographically varied from 201.05 to 2089.08 μg per 100 g of product. Buckwheat honey exhibited the highest antioxidant activity and contained the highest total phenolic amount, whereas rape honey exhibited the lowest values in this respect. Moreover, the buckwheat honey contained the highest amount of phenolic acids. There were significant linear correlations between total phenolic content and antioxidant activity of honey extracts in the reaction with DPPH^? (1,1‐diphenyl‐2‐picrylhydrazyl) and ABTS^{? +} (2,2′‐azinobis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid)) free radicals. In most samples, p‐coumaric acid was the dominant phenolic acid (39.1–677.2 μg per 100 g). The honeys also contained considerable amount of gallic acid (6.0–913.8 μg per 100 g). Among flavonoids naringenin was predominant in the most studied honey samples.     

The UPLC–ESI–QqQLIT–MS/MS method for quantitative determination of phytochemicals in ethanolic extracts of different parts of eight Ficus species: Development and validation

Ficus and validation of the ultra performance liquid chromatography–electrospray ionization hybrid triple quadrupole–linear ion trap–tandem mass spectrometry (UPLC–ESI–QqQ_LIT–MS/MS) method in a multiple-reaction monitoring (MRM) mode for the quantitative determination of 19 phytochemicals. The chromatographic separation of targeted phytochemicals was performed using the Waters ACQUITY UPLC BEH? C18 column (1.7 μm, 2.1 mm × 50 mm) with 0.1% formic acid with water and acetonitrile as a mobile phase at a flow rate of 0.25 mL/min. The validation parameters showed the overall recoveries from 95.78?101.44% (RSD ≤ 3.25%), precision (intra-day: RSD ≤ 2.96%; inter-day: RSD ≤ 2.89%), linearity (R² ≥ 0.9982), limit of detection (8.60 × 10^–10?2.18 × 10^–6 mg/mL), and the limit of quantitation (2.60 × 10^–9–6.63 × 10^–6 mg/mL) in the concentration range from 0.5 to 1000 × 10^–6 mg/mL. This method was successfully applied in ethanolic extracts of different parts (fruits, leaves, and barks) of selected eight Ficus species. Quinic acid was predominant followed by rutin and chlorogenic acid among the studied nineteen phytochemicals. Ficus benjamina showed the maximum total content in fruits and leaves. The UPLC–ESI–QqQ_LIT–MS/MS method combined with principal component analysis (PCA) was successfully used for Ficus species discrimination on the basis of the contents of 15 compounds. The UPLC–ESI–QqQ_LIT–MS/MS method combined with PCA could be used for quality control.     

Changes of phenolic acids and antioxidant activities during potherb mustard (Brassica juncea, Coss.) pickling

Phenolic acids in potherb mustard (Brassica juncea, Coss.) were determined and the effects of pickling methods on the contents of total free phenolic acids, total phenolic acids, total phenolics, and antioxidant activities were investigated. Gallic acid, protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, caffeic acid, p-coumaric acid, ferulic acid, and sinapic acid were identified in the present study. The contents of total free phenolic acids, total phenolic acids and total phenolics in fresh potherb mustard were 84.8 ± 0.58 μg/g dry weight (DW), 539 ± 1.36 μg/g DW, and 7.95 ± 0.28 mg/g DW, respectively. The total free phenolic acids increased during the pickling processes, but the total phenolic acids, total phenolics, and antioxidant activities decreased. However, after 5 weeks of fermentation, all the pickling methods retained over 70% of total phenolic contents and above 65% of antioxidant capacities. The results indicated that pickling processes were relatively good methods for the preservation of phenolic acids and antioxidants for potherb mustard.     

Volatile and carbohydrate composition of rare unifloral honeys from Spain

Volatile composition and carbohydrate content of Spanish honey samples from uncommon botanical origins have been studied by gas chromatography coupled to mass spectrometry. About 100 volatile compounds were identified; some of them appeared to be characteristic of particular honey types, such as methyl salycilate in willow (Salix spp.), 2,6,6-trimethyl-2,4-cycloheptadien-1-one (eucarvone) in almond tree (Prunus dulcis) and isophorone in strawberry-tree (Arbutus unedo). Concentration ranges for major carbohydrates were similar to those previously reported in other honeys with different botanical origins, although concentrations of maltulose in avocado honeys (Persea americana) and of melezitose in Quercus ilex honeys were higher. Some carbohydrate alcohols could also be considered as markers of honey botanical origin, such as quercitol for Q. ilex and perseitol for avocado.     

Variation in contents of phenolic compounds during growth and post-harvest storage of pigeon pea seedlings

The variation in contents of seven phenolic compounds in pigeon pea seedlings during growth and storage was investigated. Maximum contents of vitexin, isovitexin and orientin were found in leaves growing 40 days, which were 0.99 ± 0.06, 6.63 ± 0.35 and 30.89 ± 1.92 mg/g DW. Apigenin and luteolin were extensively distributed in leaves, stems and roots. Pinostrobin and cajaninstilbene acid were mainly accumulated in leaves, the peak values 3.53 ± 0.18 and 2.49 ± 0.13 mg/g DW appeared at the 60th day. Slight and steady increases of seven phenolic compounds were found in room temperature (25 °C) stored pigeon pea leaves up to 120 days. The highest accumulation of seven phenolic compounds at chilling temperature (4 °C) was observed at the 45th day, after which the contents decreased sharply. The stems extracts exhibited more efficient DPPH radical-scavenging ability while the roots extracts demonstrated the strongest lipid peroxidation inhibitory activity.     

Identification,characterisation, and quantification of phenolic compounds in the antioxidant activity-containing fraction from the seeds of Korean perilla (Perilla frutescens) cultivars

The present research was the first to investigate phenolic compound profiles and antioxidant properties in the seeds of various perilla (Perilla frutescens) cultivars. The 80% methanol extract (50 μg/ml) of this species showed potent antioxidant activities against 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals. Phenolic compounds were characterised by nuclear magnetic resonance (NMR) spectroscopy, and ultra performance liquid chromatography with photodiode array detector and electrospray ionisation/mass (UPLC-PDA-ESI/MS) analysis. Nine compounds were elucidated as caffeic acid-3-O-glucoside (1), caffeic acid (2), luteolin-7-O-glucoside (3), apigenin-7-O-glucoside (4), rosmarinic acid-3-O-glucoside (5), rosmarinic acid (6), luteolin (7), apigenin (8), and chrysoeriol (9). The individual and total phenolic contents were remarkably different, especially rosmarinic acid-3-O-glucoside (5) and rosmarinic acid (6) which were the predominant compounds (>95%) in all perilla cultivars. Additionally, Yeupsil cultivar exhibited the highest phenolic content (5029.0 μg/g) and antioxidant activity, whereas the lowest was shown by Dasil (2138.7 μg/g). Therefore, these results suggest that antioxidant effects of perilla seeds are correlated with phenolic contents.     

Egyptian chia seeds (Salvia hispanica L.) during germination: Upgrading of phenolic profile,antioxidant, antibacterial properties and relevant enzymes activities

Food Science and Biotechnology - Little studies on chia sprouts were not deeply address the polyphenols profiles and their functional properties during long period of germination. This study aims...