Synergetic Dual-Site Atomic Catalysts for Sensitive Chemiluminescent Immunochromatographic Test Strips.

In view of the outstanding catalytic efficiency, single-atom catalysts (SACs) have shown great promise for the construction of sensitive chemiluminescent (CL) platforms. However, the low loading amount of active sites dramatically obstructs the improved catalytic activity of these metal SACs. Benefiting from the exceedingly unique catalytic properties of the metal-metal bonds, atomic clusters may give rise to enhancing the catalytic properties of SACs based on the synergistic effects of dual atomic-scale sites. Inspired by this, atomic Co3N clusters-assisted Co SACs (Co3N@Co SACs) were synthesized through a facile doping method. Through X-ray absorption spectroscopy, the active metal sites in the synergetic dual-site atomic catalysts of Co3N@Co SACs were confirmed to be Co-O4 and Co3-N moieties. Co3N@Co SACs served as a superior co-reactant to remarkably enhance the luminol CL signal by 2155.0 times, which was prominently superior to the boosting effect of the pure Co SACs (98.4 times). The synergetic dual-site atomic catalysts contributed to accelerating the decomposition of H2O2 into singlet oxygen as well as superoxide radical anions to display superb catalytic performances. For a concept employment, Co3N@Co SACs were attempted to utilize as CL probes for establishing a sensitive immunochromatographic assay to quantitate pesticide residues, in which imidacloprid was adopted as the model analyte. The quantitative range of imidacloprid was 0.05-10 ng mL-1 with a detection limit of 1.7 pg mL-1 (3σ). Furthermore, the satisfactory recovery values in mock herbal medicine samples demonstrated the effectiveness of the proposed Co3N@Co SAC-based CL platform. In the proof-of-concept work, synergetic dual-site atomic catalysts show great perspectives on trace analysis and luminescent biosensing.

A Smart Advanced Chemiluminescence-Sensing Platform for Determination and Imaging of the Tissue Distribution of Natural Antioxidants.

Antioxidants have gained marked attention owing to their ability to prevent the oxidation of biological components and to protect the body from reactive oxygen species, thereby maintaining human health. Thus, antioxidant-rich dietary supplements and natural foods can be effective against oxidative stress and can even act as chemopreventive agents. Therefore, a simple and rapid assay for evaluation of antioxidant capacity and assessment of their distribution profile in natural sources is vital. Herein, we report a rapid, innovative chemiluminescence (CL) platform for evaluation and visualization of antioxidant capacity. We found that intense and long-lasting CL was formed upon the redox reaction of quinones, e.g., menadione, with antioxidants, e.g., l-ascorbic acid, in the presence of luminol. The produced CL intensities were proportional to the antioxidants' concentrations with a detection limit of 0.18 µM for the model antioxidant, l-ascorbic acid. As the formed CL was long-lasting, it could be easily captured and detected with a charge-coupled device (CCD) camera. To evaluate the quantification ability of the CCD camera, we developed a smart and fast microplate-based assay based on photographing the generated CL with a cooled CCD camera. The photographed CL intensities were linearly proportional with the antioxidant concentrations, and then the method was applied for photographing multiple food sample extracts. Ultimately, we utilized our method for the distribution profiling of antioxidant capacity in food cut sections. Samples were dipped in luminol and then in quinone, followed by CCD camera photography, without the need for any pulverization/extraction procedure, giving precise antioxidant distribution information.

Dual-modality probe based on black phosphorous and NiFe2O4 NTs for electrochemiluminescence and photothermal detection of ovarian cancer marker.

An electrochemiluminescence and photothermal immunosensor based on a dual-modality integrated probe was proposed for sensitive and reliable detection of lipolysis stimulated lipoprotein receptor (LSR), a new biomarker of ovarian cancer. Black phosphorous quantum dots (BPQDs) possess fascinating electrochemical property and unique photothermal effect, which could not only enhance ECL signal of N-(4-aminobutyl)-N-ethylisoluminol (ABEI) through accelerating dissolved O2 evolution but also realize temperature signal output by converting laser energy into heat. Furthermore, NiFe2O4 nanotubes (NiFe2O4 NTs) have large specific surface area and favorable adsorption ability, which could increase the immobilized amount of ABEI and BPQDs, further strengthening ECL and temperature signal. As a result, a dual-mode immunosensor was constructed and realized ECL and temperature dual signal to detect LSR, making the results more reliable. This work provided a new thought for the development of sensitive and accurate sensors and was expected to employ for determination of other biomarkers.

