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Secondary metabolite content in rhizomes, callus cultures and in vitro regenerated plantlets of Solidago chilensis.

An in vitro culture system leading to the formation of callus and plant regeneration, starting from nodal sections and shoot tips, was developed for Solidago chilensis (Asteraceae). The content of the gastroprotective diterpene solidagenone as well as the phenolics chlorogenic acid (CA) and rutin was determined either in rhizomes from wild growing plants and in callus and in in vitro regenerated plantlets by analytical HPLC. Additionally, total phenolic and flavonoid content was assessed in plant samples, callus and cell suspensions. In terms of dry starting material, the percentual solidagenone content in nine S. chilensis samples ranged from 0.5-3.5% for rhizomes from wild growing plants, 0.1-0.3% for callus and 0.3% for an in vitro regenerated plantlet, respectively. The highest solidagenone contents were found in the wild plant during the late summer in the months of March and April (3.5-2.2%) while highest values for chlorogenic acid (0.5%) and rutin (0.4%) were detected in May, before senescence. The callus tissue and cell suspensions contained some 1.8-2.0 and 1.2% of total phenolics, respectively. CA was the main phenolic in the cell suspension while only traces were found in the callus. Rutin was not detected in the callus nor cell culture.

Cells, Cultured↗

Influence of cooking process on phenolic marker compounds of vegetables.

Phenolic compounds are secondary plant metabolites which have long been associated with flavor and color characteristics of fruits and vegetables. These phenolic compounds attract great interest due to their postulated health protecting properties. However, adequate intakes and absorption rate of phenolic compounds are necessary for these beneficial effects. Until now, little is known about alterations of phenolic compounds content by the cooking process. In the present study, the influence of different volumes of cooking water on the amount of selected phenolic marker compounds resting in the vegetables was assessed. In zucchini, rutin was quantified as a marker for flavonoid glycosides. Chlorogenic acid, representative of phenolic acids was analyzed in carrots. In beans, rutin and quercitrin, both belonging to flavonoid glycosides, were investigated. In potatoes, chlorogenic and caffeic acid were determined. The cooking of zucchini, beans and carrots with smaller amounts of water resulted in significant higher content of phenolic phytochemicals in the vegetables compared to cooking with larger water volumes. For potatoes, which showed great variations in content of phenolic acids after cooking, no significant differences in phenolic acids was observed. It can be concluded from these observations, that real intakes of phenolic compounds from cooked vegetables are lower and that the amounts consumed are therefore overestimated.

Caffeic Acids↗

Effects of plant-derived phenols on rat liver cytochrome P450 2B1 activity.

Dietary constituents contain a variety of compounds that are known to modulate liver enzyme activity. In this report, the plant-derived phenols catechin, chlorogenic acid, diosmin, epigallo-catechin gallate (EGCG), naringenin, quercetin and resveratrol were studied for their effects on the activity of cytochrome P450 2B1 in liver microsomes from 6- and 20-month male Fisher F344 rats. The compounds at two concentrations (0.1 and 0.25 mM) were incubated with 0.2 mg liver microsomal protein and 50 microM 7-ethoxy-4-trifluoromethyl coumarin (EFC). O-deethylation of EFC to the fluorescence product 7-hydroxy-4-trifluoromethyl coumarin (HFC) is catalyzed by CYP450 2B1. EGCG, naringenin, quercetin and resveratrol inhibited the in vitro O-deethylation of EFC in liver microsomes from both 6- and 20-month rats. Quercetin was the most effective inhibitor. Catechin inhibited the in vitro O-deethylation of EFC only in microsomes from 6-month-old rats whereas diosmin inhibited the reaction only in microsomes from 20-month-old rats. Chlorogenic acid inhibited the in vitro O-deethylation of EFC in microsomes from both age groups at the 0.25 mM concentration only. These results suggest that plant phenols have varied effects on liver microsomal cytochrome P450 2B1 activity that may be influenced by the age of the animal.

Alkylation↗

Potato peel extract-a natural antioxidant for retarding lipid peroxidation in radiation processed lamb meat.

