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At least 343 records · Page 19Linked to original sources
PubMed2005

Chemiluminescence determination of amikacin based on the inhibition of the luminol reaction catalyzed by copper.

A simple and sensitive method has been proposed for the amikacin sulphate determination. It is based on the inhibition of the chemiluminescence (CL) emission generated from the oxidation of luminol in alkaline medium by H2O2 catalyzed by Cu(II), due to the interaction caused by amikacin, which forms a robust complex with the catalyst. The optimization of the experimental and instrumental variables affecting this CL inhibition effect has been carried out using statistical models, based on the application of two-level full factorial and Box-Behnken designs. The performance characteristics of the proposed method have been established, showing that the method is efficient to determine amikacin sulphate in the linear range of 9.89-20 mg/L with a detection limit of 2.97 mg/L. It has been successfully applied to the amikacin sulphate determination in pharmaceutical formulations.

Amikacin↗
PubMed2006

Flow injection determination of adenine at trace level based on luminol-K2Cr2O7 chemiluminescence in a micellar medium.

A novel flow injection chemiluminescence (CL) analysis method for the determination of adenine in the presence of sodium dodecylbenzene sulfonate (SDBS) surfactant micelles is described. This method is based on the luminescent properties of the luminol-K2Cr2O7-adenine in alkaline medium sensitized by SDBS. The optimized experimental conditions were evaluated and the possible mechanism was discussed by examining CL emission spectrum and the effect of various free radical scavengers on CL emission intensity. The CL increase is linearly related to the concentration of adenine in the range of 2.92x10(-6) to 4.38x10(-10) mol l-1 with a detection limit of 2.46x10(-10) mol l-1 (S/N=3). The relative standard deviation for 2.92x10(-7) mol l-1 samples was 1.67% (n=12). The proposed method has been applied to the determination of adenine in human serum.

Adenine↗
PubMed2006

Rapid determination of clindamycin in medicine with myoglobin-luminol chemiluminescence system.

A sensitive chemiluminescence method, based on the inhibitory effect of clindamycin on the chemiluminescence reaction between luminol and myoglobin in a flow-injection system, is proposed for the determination of clindamycin. The decrement of chemiluminescence intensity is linear with the logarithm of clindamycin concentration over the range from 0.1 to 70.0 ng ml-1 (r2=0.9995), with the detection limit of 0.03 ng ml-1 (3sigma). At a flow rate of 2.0 ml min-1, a complete analytical process could be performed within 0.5 min, including sampling and washing, with a relative standard deviation of less than 3.0% (n=5). The proposed procedure was applied successfully to the determination of clindamycin in capsules without any pretreatment process.

Anti-Bacterial Agents↗
PubMed2004

Flow injection determination of p-aminophenol at trace level using inhibited luminol-dimethylsulfoxide-NaOH-EDTA chemiluminescence.

A novel flow injection procedure was developed for the determination of p-aminophenol (PAP) based on the inhibition by PAP of the chemiluminescence from luminol-dimethylsulfoxide (DMSO)-NaOH-EDTA system. The method has merits of higher sensitivity, wider linear range, simpler procedure, and a more rapid analyzing speed. It is applicable for the determination of PAP in the range of 2.5 x 10(-10)-5.0 x 10(-8) g mL(-1) with a detection limit of 1.9 x 10(-10) g mL(-1). The relative standard deviation (RSD) for 5.0 x 10(-9) g mL(-1) PAP is 0.78% (n=15). The method has been successfully used to determine PAP in industrial wastewaters and environmental waters. Additionally, the inhibition mechanism was also discussed briefly.

Aminophenols↗
PubMed1997

Correlation between arachidonic acid oxygenation and luminol-induced chemiluminescence in neutrophils: inhibition by diethyldithiocarbamate.