On the use of acridinium indicators for the chemiluminescent determination of the total antioxidant capacity of dietary supplements.

Acridinium salts, due to their chemiluminogenic properties, have found several applications in biomedical analysis as labels and indicators, where the assessment of emission intensity is used for the end-point detection. This work presents the use of chemiluminescent indicators in the form of selected acridinium esters in order to determine the antioxidant properties of exemplary formulations, namely quercetin, vitamin C and the dietary supplement, Apiextract. The principle of measurements is based on a change in the kinetics of emission decay derived from the acridinium cations in alkaline solutions of hydrogen peroxide in the presence of an antioxidant (the analyte). The proposed system makes a beneficial alternative to related methods, which mostly rely on the assessment of emission efficiency and use the luminometric standard luminol - due to superior parameters of acridinium chemiluminescence, among others - high temporary emission efficiency. The features of the proposed method are manifested by a shorter time period of analysis and lower background signals associated with the environmental influences, as compared to typical approaches. The chromatographic (RP-HPLC) analyses of the substrates and products generated during chemiluminogenic oxidation of acridinium cations under assay conditions are also presented.

Simple, Fast, and Sensitive detection of artemisinin in human serum and Artemisia annua using microsensor array coupled with electrochemiluminescent imaging technique.

Artemisinin is an important frontline antimalarial. Fast, accurate detection of artemisinin in human serum is of importance in monitoring its clinical pharmaceutical effect. In this work, a strategy using microsensor array coupled with electrochemiluminescence (ECL) imaging technique was developed for detection of artemisinin. The microsensor array was constructed by integrating a patterned indium tin oxide glass plate with two perforated hydrophobic paper covers. By introducing the reactant of p-aminophenylboronic acid, luminol and artemisinin into the microsensor array, artemisinin would oxidize p-aminophenylboronic acid into p-aminophenol, a product which can efficiently inhibit the ECL of luminol. ECL signals decrease linearly with the increase of artemisinin. Based on the decreased ECL signal, artemisinin can be accurately detected. A good linearity (r = 0.994) was observed for artemisinin detection. The detection sensitivity is 0.48 µM for artemisinin. The detection selectivity and stability were also investigated. Results show that the present method shows a good selectivity and stability towards artemisinin detection. To evaluate the applicability of the present strategy for detecting artemisinin in real samples, the artemisinin content in human serum and Artemisia annua samples were analyzed. Results demonstrated that the present strategy shows excellent selectivity with high sensitivity towards artemisinin detection in real samples.

A fast and simple FIA-chemiluminescence method for the evaluation of Roselle flowers as scavenger of the free radicals generated by UV irradiated antibiotics.

This work proposes a new method for the in vitro evaluation of the effect of UV irradiation on the production of free radicals and other reactive species during the photodecomposition of drugs. The method was based on the UV irradiation of antibiotics molecules to generate excited states that undergo to homolytic bond cleavages. These reactive species can be detected by their ability to oxidize the luminol, producing the electronically excited aminophtalate, which decays to the ground state releasing electromagnetic radiation in the visible zone of the spectrum. This method was applied to penicillin G, nafcillin, azlocillin and neomycin dissolved in water. It was found that the intensity of the luminol chemiluminescence emission (CL) was proportional to the concentration and dependent on the molecular structure of these drugs. Under the optimized conditions, it was found that penicillin and azlocillin were the most susceptible to photodegradation, while neomycin sulfate was the less affected by the UV light. It was observed that the addition to the antibiotics dissolutions of a hydro-alcoholic extract of petals of calyxes of Roselle reduced the CL intensity, indicating that the extract was able to scavenge the free radicals in the irradiated drugs. This result suggest that its addition to the antibiotics can help in the protection against the radicals formed during the exposition to solar light of patients treated with topic similar antibiotics.

