QUANTUM YIELDS OF THE LUMINOL CHEMILUMINESCENCE REACTION IN AQUEOUS AND APROTIC SOLVENTS*

Abstract— Quantum yields for luminol (3-aminophthalic hydrazide) chemiluminescence reactions fall into two classes depending on oxidizing conditions. In aprotic solvents the quantum yield is high and the excitation yield which allows for the fluorescence quantum yield of the product, is 0·09 and is unaffected by changes in solution temperature or polarity, or the presence of quenchers. In aqueous solution under optimum pH conditions (11–13), hydrogen peroxide oxidation results in a high chemiluminescence quantum yield with an excitation yield of 0·04 again unaffected by temperature, viscosity or quenchers. Other oxidizing conditions produce lower quantum yields probably by the introduction of competing chemical pathways. The luminol chemiluminescence light standard has been used to calibrate a spectrofluoro-meter with results in good agreement with the quantum yields of the ferrioxalate actinometer and the fluorescence of quinine sulfate and diphenylanthracene.     

Influence of luminol on the chemiluminescence intensity in Fenton’s reaction

The kinetics of Fe⁺² oxidation and buildup of luminol oxidation products during Fenton’s reaction at pH 2 have been calculated. The characteristics of the process in neutral (pH 6) and alkaline (pH 12) media have been evaluated. The calculation results have been compared with experimental data on the yield of chemiluminescence induced by Fenton’s reagent and luminol. It has been shown that trivalent iron ions suppress the luminol emission. The presence of iron or another transition metal in the sample can significantly reduce the chemiluminescence quantum yield after luminol introduction if.     

Subnanogram determination of aniracetam in pharmaceutical preparations and biofluids by flow injection analysis with chemiluminescence detection based on its enhancement of the myoglobin-luminol reaction

A novel flow injection chemiluminescence method with a myoglobin-luminol system is described for determining aniracetam. Myoglobin-bound aniracetam produced a complex that catalyzed the chemiluminescence reaction between luminol and myoglobin, leading to fast chemiluminescence. The chemiluminescence intensity in the presence of aniracetam was remarkably enhanced compared with that in the absence of aniracetam. Under the optimum reaction conditions the chemiluminescence increment produced was proportional to the concentration of aniracetam in the range of 0.1-1000.0 ng/mL (R2 = 0.9992), with a detection limit of 0.03 ng/mL (3delta). At a flow rate of 2.0 mL/min, the whole process, including sampling and washing, could be completed in 0.5 min, offering a sampling efficiency of 120/h; the RSD was less than 3.0% (n = 5). The method was satisfactory for determination of aniracetam in pharmaceutical preparations and human urine and serum samples. A possible mechanism of the reaction is also discussed.     

THE REACTIVITY OF SUPEROXIDE (O2-) and ITS ABILITY TO INDUCE CHEMILUMINESCENCE WITH LUMINOL

Abstract— The quantum yield and the kinetics of O -induced luminol chemiluminescence was investigated in a broad pH interval at varying luminol and concentrations. It is suggested that the weak chemiluminescence observed is mediated via a luminol-superoxide-adduct proposed to be an a-hydroxyperoxyl radical. At pH 7 the maximum quantum yield of chemiluminescence per initial percent was determined to be 4 times 10^-8. The degree of involvement in phagocytosis and related processes should be viewed against this maximum limit.     

Flow-injection chemiluminescence determination of chlorinated isocyanuric acids

A rapid and sensitive flow-injection chemiluminescence method is described for the determination of dichloro- and trichloroisocyanuric acids based on the chemiluminescence produced during their reaction with luminol in alkaline medium. The effects of analytical and flow-injection variables on these chemiluminescence systems and determination of both oxidants are discussed. The optimized method yielded 3sigma detection limits of 8x10(-8) and 5x10(-8) mol L(-1) for the sodium dichloroisocyanurate and trichloroisocyanuric acid, respectively. The optimum conditions were found to be as follows: NaOH, 1x10(-1) mol L(-1); luminol, 5x10(-3) mol L(-1); KI, 2x10(-3) mol L(-1) and flow rate, 3.5 mL min(-1).     