Neutrophils from allergic subjects were hypersensitive to stimulation by low calcium ionophore concentration (0.15 microM), resulting in an increased formation of leukotriene B4 (LTB4), 5S-hydroxy-6,8,11,14-(E,Z,Z,Z)-eicosatetraenoic acid (5-HETE), and other arachidonic acid metabolites through the 5-lipoxygenase pathway. In parallel, luminol-dependent chemiluminescence was also higher in neutrophils from allergic patients at the basal state and after stimulation by calcium ionophore, revealing an enhancement of radical oxygen species and peroxide production. The activity of glutathione peroxidase, the main enzyme responsible for hydroperoxide reduction, was lowered in these cells. Diethyl-dithiocarbamate (DTC) induced a concentration-dependent decrease in chemiluminescence and arachidonic acid metabolism after neutrophil stimulation. These data show that the elevation of arachidonic acid metabolism in neutrophils from allergic patients is strongly correlated with oxidative status. This elevation may be the consequence of an increased cellular hydroperoxide known to activate 5-lipoxygenase (5-LOX) activity and/or an increased arachidonic acid availability, due either to phospholipase A2 (PLA2) activation or inhibition of arachidonate reesterification into phospholipids. Lowering this oxidative status was associated with a concomitant decrease of this metabolism. Our results suggest that the effect of DTC may be the consequence of an inhibition of peroxyl radical and cellular lipid hydroperoxide production. Thus, DTC may modulate arachidonic acid metabolism in neutrophils by modulating the cellular hydroperoxide level.

Adult↗
PubMed1997

Analysis and assessment of the capacity of neutrophils to produce reactive oxygen species in a 96-well microplate format using lucigenin- and luminol-dependent chemiluminescence.

The chemiluminescence (CL) assay has been used to measure the reactive oxygen species (ROS)-generating capacity of phagocytes. To achieve more optimal measurement conditions for a multi-channel microplate photon-counting CL analyzer with the cooled charge-coupled device (CCD) camera which offers enhanced sensitivity, we investigated factors affecting the variability in lucigenin-dependent CL (LgCL) measurement of human neutrophils stimulated with either opsonized zymosan (OZ) or phorbol myristate acetate (PMA). We obtained sensitive LgCL responses with good reproducibility and rapid data-acquisition using 50 microl neutrophils (3 X 10(6) cells/ml) and 50 microl of 0.5 mM lucigenin per well, in addition to either 100 microl of OZ (5 mg/ml) when zymosan was opsonized with 10-20% serum or 100 microl of PMA solution (1 X 10(-6) M) with automatic regular intervals of mixing and detection during the continuous measurement at 37 degrees C. Furthermore, we studied the contribution of various ROS to LgCL and luminol-dependent CL (LmCL) using modulators of ROS metabolism including superoxide dismutase (SOD), catalase, deferoxamine and sodium azide (NaN3). LgCL was inhibited by SOD but not by the other agents, whereas LmCL was inhibited by NaN3 and deferoxamine. Thus, it was demonstrated that LgCL detects the superoxide anion with high selectivity whereas the LmCL assay measures myeloperoxidase (MPO)-mediated formation of hypochlorous acid. Such microplate-based multiple measurements facilitate the accurate assessment of phagocytic function.

Acridines↗
PubMed1996

A new reliable chemiluminescence immunoassay (CLIA) for prostaglandin E2 using enhanced luminol as substrate.

A sensitive and reliable chemiluminescence immunoassay suitable for the quantitative determination of prostaglandin E2 (PGE2) has been developed using 96 well microtiter plates (MTP). The assay is based on a competitive reaction between a highly specific monoclonal anti-PGE2 antibody (mouse), free antigen and solid phase bound antigen. The MTP was first coated with a bovine serum albumin (BSA)-PGE2 conjugate. Then, after preincubating, the anti-PGE2 antibody (Ab) and the analyte were added. The remaining amount of free antibody was captured by the solid phase bound BSA-PGE2 conjugate. The monoclonal antibody captured on the MTP was determined using biotinylated anti-mouse-Ab and a complex of avidin and biotin-labelled horseradish peroxidase (HRP). Substrate for HRP was the cyclic diacyl hydrazide compound luminol, enhanced by p-iodophenol. Photons emitted during the reaction were measured using a photomultiplier tube. The assay has been validated with assay buffer and human plasma over a concentration range of 10-50,000 pg/ml. The lower limit of quantification is 100 pg/ml (2 pg/well) and 150 pg/ml (3 pg/well) for buffer and plasma, respectively. The intra-day coefficients of variation (CV) for the range of 100-50,000 pg/ml are 3.2-8.9% (buffer) and 4.2-17.7% (plasma) and inter-day CV are 2.9-19.8% (buffer) and 3.6-21.2% (plasma). The method can be used for quantification of PGE2 in biological fluids like plasma and suction blister fluid.

Animals↗
PubMed1985

Variant specific opsonization of Trypanosoma evansi measured by luminol-dependent chemiluminescence.