A novel luminol-based enhanced chemiluminescence antioxidant capacity microplate assay for use in different biological matrices.

INTRODUCTION: Reactive oxygen species (ROS) are normal metabolic products of living cells. However, a decrease of the defense mechanisms against the effects of ROS or increased ROS production maybe one important causative factor of cellular damage. A non-enzymatic scavenger system is considered to be responsible for the maintenance of total antioxidant capacity (TAC) as a protection against oxidative injuries that exist in all higher plants and in mammals as well. METHODS: In our work, we optimized and validated a luminol-peroxidase-4-iodophenol-H2O2 enhanced chemiluminescence-based (ECL) TAC measurement technique. BSA was applied in the reagent to prevent peroxidase from auto-oxidation. The ECL method was suitable for plant extracts and for human blood serum as well. Our TAC technique was adapted to microplates and compared to ORAC assay using plant extracts. RESULTS AND DISCUSSION: The ECL method is fast (10min) with an interassay precision of <10% as CV. TAC values of ethanolic extracts of 10 plant species did correlate (ECL vs ORAC assay data: r=0.84, 95% confidence interval, CI=0.78-0.89, P<0.001) but with systematic bias. Analysis of serum samples obtained from septic and control patients showed significantly higher TAC values in the patient group compared to those of controls (p<0.01). Moreover, we could discriminate between surviving and non-surviving patients, based on their TAC values (p<0.01). Pearson's statistics showed the strongest positive correlation with serum uric acid (r=0.73). Besides the routine laboratory parameters, our novel TAC method might give complementary information on the severity of systemic inflammation.

An ultra-facile and label-free immunoassay strategy for detection of copper (II) utilizing chemiluminescence self-enhancement of Cu (II)-ethylenediaminetetraacetate chelate.

The establishment of facile, rapid, sensitive and cost-effective protocols for the detection of heavy metals is of great significance for human health and environmental monitoring. Hereby, an ultra-facile and label-free immunoassay strategy was designed for detecting heavy metal ion by using Cu (II) as the model analyte. Cu (II) reacted previously with ethylenediaminetetraacetate (EDTA) was captured by immobilized monoclonal antibody for Cu (II)-EDTA chelate. Then Cu (II) was detected based on the self-enhancing effect of Cu (II)-EDTA chelate to luminol-H2O2 chemiluminescence reaction. The CL intensity is linear relative with Cu (II) concentration in a very wide range of 1.0-1000ng/mL, with a detection limit of 0.33ng/mL (S/N=3). Since the specificity of this proposed strategy relied on both the specificity of monoclonal antibody and the specificity of luminol-H2O2 system, it could avoid interference from most common ions. The proposed method was used successfully to detect Cu (II) in traditional Chinese medicine and environmental water samples with acceptable recovery values of 82-113%. This proof-of-principle work demonstrated the feasibility of the label-free immunoassay for heavy metal ions, and opened a new avenue for rapid screening and field assay for drug safety, environmental monitoring and clinical diagnosis.

Screening of peroxynitrite scavengers in Flos Lonicerae by using two new methods, an HPLC-DAD-CL technique and a peroxynitrite spiking test followed by HPLC-DAD analysis.

INTRODUCTION: Peroxynitrite is involved in the pathogenesis of a number of significant diseases. Peroxynitrite scavengers thus have potential application in understanding and treating these diseases. It is, therefore, important to establish screening methods able to rapidly identify peroxynitrite scavengers from herbal plants. OBJECTIVE: To develop effective and easily operable screening methods for identifying peroxynitrite scavengers in complex matrices, including Chinese herbal medicines. METHODS: Two simple and efficient screening methods have been developed for the identification of natural peroxynitrite scavengers in Flos Lonicerae Japonicae (FLJ). Method I used HPLC-DAD-(luminol-peroxynitrite)-CL techniques combined with Q-TOF MS/MS analysis, while Method II used pre-column reaction of the sample with peroxynitrite, followed by HPLC separation and Q-TOF MS/MS analysis. RESULTS: Five peroxynitrite scavengers (neochlorogenic acid, chlorogenic acid, 3,4-O-dicaffeoyl quinic acid, 3,5-O-dicaffeoyl quinic acid and 4,5-O-dicaffeoyl quinic acid) were identified in FLJ using Method I. Besides the compounds identified using Method I, three additional peroxynitrite scavengers (rutin, isoquercitrin and luteoloside) were identified using Method II. CONCLUSION: The two new methods proved to be complementary and the use of these methods should allow rapid detection of peroxynitrite-scavenging natural products from FLJ and other complex matrices.