Development of a quantitative bio/chemiluminescence spectrometer determining quantum yields: re-examination of the aqueous luminol chemiluminescence standard

We have developed a bio/chemiluminescence spectrometer with a cooled charge-coupled-device (CCD) detector to obtain a quantitative luminescence spectrum as the absolute number of all emitted photons at each wavelength. The integrated area of the spectrum divided by the number of reacted substrate molecules gives the quantum yield. Calibration of the absolute sensitivity of the CCD-spectrometer system was performed by using lasers and a tungsten lamp with calibrated powers as primary light standards, and calibration of the light-collection efficiency of the spectrometer with several kinds of cells for liquid samples was achieved by introducing a simple reference double-plate cell. The reference cell is not convenient for final bio/chemiluminescence measurements but is useful for the calibration because it has well-defined angular dependence of light emission, allowing accurate calculation of the light-collection efficiency. Using this CCD-spectrometer system, we re-examined the quantum yield of aqueous luminol chemiluminescence with H2O2 catalyzed by horseradish peroxidase. The quantum yield was constant for a wide range of luminol concentrations, whereas it changed and had an optimum against H2O2 concentrations. The optimum quantum yield was 1.23(+/-0.20)%, which is in good agreement with previously reported values.     

Investigations on the Chemiluminescence Reaction Between Luminol and Reductant

The chemiluminescence reaction between luminol and reductant was investigated by a flow-injection system. The results show that this reaction can be used for the determination of the reductant which is active to chemiluminescence in alkaline lumminol, and of the substance which can, in appropriate manner, be converted into the active reductant. The mechanism of the chemiluminescence reaction may be that the reductant reduces dissolved oxygen to O2 , then O2 reacts with alkaline luminol to generate chemiluminescence.     

Activity of Aliphatic Diperoxy Acids in the Luminol Chemiluminescence Reaction

The activity of dilute aqueous solutions of aliphatic diperoxysuccinic, -glutaric, -adipic, and -sebacic acids in the luminol chemiluminescence reaction is studied. This activity can be detected at a low (10^–6–10^–5M) concentration of luminol in weakly alkaline solutions; it increases as the alkyl chain length of the acid decreases. The best conditions for chemiluminescence have been selected. It has been shown that individual or total diperoxy acids can be determined by luminol chemiluminescence. The determination limit for diperoxyadipic acid used as an example has been estimated at 2.0 ng/mL.     

STUDIES ON THE CHEMILUMINESCENCE OF LUMINOL IN DIMETHYLSULFOXIDE AND DIMETHYLSULFOXIDE-WATER MIXTURES*

Abstract— The absorption, fluorescence, and chemiluminescence (CL) characteristics of luminol have been studied in basic dimethylsulfoxide (DMSO) and various basic DMSO — water mixed solvents. It has been shown that the luminol dianion can be produced quantitatively in carefully deoxygenated ‘dry’ DMSO using potassium t-butyl alcoholate (BTO) as the base. A direct correlation has been found between the intensity of CL and the concentration of luminol dianion, Indicating that the dianion is the reactive species in the chemiluminescent reaction in DMSO. Increasing concentrations of water in the mixed solvents greatly reduced the CL intensity because of the decrease in luminol dianion concentration. Solvent effects on the fluorescence of 3-aminophthalic acid samples was noted and identified. Chemiluminescent spectra of luminol were run as a function of solvent composition, and compared with fluorescence spectra in the same media. The need for correcting these spectra for comparison was noted. Stopped flow kinetic studies were run in DMSO and DMSO-water mixtures. These data can be interpreted on the basis of two second-order steps and one first-order step in the reaction. The first-order step is probably last and probably arises from decomposition of a ‘peroxy’ intermediate. The rate constant for this step is k= 1·2±0·3×10^-1 sec^-1.     

Gold Nanoparticles‐enhanced Chemiluminescence Determination of Fenfluramine

A simple and sensitive chemiluminescence method was developed for the determination of fenfluramine. The chemiluminescence signal arising from the reaction between alkaline luminol and N‐bromosuccinimide was found to be greatly enhanced by fenfluramine in the presence of gold nanoparticles. But fenfluramine alone slightly inhibited the CL signal of N‐bromosuccinimide‐luminol in the absence of gold nanoparticles. The experimental parameters that affected the chemiluminescence signal were thoroughly investigated. Under the optimum experimental conditions, the enhanced chemiluminescence intensity was proportional to the concentration of fenfluramine in the range of 0.005‐1.0 mg/L. The detection limit was 0.9 μg/L fenfluramine with a relative standard deviation of 2.5% for 0.1 mg/L fenfluramine solution (n = 11). The method was applied to the determination of fenfluramine in some weight‐reducing tonics and in spiked human urine. A possible CL reaction mechanism was proposed.     

反相流动注射胶束增敏化学发光分析法测定硝苯地平

基于胶束介质中硝苯地平对碱性鲁米诺-过氧化氢化学发光体系的增敏作用,结合反相流动注射技术,建立了流动注射化学发光分析法测定硝苯地平的新方法.硝苯地平浓度在3.5×10-10～4.0×10-8 g·mL-1范围内时,化学发光强度与硝苯地平的浓度呈良好的线性关系,其相对标准偏差为1.4%(n=11,c=3.5×10-9 g...     