Using luminol-dependent chemiluminescence (LCL), the specificity of antibodies to variable antigen type (VAT)-populations of Trypanosoma evansi was studied in four infected ponies. Trypanosomes of each wave of parasitemia were isolated and multiplied in irradiated mice. Their opsonization by serum collected during the infection was investigated with LCL and results for isolated VAT-populations are shown in the paper. Antibodies specific to each VAT-population were first found three days after the maximum of a parasitemic wave. There was no cross reactivity between different VAT-populations. LCL proved to be a rapid and automatic method for the demonstration of antibodies with specificity to variable antigen types of trypanosomes.

Animals↗
PubMed1983

Carbonate anions; effects on the oxidation of luminol, oxidative hemolysis, gamma-irradiation and the reaction of activated oxygen species with enzymes containing various active centres.

Presence of carbonate anions increases the oxidation of luminol in different chemical systems. Lysis of human erythrocytes due to the action of dihydroxyfumaric acid or of perborate is also stimulated by carbonate ions. These anions also change considerably the loss of activity of different enzymes treated with superoxide, hydroxyl or formate radicals and can increase or decrease the effect as a function of the nature of the active centre of the enzyme. The relative effects of superoxide, hydroxyl, formate and carbonate radicals for the inactivation of various enzymes (superoxide dismutases, catalase, ribonuclease, glucose oxidase and glutathione peroxidase) have been examined. Three systems were used: gamma-irradiation under different conditions, photoproduction of radicals and sonication. Inactivation of the enzymes is a function not only of the radical used but also of the nature of the active site. Thus glutathione peroxidase is remarkably resistant to hydroxyl radicals while the superoxide dismutases are rapidly inactivated by carbonate radicals. All of the results combine to show that the presence or absence of carbonate anions must be considered in all studies of oxygen containing free radicals whether chemical, biochemical or biological or high energy irradiation.

Animals↗
PubMed1997

Real-time measurement of nitric oxide production in rat brain by the combination of luminol-H2O2 chemiluminescence and microdialysis.

A chemiluminescence method of detecting nitric oxide (NO) in combination with a microdialysis technique was employed for the real-time measurement of NO production in living rat brain. This method based on the luminol-H2O2 system has a detection limit of 1 nM, and is the most sensitive method currently available for measuring NO. We applied this new technique to rat cerebellum to record the increase of chemiluminescence intensity arising from NO production after the injection of N-methyl-D-aspartate or kainic acid around the microdialysis probe. This highly sensitive method should be useful for the direct clarification of the functions of NO in the central nervous system.

Animals↗
PubMed2000

Attenuation of luminol-amplified chemiluminescent intensity and lipid peroxidation in the livers of quercetin-fed mice.

We assessed the antioxidative effects of quercetin-feeding on ddY strain male mice by measuring luminol-amplified chemiluminescence that was enhanced by a hydrophilic free radical initiator 2, 2'-azobis(2-amidinopropane) dihydrochloride. Quercetin suppressed chemiluminescent intensity in a dose-dependent manner in vitro with a half-inhibition concentration (IC(50)) of 3x10(-8) M, which was lower than the value of synthetic antioxidant 2, 6-di-tert-butyl-p-cresol (IC(50): 7x10(-7) M). Lysosomal (12000xg pellet) and supernatant fractions obtained from the livers of mice fed a diet containing 0.2% quercetin for 7 days showed more inhibition of chemiluminescent intensity than those from non-treated mice. Quercetin feeding also resulted in the inhibition of lipid peroxidation as determined by a thiobarbituric acid reactive substance test which detected suppression of the release of lysosomal enzymes induced by lipophilic free radical initiator 2, 2'-azobis(2,4-dimethylvaleronitrile). These results probably reflect radical quenching activity of quercetin, indicating that the measurement of chemiluminescence is a useful tool for the assessment of total antioxidant activity in biological systems.

Acetylglucosaminidase↗
PubMed1996

Luminol-and lucigenin-amplified chemiluminescence with rat liver microsomes. Kinetics and influence of ascorbic acid, glutathione, dimethylsulfoxide, N-t-butyl-a-phenyl-nitrone, copper-ions and a copper complex, catalase, superoxide dismutase, hexobarbital and aniline.