[Study of Antioxidant and Membranotropic Activities of Quinazoline Alkaloid Tryptanthrin Using Different Model Systems].

A comparative study of antioxidant (radical-interceptor) properties of tryptanthrin (quinazoline alkaloid shows a high anti-inflammatory activity and it is found in many types of different families of higher plants and microorganisms, including the human microbiome) in the systems of 2,2'-azo-bis(2-methylpropionamidin)dihydrochloride-luminol and hemoglobin-hydrogen peroxide-luminol has been conducted and the influence on the permeability of planar bilayer lipid membranes is evaluated. Trolox was used as a reference antioxidant, and ascorbic acid and dihydroquercetin were taken as standards. Tryptanthrin exhibits very weak antioxidant activity, being markedly inferior to the reference standard and antioxidants while testing antioxidant activity in both studied systems. By the efficacy of antioxidative action the substrates in the systems studied can be arranged in the following order: dihydroquercetin > trolox > ascorbic acid > tryptanthrin. Antioxidant potential of tryptanthrin is approximately 1000 and 3000 times lower than that of trolox and bioflavonoid dihydroquercetine, respectively. Tryptanthrin causes no significant changes in the permeability of planar bilayer membranes in a dose range of 0.5 to. 10 µg/ml. Our data show that tryptanthrin displays no significant radical-interceptor and membranotropic activities. It can be assumed that the observed high anti-inflammatory activity of tryptanthrin is not related to the neutralizing effect against reactive oxygen species and the influence on the permeability of cell membranes. The anticipated mechanisms of biological activity of tryptanthrin are discussed.

Chemiluminescent aptasensor for chloramphenicol based on N-(4-aminobutyl)-N-ethylisoluminol-functionalized flower-like gold nanostructures and magnetic nanoparticles.

A novel chemiluminescent aptasensor for the highly sensitive detection of chloramphenicol (CAP) in milk was successfully developed using biotinylated CAP aptamer-functionalized magnetic nanoparticles (MNPs) as capture probes and thiolated hybridized complementary strand-modified N-(4-aminobutyl)-N-ethylisoluminol (ABEI)-functionalized flower-like gold nanostructures (AuNFs) as signal probes. P-iodophenol (PIP) was also added to form an ABEI-H2O2-PIP steady-state chemiluminescence (CL) system. Based on a competitive format, the CL intensity was negatively correlated with the concentration of CAP in the range of 0.01-0.20 ng/mL and the detection limit was 0.01 ng/mL in buffer and 1 ng/mL in milk. The proposed method was successfully applied to measure CAP in milk samples and compared to a commercial ELISA method. The high sensitivity of AuNFs, excellent selectivity and stability of aptamers, and good overall stability of the chemiluminescent bioassay with magnetic separation make them a promising approach for the detection of small molecular illegal additives. Additionally, the high sensitivity, easy operation, and good reproducibility exhibited by the stable chemiluminescent bioassay demonstrate its applicability for the trace detection of CAP in applications, such as animal husbandry.

A novel "dual-potential" electrochemiluminescence aptasensor array using CdS quantum dots and luminol-gold nanoparticles as labels for simultaneous detection of malachite green and chloramphenicol.