Solvent Effect on the Chemiluminescence of Acridinium Thioester: A Computational Study

Chemiluminescent labelling, which is one of the promising procedures of modern immunodiagnostics, is increasingly carried out using acridinium derivatives, an oxidant, and an alkaline aqueous environment. However, the efficiency of the chemiluminescence of luminol or acridinium esters is higher in non-aqueous solvents such as dimethyl sulfoxide or acetonitrile. Therefore, the search for a new environment for the chemiluminescence reaction, especially the one characterized by a higher quantum yield of chemiluminescence, is one of the aims of current research. Using computational methods (DFT and TD DFT with PCM model of solvent), we examined thermodynamic and kinetic data concerning the chemiluminescence and competitive dark pathways. Our results suggest that better characteristics of the chemiluminescence reaction of acridinium thioester are observed in nonpolar solvents, such as methylcyclohexane, n-hexane and n-pentane, than in aqueous media used so far. Further experimental verification is necessary to confirm the possible application of proposed nonpolar solvents in chemiluminescent labelling and hence in immunodiagnostics.     

Chemiluminescence biosensor chip based on a microreactor using carrier air flow for determination of uric acid in human serum

A chemiluminescence biosensor on a chip coupled to a microfluidic system and a microreactor is described in this paper. The chemiluminescence biosensor measured 25 x 75 x 6.5 mm in dimension, and was readily produced in an analytical laboratory. The sol-gel method is introduced to co-immobilize horseradish peroxidase (HRP) and luminol in the microreactor, and to immobilize uricase in the enzymatic reactor. The main characteristic of the biosensor was to introduce air as the carrier flow instead of the more common solution carrier for the first time. The uric acid was determined by a chemiluminescent (CL) reaction between the hydrogen peroxide produced from the enzymatic reactor and luminol under the catalysis of HRP in the microreactor. The linear range of the uric acid concentration was 1 to 100 mg L(-1) and the detection limit was 0.1 mg L(-1) (3sigma).     

CdTe量子点增敏鲁米诺-KMnO4化学发光对间苯二酚的测定

碱性介质中,CdTe量子点能够强烈地增强鲁米诺-KMnO4体系的化学发光,间苯二酚对该体系的化学发光有很强的抑制作用.该文结合流动注射分析法,建立了测定间苯二酚的新方法,并对可能的反应机理进行了探讨.结果表明,在优化实验条件下,间苯二酚在1.0×10-9 ～5.0×10-5 mol/L的浓度范围内与发光强度呈良好的线性关系,检出限(S/N=3)为8.0×10-10 mol/L.对于1.0×10-7 mol/L间苯二酚,测定11次的相对标准偏差为2.6%.将该体系用于水样中间苯二酚的测定,回收率为96% ～104%,相对标准偏差为1.9% ～3.3%.     

Investigations of the Enhancer Effect of a High-Salt Concentration Medium on the Luminol Chemiluminescent Reaction

The enhancer effect of a high-salt concentration medium on the luminol chemiluminescence reaction catalyzed by either soluble or immobilized peroxidase has been investigated. Some widely used salts were tested at high concentration (up to 3 mol L): potassium chloride, sodium chloride, ammonium sulfate and calcium chloride. The magnitude of the light intensity enhancement depends on the nature of the salt and on the form of the peroxidase, i.e. free in solution or immobilized. The enhancement is observed whether the catalyst of the chemiluminescence reaction is peroxidase or ferricyanide. Both the enhanced or nonenhanced luminol chemiluminescence spectra have a maximum wavelength emission at around 425 nm. The results reported in this study are in favor of an action of the salts on the nonenzymatic steps of the luminol chemiluminescence reaction rather than a modification of the enzymatic process.     

An ultrasensitive and highly selective determination method for quinones by high-performance liquid chromatography with photochemically initiated luminol chemiluminescence