For the investigation of luminol (LM)-and lucigenin (LC)-amplified chemiluminescence (CL) in rat liver microsomes using both a liquid-scintillation counter (LKB/Wallac 1219 Rackbeta) and a Berthold luminometer (AutoLumat LB 953) optimal incubation mixtures and conditions and basic kinetics have been established. Whereas calibration curves for both LM- and LC-CL are performed with hydrogenperoxide (LC quantum yield is 6.25 fold higher as that of LM), distinct differences were revealed with microsomes, indicating that different reactive oxygen species (ROS) are determined: Both LM- and LC-CL follow the kinetics of enzymatic reactions in terms of dependence on protein and NADPH or NADH concentration, time course, temperature etc., but with differences. LM-CL does not work without addition of Fe2+, whereas LC-CL does. Both copper ions and copper bound in a complex abolish CL, LC-CL being much more sensitive. Isolated cytochrome P-450 (P450) and NADPH P450 reductase from liver of pheno-barbital treated rats alone proved to be inactive in LM-and LC-CL production, whereas te combination 1:1 without and with addition of lipid was highly active in both LM-and LC-CL. Ascorbic acid and glutathione as scavengers diminish both LM- and LC-CL in concentrations higher then 10(5). Dimethyl-sulfoxide (DMSO) was ineffective in LM-CL up to concentrations of 0.2 M, the very high concentration of 2 M diminished LM-CL only to 1/3. LC-CL was diminished starting at concentrations of 100 mM and at 2 M only 10% of maximum LC-CL was observed. The trap substance N-t-butyl-a-phenylnitrone (BNP) also diminished LC-CL more effectively than LM-CL. Clearcut differences were revealed by the addition of catalase and superoxide dismutase: both enzymes diminished LM-CL only, without any influence on LC-CL. Hexobarbital, a potent uncoupler of P450, enhances LM-CL fivefold, whereas LC-CL is barely influenced. Aniline (without uncoupling capability) decreased both LM-and LC-CL increasingly with increasing concentrations. Therefore the conclusion is drawn that LM-CL measures in liver microsomes predominantly superoxide anion radicals, whereas LC-CL is mainly a measure for microsomal hydroxyl radical formation or of reactive organic radicals. With microsomes of phenobarbital and beta-naphthoflavone treated rats CL was much higher but in principle the same kinetic characteristics could be shown. All results on microsomes were obtained uniformly with the liquid scintillation counter and the Berthold luminometer, the letter being much more effective and more sensitive.

Acridines↗
PubMed1998

Luminol and lucigenin amplified chemiluminescence and lipidperoxidation with brain microsomes from rats during ontogenetic development.

The chemiluminescence (CL) amplifiers luminol (LM) and lucigenin (LC) react with different reactive oxygen species (ROS) in dependence on the ROS generating system used. With liver microsomes LMCL indicates predominantly superoxide anion radicals, whereas LCCL is mainly a measure for hydroxyl radical formation or of reactive organic radicals. With brain microsomes only LCCL, but not LMCL could be measured. For both brain microsomes from newborn (both sexes) and 60 day-old (male) rats LCCL is dependent on protein and NADPH concentration, activity in newborns being only 15% compared with young adult rats. As compared with liver microsomes 10-fold higher protein concentrations are needed to obtain comparable LCCL, whereas the NADPH demand is the same as with liver. A distinct ontogenetic development was demonstrated: low activities in the fetus, in newborn and 10-day-old rat are followed by higher activities with increasing age, after a maximum at an age of 60 days a decline was observed. Microsomal lipidperoxidation (LPO) was measured as formation of thiobarbituric acid reactive substances (TBARS) and was also dependent on protein and NADPH concentration. Unexpectedly, LPO with brain microsomes from newborn rats did not show any developmental variation.

Acridines↗
PubMed2003

Evaluation of the antioxidant effect of melatonin by flow injection analysis-luminol chemiluminescence.

The antioxidant effect of melatonin was evaluated by using a flow injection analysis chemiluminescence (FIA-CL) method to measure alone its direct antioxidant behaviour in aqueous solutions. Inhibition by melatonin, DL-tryptophan or serotonin (5-hydroxytryptamine, 5-HT) of the steady CL emissions of various oxidants with luminol was measured. LogIC(50) values of melatonin were calculated: -2.6+/-0.1M for superoxide (O(2) radical(-)), -3.0+/-0.02 M, for hydrogen peroxide (H(2)O(2)), -4.7+/-0.04 M, for hypochlorous anion ((-)OCl), -2.8+/-0.1M, for hydoxyl (radical OH) and -2.0+/-0.03 M, for peroxynitrite (ONOO(-)). Thus melatonin exerted a limited direct antioxidant activity towards reactive oxygen species (ROS) in aqueous solutions, broadly comparable to that of tryptophan, but less than that of serotonin and of major antioxidants such as ascorbic acid. This suggests that where the benefits of melatonin are correctly attributed to antioxidant behaviour, this is mediated largely through antioxidant enzymes. The pharmacological implications are discussed.

