Redox properties of individual quercetin moieties.

Quercetin is one of the most prominent and widely studied flavonoids. Its oxidation has been previously investigated only indirectly by comparative analyses of structurally analogous compounds, e.g. dihydroquercetin (taxifolin). To provide direct evidence about the mechanism of quercetin oxidation, we employed selective alkylation procedures for the step-by-step blocking of individual redox active sites, i.e. the catechol, resorcinol and enol C-3 hydroxyls, as represented by newly prepared quercetin derivatives 1-3. Based on the structure-activity relationship (SAR), electrochemical, and computational (density functional theory) studies, we can clearly confirm that quercetin is oxidized in the following steps: the catechol moiety is oxidized first, forming the benzofuranone derivative via intramolecular rearrangement mechanism; therefore the quercetin C-3 hydroxy group cannot be involved in further oxidation reactions or other biochemical processes. The benzofuranone is oxidized subsequently, followed by oxidation of the resorcinol motif to complete the electrochemical cascade of reactions. Derivatization of individual quercetin hydroxyls has a significant effect on its redox behavior, and, importantly, on its antiradical and stability properties, as shown in DPPH/ABTS radical scavenging assays and UV-Vis spectrophotometry, respectively. The SAR data reported here are instrumental for future studies on the oxidation of biologically or technologically important flavonoids and other polyphenols or polyhydroxy substituted aromatics. This is the first complete and direct study mapping redox properties of individual moieties in quercetin structure.

Paramecium caudatum as a source of nitric oxide: Chemiluminescent detection based on Bluestar® Forensic reagent connected with microdialysis.

Nitric oxide (NO) chemistry inside the body is the most interesting part of its behavior. NO is involved in controlling blood pressure, and in transmitting nerve signals and a variety of other signaling processes. To explain the behavior of NO, it is necessary to determine its immediate concentration or observe time-dependent changes in its concentration. In Paramecium caudatum, NO is formed by calcium-dependent nNOS (NOS1)-like protein, which is distributed in the cytoplasm. NO synthesis affects the ciliary beat and consequent motility of cells and blocked NO synthesis reduces the ability of cells to move. The possibility of online coupling of microdialysis (of P. caudatum solution) with NO detection is demonstrated. Direct measurement of NO is carried out using dilute Bluestar® Forensic reagent (luminol-H2 O2 system; one of the NO detections is based upon the chemiluminescent reaction between NO and the luminol-H2 O2 system, which is specifically reactive to NO). The effect of a nitric oxide synthase inhibitor, NG-nitro-l-arginine methyl ester was observed. NO production was inhibited and the movement of P. caudatum was restricted. These effects were time dependent and after a specific time were reversed.

Lipidomic analysis of plasma, erythrocytes and lipoprotein fractions of cardiovascular disease patients using UHPLC/MS, MALDI-MS and multivariate data analysis.

Differences among lipidomic profiles of healthy volunteers, obese people and three groups of cardiovascular disease (CVD) patients are investigated with the goal to differentiate individual groups based on the multivariate data analysis (MDA) of lipidomic data from plasma, erythrocytes and lipoprotein fractions of more than 50 subjects. Hydrophilic interaction liquid chromatography on ultrahigh-performance liquid chromatography (HILIC-UHPLC) column coupled with electrospray ionization mass spectrometry (ESI-MS) is used for the quantitation of four classes of polar lipids (phosphatidylethanolamines, phosphatidylcholines, sphingomyelins and lysophosphatidylcholines), normal-phase UHPLC-atmospheric pressure chemical ionization MS (NP-UHPLC/APCI-MS) is applied for the quantitation of five classes of nonpolar lipids (cholesteryl esters, triacylglycerols, sterols, 1,3-diacylglycerols and 1,2-diacylglycerols) and the potential of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is tested for the fast screening of all lipids without a chromatographic separation. Obtained results are processed by unsupervised (principal component analysis) and supervised (orthogonal partial least squares) MDA approaches to highlight the largest differences among individual groups and to identify lipid molecules with the highest impact on the group differentiation.

Isoquercitrin: pharmacology, toxicology, and metabolism.

The flavonoid isoquercitrin (quercetin-3-O-ß-d-glucopyranoside) is commonly found in medicinal herbs, fruits, vegetables and plant-derived foods and beverages. This article reviews the occurrence, preparation, bioavailability, pharmacokinetics, toxicology and biological activity of isoquercitrin and "enzymatically modified (α-glucosylated) isoquercitrin" (EMIQ). Pure isoquercitrin can now be obtained on a large scale by enzymatic rutin hydrolysis with α-l-rhamnosidase. Isoquercitrin has higher bioavailability than quercetin and displays a number of chemoprotective effects both in vitro and in vivo, against oxidative stress, cancer, cardiovascular disorders, diabetes and allergic reactions. Although small amounts of intact isoquercitrin can be found in plasma and tissues after oral application, it is extensively metabolized in the intestine and the liver. Biotransformation of isoquercitrin includes deglycosylation, followed by formation of conjugated and methylated derivatives of quercetin or degradation to phenolic acids and carbon dioxide. The acceptable daily intake of (95%) isoquercitrin and of EMIQ was estimated to be 5.4 and 4.9mg/kg/day, respectively. Adverse effects of higher doses in rats included mostly (benign) chromaturia; nevertheless some drug interactions may occur due to the modulation of the activity and/or expression of drug metabolizing/transporting systems. With respect to the safety, affordability and beneficial pharmacological activities, highly pure isoquercitrin is a prospective substance for food supplementation.