The effective utilization of potato peel, a waste generated in large quantities by the food industry, as an antioxidant was investigated. Potato peel extract (PPE) exhibited high phenolic content (70.82 mg of catechin equivalent/100 g), chlorogenic acid (27.56 mg/100 g of sample) being the major component. The yield of total phenolics and chlorogenic acid increased by 26 and 60%, respectively, when the extract was prepared from gamma irradiated (150 Gy) potatoes. PPE showed excellent antioxidant activity as determined by beta-carotene bleaching and radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH). The suitability of PPE for controlling lipid oxidation of radiation processed lamb meat was also investigated. PPE (0.04%) when added to meat before radiation processing was found to retard lipid peroxidation of irradiated meat as measured by TBA number and carbonyl content. The antioxidant activity of PPE was found to be comparable to butylated hydroxytoluene (BHT).

Animals↗

4-Hydroxycinnamic Acid Hydroxylase and Polyphenolase Activities in Sorghum vulgare.

Green shoots and first internodes of Sorghum vulgare var. Wheatland milo contain three phenoloxidase activities separable by means of Sephadex G-100 gel filtration. Two of these are found only in green leaves. I, eluted at the void volume and presumably a high molecular weight form, has both monophenol and diphenol oxidase functions; II, an intermediate molecular weight form displays only a diphenol function; III, a low molecular form found only in first internodes, catalyzes the hydroxylation of 4-hydroxycinnamic acid to caffeic acid and may have a weak diphenol activity. The hydroxylase activities of peaks I and III were completely inhibited by boiling or by 1 millimolar diethyldithiocarbamate and were partially inhibited by 1 millimolar KCN. The time courses of the two monophenol activities differ in that the activity of internode tissue was linear for at least 3 hours while that of the leaf began to decrease after 15 minutes. Both O(2) and a suitable electron donor were obligatory. At pH 6, ascorbic acid and 2-amino-4-hydroxy-6, 7-dimethylpteridine were the best electron donors, while NADPH was less effective. The diphenol oxidase functions of forms I and II in leaf preparations were not identical. The activity of I was less stable than that of II. While both were more active with chlorogenic acid, the ratio of activity with chlorogenic acid to that with 3,4-dihydroxyphenylalanine was less than 50 for I and greater than 50 for II.

Journal Article↗

Anti-human immunodeficiency virus activity of 3,4,5-tricaffeoylquinic acid in cultured cells of lettuce leaves.

3,4,5-Tricaffeoylquinic acid (TCQA) that is not found in intact plant of lettuce leaves was isolated from the cultured cells. The intact plant produced chicoric acid (dicaffeoyl tartaric acid: L-CCA) as well as chlorogenic acid (3-caffeoylquinic acid: 3-CQA) as the major metabolites. After subculturing of the cells for 40 days, the amount of 3,4,5-TCQA reached to 0.14 mg/g fresh weight. The inhibitory effect of 3,4,5-TCQA for human immunodeficiency virus (HIV) Type 1 integrase was assayed. Anti-HIV activity using HIV and MT-2 cells was 1.15 microM and IC(50) against HIV integrase was 0.063 microM whereas cell toxicity of this chemical was expressed as 5% death of all living cells to be 18.4 microM. The HIV inhibitory effect of 3,4,5-TCQA was the highest in values among L-CCA, and other dicaffeoylquinic acids. This data will provide a new possibility for creating a new drug design for HIV.

Anti-HIV Agents↗

Antimutagenic activity of some naturally occurring compounds towards cigarette-smoke condensate and benzo[a]pyrene in the Salmonella/microsome assay.

Several compounds, occurring in food, were tested for antimutagenic activity towards cigarette-smoke condensate (CSC) and benzo[a]pyrene (BaP). Antimutagenicity was determined in the Salmonella/microsome test, with tester strain TA98, in the presence of rat-liver homogenate. Dose-response curves did show reduction of CSC- and BaP-induced mutagenicity by ellagic acid, riboflavin and chlorophyllin. Chlorophyll a and chlorophyll b, although less distinct, also inhibited CSC- and BaP-induced mutagenicity. Ascorbic acid, beta-carotene, tocopherol acetate, chlorogenic acid and butyl hydroxyanisole did not have any influence on the mutagenicity of CSC and BaP. The similarity in results for cigarette-smoke condensate and for BaP indicates that a general mechanism may be involved in the inhibition of CSC- and BaP-induced mutagenicity.