Antioxidants↗
PubMedDate not supplied

A transition metal enhanced luminol chemiluminescence in the presence of a chelator.

We have investigated the chemiluminescence signal of luminol and hydrogen peroxide in the presence of a transition metal (Co(II), Cu(I), Fe(II), Fe(III)) and of a chelator (EDTA, citric acid) in pH 8.5, 9 and 10 borate buffer solutions. We observed that the chemiluminescence intensities of these systems reached a plateau, where they remained stable for a period of 2-30 s. We also observed linearity between the intensity of chemiluminescence and the hydrogen peroxide concentration. The combination of Co(II) and EDTA at pH 9 was found to give the optimum signal with reference to time stability, intensity and reproducibility. Thus, compared to previous chemiluminescence applications, the present results permit us to propose a simple, enzyme-free and time-independent technique for the detection and quantification of hydrogen peroxide.

Buffers↗
PubMed2001

Immunomodulation by exogenous surfactant: effect on TNF-alpha secretion and luminol-enhanced chemiluminescence activity by murine macrophages stimulated with group B streptococci.

Group B streptococci (GBS) are important pathogens in neonatal sepsis and pneumonia. GBS stimulate alveolar macrophages to produce inflammatory cytokines and free oxygen radicals, which can damage the lungs. In several studies, use of exogenous surfactant in term babies has improved outcome related to sepsis and respiratory failure. The role(s) of exogenous surfactant in modulating the inflammatory response produced by this microbe was examined. Tumor necrosis factor alpha (TNF-alpha) production and luminol-enhanced chemiluminescence (LCL), a measure of respiratory burst, were investigated. For measuring TNF-alpha release, RAW 264.7 murine macrophages were pre-incubated with bovine surfactant and stimulated with either lipopolysaccharide, live or heat-killed GBS type Ia. LCL was measured after macrophages were pre-incubated with or without surfactant overnight, then stimulated with GBS or phorbol myristate acetate. Lipopolysaccharide and GBS stimulated TNF-alpha secretion from macrophages that was suppressed by exogenous surfactant in a dose-dependent fashion. GBS and phorbol myristate acetate also increased LCL from macrophages, which was significantly suppressed by pre-incubation of macrophages with exogenous surfactant. We conclude that GBS type Ia stimulates TNF-alpha release and LCL from RAW 264.7 cells and that these responses are suppressed by surfactant. Suppression of inflammatory mediators by exogenous surfactant might improve respiratory disease associated with GBS.

Animals↗
PubMed2001

Luminol-dependent chemiluminescence analysis of chitooligosaccharide-induced rapid production of hydrogen peroxide by intact wheat seedlings.

The feasibility of a luminol-dependent chemiluminescence analysis of hydrogen peroxide production by intact wheat seedlings using a KhL-003 chemiluminometer was determined. It was shown that the minimal H2O2 concentration that can be detected in a 0.5-ml sample with this instrument is 0.125 microM. Analysis of biological activity of a mixture of chitooligosaccharides with molecular masses from 5 to 10 kD and acetylation degree of 65% demonstrated that, at a concentration of 1 microg/ml, they induce rapid overproduction of H2O2 in roots of 3-day-old wheat seedlings.

Chitin↗
PubMed2004

p-Iodophenol-enhanced luminol chemiluminescent assay applied to discrimination between acute lymphoblastic and minimally differentiated acute myeloid (FAB-M0) or acute megakaryoblastic (FAB-M7) leukemias.

INTRODUCTION: In this report, we propose the application of the p-iodophenol-enhanced luminol chemiluminescent technique to the determination of peroxidase (myeloperoxidase and/or platelet peroxidase) activity in blasts of minimally differentiated acute myeloblastic leukemia (AML-M0) and acute megakaryoblastic leukemia (AML-M7). METHODS: The frozen blast cells from 29 patients were thawed and submitted to the optimized protocol. RESULTS: All cases of AML-M7 and AML-M I exhibited integrated light emission greater than 73(10(2) mV x s), which was the arbitrary cutoff point set for the discrimination between AML and acute lymphoblastic leukemia (ALL) (mean + 3 x s.d. of ALL samples, n = 10). In addition, five out of seven cases of AML-MO showed results above the cutoff point. CONCLUSION: This highly sensitive enhanced chemiluminescent technique may be applied to discriminate between ALL and AML-M7 or AML-MI cases, and most AML-M0 cases. It is very simple, cheap and easy to perform compared to other procedures used to measure MPO activity in AML-leukemias including AML-M7 and AML-M0.

Bone Marrow Cells↗