Black and green tea--how to make a perfect crime.

The antioxidant properties of the black and green tea are well known. The latent bloodstains are detectable by luminol. The bloodstains also can be cover up by drinks and foods containing the antioxidants; thus their presence can cause a decrease of the luminol light emission (false-negative results). The aim of this study was to quantify the light emission decrease of the chemiluminescent mixture prepared according to Weber (containing NaOH) and the chemiluminescent mixture of pH 7.4 (for the determination of the total antioxidant capacity) for the open air-dried sample. The black and green teas and white wine were used as the antioxidant's samples (high and low total antioxidant capacity). The significant decrease of the luminol chemiluminescent emission caused by the presence of the black and green teas (and comparable for both of them) was observed in comparison with the presence of white wine.

Electrochemical sensing of total antioxidant capacity and polyphenol content in wine samples using amperometry online-coupled with microdialysis.

This work describes the method for total antioxidant capacity (TAC) and/or total content of phenolics (TCP) analysis in wines using microdialysis online-coupled with amperometric detection using a carbon microfiber working electrode. The system was tested on 10 selected wine samples, and the results were compared with total reactive antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), and chemiluminescent determination of total antioxidant capacity (CL-TAC) methods using Trolox and catechin as standards. Microdialysis online-coupled with amperometric detection gives similar results to the widely used cyclic voltammetry methodology and closely correlates with ORAC and TRAP. The problem of electrode fouling is overcome by the introduction of an electrochemical cleaning step (1-2 min at the potential of 0 V vs Ag/AgCl). Such a procedure is sufficient to fully regenerate the electrode response for both red and white wine samples as well as catechin/Trolox standards. The appropriate size of microdialysis probes enables easy automation of the electrochemical TAC/TCP measurement using 96-well microtitration plates.

Black and green tea - luminol false-negative bloodstains detection.

The antioxidant properties of black and green teas are well known. It is also possible to determine their antioxidant capacity by using a chemiluminscent method. This method is based on the measurement of the delay in the emission of light from the luminol reaction in the presence of the antioxidant. Bloodstains which are invisible to the naked eye can also be detected by luminol. Three common methods (detection using the Grodsky or Weber formulations and by Bluestar® Forensic latent bloodstain reagent) are based on the luminol chemiluminescence reaction. The bloodstains can be masked by drinks and/or foods containing antioxidants. The aim of this work was to compare the ability of black and green teas containing antioxidants to cause false negative results during chemiluminescent bloodstain detection.

Sodium azide as a specific quencher of singlet oxygen during chemiluminescent detection by luminol and Cypridina luciferin analogues.

Reactive oxygen species (ROS) are presently thought to play important role in an increasing number of the physiological and pathological processes in living organisms. Various chemiluminescent (CL) compounds have been studied in order to find suitable and specific probes for the detection of particular ROS species. The CL of luminol is known to be non-specific and can be induced by various oxidants. Two Cypridina luciferin analogues, CLA and MCLA, have been used for the detection of ROS in vivo. CLAs are thought to emit light only when reacting with superoxide and singlet oxygen. It is possible to distinguish the particular ROS by using a specific quencher or scavenger, e.g. superoxide dismutase (SOD) or sodium azide (NaN(3)). The CL reactions of luminol (3-aminophthalhydrazide), CLA [2-methyl-6-phenyl-3,7-dihydroimidazo(1,2α) pyrazin-3-one] and MCLA [2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo(1,2α) pyrazin-3-one] were studied in three hydrogen peroxide decomposition systems (H(2)O(2)-HRP; H(2)O(2)-CuSO(4); and H(2)O(2)-NaOCl). The measurements were carried out in phosphate buffer, pH 7.4, at 25°C, using a luminometer (Fluoroskan Ascent FL and Sirius C). NaN(3) was used as the specific quencher of singlet oxygen. The results demonstrate that the proclaimed specificity of the CL of Cypridina luciferin analogues towards singlet oxygen has to be discussed.

The photodynamic effect: the comparison of chemiexcitation by luminol and phthalhydrazide.

The presence of light, oxygen and photosensitizer (organic dye) is required for the photodynamic effect. Light and photosensitizer are harmless by themselves, but when combined with oxygen, reactive oxygen species (ROS) can be produced. This photodynamic effect is used in photodynamic therapy (PDT); the production of ROS as lethal cytotoxic agents can inactivate tumor cells. However, during PDT, there are many difficulties, so it is not possible to excite the photosensitizer using a laser, a source of light at the wavelengths specific to the photosensitizer (in visible region of the spectrum). Chemiluminescence is the light emission as a result of a chemical reaction. It is possible to use a chemiluminescent mixture to excite the photosensitizer even if the light emission does not conform to the absorption maximum of the photosensitizer. Luciferin and luminol have been used as chemiluminescent compounds (energizers) for the excitation of the photosensitizers. The aim of this work was to compare the chemiexcitation of some selected photosensitizers (e.g. fluorescein, eosin, methylene blue, hypericin and phthalocyanines) by chemiluminescent mixtures containing luminol (high chemiluminescent quantum yield) or phthalhydrazide (low chemiluminescent quantum yield) on some Gram-positive (Enterococcus faecalis, Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa, E. coli) bacteria and some cell lines (NIH3T3 and MCF7). The efficiency of the chemiexcitation was dependent on the kind of the photosensitizer and on the type of the bacterial strain or cell line and was independent of the energizers.