Animals↗

Bioactive phenolics from Carthamus lanatus L.

Two flavonoid aglycons, eight flavonoid glycosides, chlorogenic acid and syringin were isolated from aerial parts of Carthamus lanatus. Isorhamnetin 3-O-beta-D-glucoside and chlorogenic acid were found for the first time in the genus Carthamus and respectively, quercimeritrin, astragalin, kaempferol 3-O-beta-D-sophoroside and syringin in the species. The ethyl acetate fraction of the methanol extract exhibited a higher antioxidant activity than the butanol fraction measured by the alpha,alpha-diphenyl-beta-picrazylhydrazyl (DPPH) free radical scavenging assay. Cytotoxicity and antioxidant activities of the main constituent, luteolin 7-O-beta-D-glucoside, were evaluated.

Animals↗

Improved HPLC determination of phenolic compounds in cv. golden delicious apples using a monolithic column.

A rapid HPLC-DAD determination of phenols in apple using an RP monolithic column is reported. Because of the hydrodynamic advantages offered by this kind of column and the use of acidified acetonitrile as eluent, assays of apple extracts can be performed in <21 min. Assays of pulp and peel extracts were carried out without the need for time-consuming sample pretreatment except filtration. Several flavanols, hydroxycinnamic acids, dihydrochalcones, and six quercetin glycosides were identified and quantified. A seventh quercetin derivative, two chalcone-related compounds, and three hydroxycinnamic derivatives were also found. Peels proved to be richer in phenols than pulps, the former being composed mainly of (-)-epicatechin, procyanidin B2, chlorogenic acid, phloridzin, hyperin, and avicularin. In pulps, where the chlorogenic acid was the principal phenolic compound, quercetin glycosides were found in very low amounts.

Acetonitriles↗

Quality of organically and conventionally grown potatoes: four-year study of micronutrients, metals, secondary metabolites, enzymic browning and organoleptic properties.

The quality of potatoes from organic and conventional farming was investigated in this study. Tubers of eight potato varieties, organically and conventionally produced at one or two geographical sites in controlled field trials, were collected in four consecutive harvests from 1996-1999. The parameters analysed included nitrate, trace elements (As, Cd, Co, Cu, Fe, Hg, Mn, Ni, Pb, Se, Zn), vitamin C, potato glycoalkaloids, as well as chlorogenic acid, polyphenol oxidase and rate of tuber enzymic browning. The results indicated lower nitrate content and higher vitamin C and chlorogenic acid content to be the parameters most consistently differentiating organically from conventionally produced potatoes. Elevated concentrations of glycoalkaloids were also observed throughout the experiments in some potato varieties grown in organic farming systems. Principal component analysis (PCA) of the analytical and other data using three PCs confirmed a good separation between the organically and conventionally produced potatoes when studied in single crop years. However, score-plots (objects) and loading-plots (variables) of pooled results from the consecutive harvests showed that between the years' changes and also variety as well as geographical variations are equally or more important factors determining the quality of potatoes than the farming system. Further studies of various marker compounds of potato quality related to the organic or conventional farming systems should be performed before unbiased information can be given to the consumers.

Alkaloids↗

[Studies on chemical constituents from Eucommia ulmoides Oliv].

OBJECTIVE: To study the chemical constituents from Eucommia ulmoides Oliv. METHODS: The chemical constituents were isolated and repeatedly purified on silica gel column. They were identified and structurally elucidated by means of physio-chemical constants and spectral analysis. RESULTS: Seven compounds were obtained. They were (-)epicatechin (I), catechin (II), n-octacosanoic acid (III), tetracosanoic-2, 3-dihydroxypropylester (IV), rutin (V), chlorogenic acid (VI), and caffeic acid (VII), respectively. CONCLUSION: Four compounds (II, III, IV, V) were isolated from bark of Eucommia ulmoides Oliv for the first time.

Caffeic Acids↗

Chemical fingerprinting of valeriana species: simultaneous determination of valerenic acids, flavonoids, and phenylpropanoids using liquid chromatography with ultraviolet detection.