A novel type of "dual-potential" electrochemiluminescence (ECL) aptasensor array was fabricated on a homemade screen-printed carbon electrode (SPCE) for simultaneous detection of malachite green (MG) and chloramphenicol (CAP) in one single assay. The SPCE substrate consisted of a common Ag/AgCl reference electrode, carbon counter electrode and two carbon working electrodes (WE1 and WE2). In the system, CdS quantum dots (QDs) were modified on WE1 as cathode ECL emitters and luminol-gold nanoparticles (L-Au NPs) were modified on WE2 as anode ECL emitters. Then the MG aptamer complementary strand (MG cDNA) and CAP aptamer complementary strand (CAP cDNA) were attached on CdS QDs and L-Au NPs, respectively. The cDNA would hybridize with corresponding aptamer that was respectively tagged with cyanine dye (Cy5) (as quenchers of CdS QDs) and chlorogenic acid (CA) (as quenchers of l-Au NPs) using poly(ethylenimine) (PEI) as a bridging agent. PEI could lead to a large number of quenchers on the aptamer, which increased the quenching efficiency. Upon MG and CAP adding, the targets could induce strand release due to the highly affinity of analytes toward aptamers. Meanwhile, it could release the Cy5 and CA, which recovered cathode ECL of CdS QDs and anode ECL of L-Au NPs simultaneously. This "dual-potential" ECL strategy could be used to detect MG and CAP with the linear ranges of 0.1-100 nM and 0.2-150 nM, with detection limits of 0.03 nM and 0.07 nM (at 3sB), respectively. More importantly, this designed method was successfully applied to determine MG and CAP in real fish samples and held great potential in the food analysis.

Enzyme-linked immunosorbent assay by enhanced chemiluminescence detection for the standardization of estrogenic miroestrol in Pueraria candollei Graham ex Benth.

Miroestrol (ME) is a potent phytoestrogen from the P. candollei tuberous root. It has been approved for use in clinical trials due to its beneficial effect on disorders associated with estrogen deficiency. To ensure medical efficacy and safety, high performance analytical methods for ME analysis are required to standardize products from the P. candollei root. An enhanced chemiluminescence enzyme-linked immunosorbent assay (ECL-ELISA) was developed and validated using a polyclonal antibody against ME and a chemiluminescent system of luminol-H2 O2 -horseradish peroxidase-4-(1-imidazolyl) phenol. The ECL-ELISA system exhibited linearity over a concentration range of 0.31-10.00 ng mL(-1) , for which the relative standard variation (%RSD) was less than 10% for both intra- and interplate determinations. The ECL-ELISA is reliable for the determination of ME as reflected by the high recovery percentage (101.22-103.06%). As a comparative analysis, the ME content in each sample determined by ECL-ELISA was correlated with high coefficients of determination with colorimetric ELISA (R(2) = 0.998) and high performance liquid chromatography (HPLC) (R(2) = 0.998) methods. The ECL-ELISA method could be applied to all of the commercial products containing P. candollei root, when the products contain between 0.706 ± 0.046 and 13.123 ± 0.794 µg g(-1) dry wt. of ME. This method is useful as a high performance analytical method for the quantity control of ME in raw materials and end products at both the research and industrial levels.

Intensification of the electrochemiluminescence of luminol on hollow TiO2 nanoshell-modified indium tin oxide electrodes.

Hollow titania nanoshells (HTNSs), which were synthesized by a SiO2 sacrificial template method, were used to intensify the electrochemiluminescence (ECL) of luminol. The size, shell thickness and crystal phase, factors that are important in determining the efficiency, can be controlled by adjusting the template size, precursor concentration and calcination temperature, respectively. The structure of the HTNSs was characterized by transmission electron microscopy, scanning electron microscopy and X-ray diffraction spectroscopy. After structural optimization, the surface of indium tin oxide (ITO)-coated glass was modified with the HTNSs to act as a working electrode for a flow-injection analytical system. The heterostructure demonstrated an ECL emission intensity 150 times higher than that of the bare ITO. The research also revealed that the ECL of luminol on this modified electrode showed a very sensitive response to hydrogen peroxide with a detection limit of 4.6×10(-10)M. In addition to discussing the intensifying mechanism of luminol ECL by HTNSs, we demonstrate that can be successfully applied to evaluate the gross antioxidant activity of garlic.

Amplified electrochemiluminescence of luminol based on hybridization chain reaction and in situ generate co-reactant for highly sensitive immunoassay.