Quinones are a class of compounds of substantial toxicological and pharmacological interest. An ultrasensitive and highly selective chemiluminescence (CL) method was newly developed for the determination of quinones based on the utility of photochemically initiated luminol CL. The method involved ultraviolet (UV) irradiation of quinones to generate reactive oxygen species (ROS) through the unique photosensitization reaction accompanied with the photolytical generation of 3,6-dihydroxyphthalic acid (DHPA) from quinones. The photoproducts were detected by luminol CL reaction. Interestingly, it was noticed that DHPA had enhancement effect for the luminol CL. The generation of the enhancer (DHPA) in association with the oxidant (ROS) in the photochemical reaction greatly increases the sensitivity and selectivity of the proposed luminol CL method. In order to elucidate the type of ROS produced by the photosensitizaion reaction in relation to the proposed CL reaction, we investigated the quenching effect of selective ROS scavengers in the luminol CL. Although several ROS were generated, superoxide anion was the most effective ROS for the generated CL. Moreover, the enhancement mechanism of DHPA for luminol CL was confirmed. The enhancement was found to be through the formation of stabilized semiquinone anion radical that provided long-lived CL. The generation of the semiquinone radical was confirmed by electron spin resonance technique. Furthermore, we developed an HPLC method with on-line photochemical reaction followed by the proposed CL detection for the determination of four quinones. A luminol analogue, L-012, was used for its high sensitivity. The detection limits for quinones obtained with the proposed method (S/N = 3) were in the range 1.5–24 fmol that were 10–1000 times more sensitive compared with the previous methods. Finally, the developed HPLC-CL system was successfully applied for the determination of quinones in airborne particulate samples collected at Nagasaki city.     

Determination of ATP via the photochemical generation of hydrogen peroxide using flow injection luminol chemiluminescence detection

The determination of ATP using the coupling between a photochemical reaction and a chemiluminescence reaction in a flow injection (FI) system is described. The method is based on the reaction of glucose with ATP catalyzed by hexokinase and Mg²⁺ ions. The glucose that is not consumed by ATP is subsequently photooxidized using 9,10-anthraquinone-2,6-disulfonate as a sensitizer. The hydrogen peroxide produced in the photochemical reaction is monitored through the chemiluminescence reaction with luminol catalyzed by hematine. There is a linear relationship between the decrease in the chemiluminescence response and the ATP concentration at a constant concentration of glucose. Under the optimum conditions, the calibration graph is linear in the range 0.20–50.5 mg L^–1 with a throughput of 25 samples per hour and relative standard deviations between ±0.62 and ±1.42%. The limit of detection is 0.07 mg L^–1. The method was used for the determination of ATP in pharmaceuticals, milk, and soils.     

Apparatus for mechanistic and analytical studies of the electrogenerated chemiluminescence of luminol

A new apparatus based on the rotating ring—disc electrode system is described. The symmetric double-step potential is connected to the ring electrode to oxidize luminol, while the disc electrode is maintained at a negative potential to reduce oxygen to hydrogen peroxide. Because of the electrode rotation, hydrogen peroxide is immediately transported to the ring electrode at which it reacts with luminol oxidation product to emit light. Preliminary electrogenerated chemiluminescence measurements indicate that the intensity of the chemiluminescence of luminol is highly dependent on the ring and disc electrode materials and that some metal ions have a catalytic or inhibitive effect on this luminescence reaction of luminol.     

Efficiency of Electron Transfer Initiated Chemiluminescence

Although the mechanisms of many chemiluminescence (CL) reactions have been intensively studied, no general model has been suggested to rationalize the efficiency of these transformations. To contribute to this task, we report here quantum yields for some well‐characterized CL reactions, concentrating on recent reports of efficient transformations. Initially, a short review on the most important general CL mechanisms is given, including unimolecular peroxide decomposition, electrogenerated CL, as well as the intermolecular and intramolecular catalyzed decomposition of peroxides. Thereafter, quantum yield values for several CL transformations are compiled, including the unimolecular decomposition of 1,2‐dioxetanes and 1,2‐dioxetanones, the catalyzed decomposition of appropriate peroxides and the induced decomposition of properly substituted 1,2‐dioxetane derivatives. Finally, some representative examples of quantum yields for complex CL transformations, like luminol oxidation and the peroxyoxalate reaction, in different experimental conditions are given. This quantum yield compilation indicates that CL transformations involving electron transfer steps can occur with high efficiency in general only if the electron transfer is of intramolecular nature, with the intermolecular processes being commonly inefficient. A notable exception to this general rule is the peroxyoxalate reaction which, also constituting an example of an intermolecular electron transfer system, possesses very high quantum yields.     

流动注射纳米微反应器化学发光法测定盐酸阿米替林

在酸性条件下,盐酸阿米替林分子中氮原子被质子化后与阴离子AuCl4-形成离子缔合物,该缔合物被二氯甲烷带入鲁米诺的氯化十六烷基三甲基铵反胶束纳米微反应器中,离解出来的AuCl4-立即与鲁米诺产生化学发光.在一定浓度范围内,发光强度与盐酸阿米替林的含量成线性关系,从而间接测定盐酸阿米替林的含量.在优化的实验条件下,线性范...