The roots and rhizomes of various valeriana species are currently used as a sleeping aid or mild sedative. A liquid chromatography method has been developed that permits the analysis of chlorogenic acid, lignans, flavonoids, valerenic acids, and valpotrates in various valerian samples. The best results were obtained with a Phenomenex Luna C18(2) column using gradient elution with a mobile phase consisting of water and 0.05% phosphoric acid and 2-100% acetonitrile-methanol (1 + 1) with 0.05% phosphoric acid. The flow rate was 0.8 mL/min and ultraviolet detection was at 207, 225, 254, 280, and 325 nm. Different valerian species and commercial products showed remarkable quantitative variations. Chlorogenic acid (0.2-1.2%), 3 lignans, linarin (0.002-0.24%), and valepotriates were detected in all the valeriana species analyzed. Highest amounts of valerenic acids were detected in V. officinalis L., trace amounts in V. sitchensis, and none in the other species analyzed.

Acetonitriles↗

[Studies on enzymic browning of potatoes (Solanum tuberosum). II. The quantitative relationship between browning and its causative factors (author's transl)].

Ten potato varieties, with different rates of browning, were analyzed quantitatively for phenoloxidase, tyrosine, chlorogenic acid, caffeic acid, and for reducing substances (ascorbic acid). The rate of tyrosine turnover was calculated from the data. The fact that the further reactions of the primary oxidation products leading to browning only take place after complete oxidation of the reducing substances, was taken into account. This leads to the same classification of the varieties as does visual observation of the rate of discolouration. Thus a clear relationship between browning and potato constituents is demonstrated.

Ascorbic Acid↗

Inhibitory effects of an aqueous extract of Apocynum venetum leaves and its constituents on Cu(2+)-induced oxidative modification of low density lipoprotein.

An aqueous extract of Apocynum venetum leaves and its constituents inhibited thiobarbituric acid reactive substances (TBARS) and conjugated-diene formation in the Cu(2+)-induced oxidation of low density lipoprotein (LDL) in vitro. The TBARS formation was most strongly inhibited by chlorogenic acid with an IC(50) value of 1.9 microM, but other constituents were in a range of 2.3-23.3 microM. On the other hand, the lag time in the conjugated-diene formation was dose-dependently prolonged by addition of the aqueous extract. Catechin prolonged the lag time more than 300 min and other constituents such as chlorogenic acid, epicatechin, epigallocatechin, hyperoside and isoquercitrin led to no conjugated-diene formation within 700 min under the experimental conditions.

Cholesterol, LDL↗

Responses of secondary chemicals in sugar maple (Acer saccharum) seedlings to UV-B, springtime warming and nitrogen additions.

Anticipated effects of climate change involve complex interactions in the field. To assess the effects of springtime warming, ambient ultraviolet-B radiation (UV-B) and nitrogen fertilization on the foliar chemistry and herbivore activity of native sugar maple (Acer saccharum Marsh.) seedlings, we carried out a field experiment for 2 years at two sugar maple forests growing on soils of contrasting acidity. At the Oliver site, soils are derived from a strongly calcareous till, whereas the naturally acidic soils and base-poor soils of the Haliburton site are derived from the largely granitic Precambrian Shield. At both sites, removal of ambient UV-B led to increases in chlorogenic acid and some flavonoids and reduced herbivore activity. At Haliburton, ammonium nitrate fertilization led to further increases in foliar manganese (Mn), whereas at Oliver there were no such changes. Nitrogen additions led to decreases in the concentrations of some flavonoids at both sites, but seedlings at Oliver had significantly higher concentrations of flavonoids and chlorogenic acid than seedlings at Haliburton. We suggest that this could be associated with increased mobilization of Mn due to increased soil acidity, which interferes with the role of calcium (Ca) in the phenolic biosynthetic pathway. It appears that the composition of the forest soil governs the response of seedlings when they are exposed to abiotic stressors.

Acer↗

Active components from Artemisia iwayomogi displaying ONOO(-) scavenging activity.