In this work, we described a simple and highly sensitive electrochemiluminescence (ECL) strategy for IgG detection. Firstly, L-cysteine functionalized reduced graphene oxide composite (L-cys-rGO) was decorated on the glassy carbon electrode (GCE) surface. Then anti-IgG was immobilized on the modified electrode surface through the interaction between the carboxylic groups of the L-cys-rGO and the amine groups in anti-IgG. And then biotinylated anti-IgG (bio-anti-IgG) was assembled onto the electrode surface based on the sandwich-type immunoreactions. By the conjunction of biotin and streptavidin (SA), SA was immobilized, which in turn, combined with the biotin labeled initiator strand (S1). In the presence of two single DNA strands of glucose oxidase labeled S2 (GOD-S2) and complementary strand (S3), S1 could trigger the hybridization chain reaction (HCR) among S1, GOD-S2 and S3. Herein, due to HCR, numerous GOD was efficiently immobilizated on the sensing surface and exhibited excellent catalysis towards glucose to in situ generate amounts of hydrogen peroxide (H2O2), which acted as luminol's co-reactant to significantly enhance the ECL signal. The proposed ECL immunosensor presented predominate stability and high sensibility for determination of IgG in the range from 0.1 pg mL(-1) to 100 ng mL(-1) with a detection limit of 33 fg mL(-1) (S/N=3). Additionally, the designed ECL immunosensor exhibited a promising application for other protein detection.

A homogeneous hemin/G-quadruplex DNAzyme based turn-on chemiluminescence aptasensor for interferon-gamma detection via in-situ assembly of luminol functionalized gold nanoparticles, deoxyribonucleic acid, interferon-gamma and hemin.

A homogeneous hemin/G-quadruplex DNAzyme (HGDNAzyme) based turn-on chemiluminescence aptasensor for interferon-gamma (IFN-γ) detection is developed, via dynamic in-situ assembly of luminol functionalized gold nanoparticles (lum-AuNPs), DNA, IFN-γ and hemin. The G-quadruplex oligomer of the HGDNAzyme was split into two halves, which was connected with the complementary sequence of P1 (IFN-γ-binding aptamer) to form the oligonucleotide P2. P2 hybridized with IFN-γ-binding aptamer and meanwhile assembled onto lum-AuNPs through biotin-streptavidin specific interaction. When IFN-γ was recognized by aptamer, P2 was released into the solution. The two lateral portions of P2 combined with hemin to yield the catalytic hemin/G-quadruplex DNAzyme, which amplified the luminol oxidation for a turn-on chemiluminescence signaling. Based on this strategy, the homogeneous aptasensor enables the facile detection of IFN-γ in a range of 0.5-100 nM. Moreover, the aptasensor showed high sensitivity (0.4 nM) and satisfactory specificity, pointing to great potential applications in clinical analysis.

Detection of metal ions (Cu2+, Hg2+) and cocaine by using ligation DNAzyme machinery.

The Cu(2+)-dependent ligation DNAzyme is implemented as a biocatalyst for the colorimetric or chemiluminescence detection of Cu(2+) ions, Hg(2+) ions, or cocaine. These sensing platforms are based on the structural tailoring of the sequence of the Cu(2+)-dependent ligation DNAzyme for specific analytes. The tethering of a subunit of the hemin/G-quadruplex DNAzyme to the ligation DNAzyme sequence, and the incorporation of an imidazole-functionalized nucleic-acid sequence, which acts as a co-substrate for the ligation DNAzyme that is tethered to the complementary hemin/G-quadruplex subunit. In the presence of different analytes, Cu(2+) ions, Hg(2+) ions, or cocaine, the pretailored Cu(2+)-dependent ligation DNAzyme sequence stimulates the respective ligation process by combining the imidazole-functionalized co-substrate with the ligation DNAzyme sequence. These reactions lead to the self-assembly of stable hemin/G-quadruplex DNAzyme nanostructures that enable the colorimetric analysis of the substrate through the DNAzyme-catalyzed oxidation of 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), ABTS(2-), by H(2)O(2) into the colored product ABTS(·-), or the chemiluminescence detection of the substrate through the DNAzyme-catalyzed oxidation of luminol by H(2)O(2). The detection limits for the sensing of Cu(2+) ions, Hg(2+) ions, and cocaine correspond to 1 nM, 10 nM and 2.5 µM, respectively. These different sensing platforms also reveal impressive selectivities.