Peroxynitrite (ONOO(-)) is one of cytotoxic species produced by the reaction between superoxide (*O(2) (-)) and nitric oxide (NO). The main aim of this study was to identify ONOO(-) scavenging constituents from herbs. Methanolic extracts derived from one hundred thirty eight herbs were screened out for their ONOO(-) scavenging activities. Twenty herbs showed excellent ONOO(-) scavenging activity in a dose dependent manner. Of them, Artemisia iwayomogi, which showed the highest activity, was fractioned with several solvents. The ONOO(-) scavenging activity of fractions was in order of ethyl acetate (EtOAc) > n-butanol (BuOH) > dichloromethane (CH(2)Cl(2)) > water (H(2)O) fraction. The EtOAc and the n-BuOH fractions were further purified to isolate active principles by column chromatography. Apigenin 7-methylether (genkwanin), apigenin 7,4'-di-O methylether, jaceosidin, and scopoletin from the EtOAc fraction and chlorogenic acid, 2,4-dihydroxy 6-methoxy acetophenone 4-O-beta-D-glucoside, quebrachitol, and scopolin from the n-BuOH fraction were identified. The scavenging activities of chlorogenic acid (IC(50) = 0.52 +/- 0.04 microM), genkwanin (IC(50) = 1.01 +/- 0.10 microM), and scopoletin (IC(50) = 1.03 +/- 0.15 micro M) were higher than or comparable to a well-known ONOO(-) scavenger, penicillamine (IC(50) = 1.76 +/- 0.18 microM), suggesting that those compounds might be developed as effective ONOO(-) scavengers for, prevention of the ONOO(-)-involved diseases.

Artemisia↗

Plant phenolics as dietary antioxidants for herbivorous insects: a test with genetically modified tobacco.

High foliar phenolics are generally assumed to increase resistance to insect herbivores, but recent studies show that tobacco lines modified to over- and underexpress phenolics do not exhibit higher constitutive resistance to caterpillars. This is contrary to the expectation that ingestion of tobacco phenolics, particularly chlorogenic acid, should cause oxidative stress in herbivores. We investigated free radical production and antioxidant capacity of fresh crushed leaves of tobacco lines exhibiting over a sixfold difference in chlorogenic acid content to test whether high phenolic concentrations are associated with increased production of reactive oxygen species (ROS). The effects of in planta phenolic levels on feeding behavior, growth, biochemical markers of oxidative stress, and the antioxidant capacity of midgut fluid and hemolymph were assessed in tobacco budworm, Heliothis virescens. The experiments showed that high phenolic foliage was more prooxidant than low phenolic foliage, but the net balance in crushed tissue was antioxidant in comparison to buffer and the commercial antioxidant standard, Trolox. In H. virescens, the antioxidant capacity of midgut fluid was also powerful, and caterpillars fed high phenolic foliage did not exhibit the expected markers of oxidative stress in midgut tissues (altered ascorbate ratios, disulfides, or total hydroperoxides). Instead, hemolymph of larvae fed high phenolic foliage exhibited improved total Trolox equivalent antioxidant capacity (TEAC). These results suggest that the elevated foliar phenolics in some plants may have beneficial antioxidant properties for herbivorous insects, much as dietary phenolics do in mammals.

Adaptation, Physiological↗

Chromatographic performance of a new polar poly(ethylene glycol) bonded phase for the phytochemical analysis of Hypericum perforatum L.

The aim of this study was to evaluate the chromatographic performance of a poly(ethylene glycol) (PEG) stationary phase for the HPLC analysis of the secondary metabolites (chlorogenic acid, flavonoids, phloroglucinols and naphthodianthrones) in methanolic extracts of Hypericum perforatum L. (St. John's Wort) flowering tops, herbal medicinal products and dietary supplements. A fast and reliable method was developed. The analyses were carried out on a Supelco Discovery HS PEG column (150 mm x 4.6 mm i.d., 5 microm). A gradient mobile phase, composed of 0.1 M aqueous acetic acid solution (pH 2.8) and methanol-acetonitrile (5:4, v/v), was used. The flow rate was 1 mL/min. The photodiode array detector monitored the eluent at 270 (for chlorogenic acid, flavonoids and phloroglucinols) and 590 nm (for naphthodianthrones). The column was maintained at room temperature. The total running time was 40 min. The method was validated and showed good linearity, precision, accuracy, sensitivity and specificity. Through the above described phytochemical markers, this technique allowed the unequivocal identification and standardization of H. perforatum plant material and phytoproducts. The quantification data highlighted the fact that the products on sale, in particular those labeled as dietary supplements, varied widely in the quantitative composition of the active constituents. The developed method could be considered suitable for the quality control of H. perforatum herb and derivatives.

Chromatography, High Pressure Liquid↗