Rapid determination of vitamin B12 concentration with a chemiluminescence lab on a chip.

This paper reports a novel method for the rapid determination of vitamin B(12) concentration in a continuous-flow lab-on-a-chip system. This new method is based on luminol-peroxide chemiluminescence (CL) assays for the detection of cobalt(II) ions in vitamin B(12) molecules. The lab-on-a-chip device consisted of two passive micromixers acting as microreactors and a double spiral microchannel network serving as an optical detection region. This system could operate in two modes. In the first mode, samples are acidified and evaluated directly in the microchip. In the second mode, samples are treated externally by acidification prior to detection in the microchip. In the first mode, the linear range obtained was between 1.00 ng ml(-1) to 10 µg ml(-1), R(2) = 0.996, with a relative standard deviation (RSD) of 1.23 to 2.31% (n = 5) and a limit of detection (lod) of 0.368 pg ml(-1). The minimum sample volume required and the analytical time were 30 µl and 3.6 s, respectively. In the second mode, the linear range obtained was between 0.10 ng ml(-1) to 10 µg ml(-1), R(2) = 0.994, with the RSD of 0.90 to 2.32% (n = 6) and a lod of 0.576 pg ml(-1). The minimum sample and the analytical time required were 50 µl and 6 s, respectively. The lab on a chip working in mode II was successfully used for the determination of vitamin B(12) concentrations in nutritional supplemental tablets and hen egg yolks.

Application of optimized chemiluminescence assay for determination of the antioxidant capacity of herbal extracts.

A chemiluminescence (CL) assay for the determination of antioxidant capacity (AOC) has been optimized and applied to analyses of herbal extracts in the present study. The optimal concentrations of reagents (luminol, H2O2, horseradish peroxidase) have been determined, as well as the optimal reaction conditions (wavelength, pH, temperature, sample volume). All of the measurements were performed at the emission maximum of the oxidized form of luminol (425 nm). The optimal concentrations of the reagents were determined as follows: 1.6 mmol/L luminol, 7.5 mmol/L H2O2 and 0.14 U/mL horseradish peroxidase activity in the reaction mixture. Analyses were carried out in phosphate buffer, pH 7.4, at room temperature. With the optimized CL assay, the AOCs of various water and methanol herbal extracts were determined (dog rose hips, plantain leaves and coltsfoot and thyme flowers) and the results were compared to those obtained by other classical methods for the evaluation of antioxidants. Strong correlations (r > 0.9) with the Folin-Ciocalteau assay and the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH)(●) assay are confirmed, although there is no correlation between AOC and the concentration of ascorbic acid in the samples analysed. This optimized CL assay is simple, rapid and reliable, and it represents a good alternative to classical methods (Folin-Ciocalteau, DPPH(●)) for the determination of AOC of herbal extracts and other food samples.

[Chemiluminescence analysis of oil oxidizing bacteria Actinetobacter calcoaceticus extracts: effects of the extracts on pSoxS-lux biosensor].

A comparative H2O2-luminol- and Fe(II)-induced chemiluminescence analysis of extracts of two strains of marine oil oxidizing bacteria Actinetobacter calcoaceticus cultivated either in the presence or absence of oil was carried out. Effects of these extracts on E. coli MG1655 biosensor (pSoxS-lux) were studied. Activation of H2O2-induced chemiluminescence in the presence of oil was observed. This suggests activation of free radical lipid peroxidation. Aqueous extracts of microorganisms cultivated in the presence of oil were shown to activate reactive oxygen species production (ROS) in Fe(II)-induced chemiluminescence reaction mixture. Acetone-ethanol extracts induced antioxidative systems of both strains. Chemiluminescence analysis in a biological system carried utilizing E. coli MG1655 (pSoxS-lux) revealed that aqueous extracts of the strains cultivated in the absence of oil contained potential antioxidants.