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3 morpholinopropanesulfonic acid,
edta disodium salt dihydrate
Acridine ester DMAE-NHS chemiluminescent agent is used for chemiluminescence and immunoassay, receptor analysis, nucleic acid and peptide detection and other research. As a chemiluminescent agent for direct chemiluminescence immunoassay, it is used for the detection and analysis of antigens, antibodies, proteins, etc. No additional catalyst is needed in the detection and analysis process. When DMAE-NHS is oxidized by H2O2 in an alkaline medium, an intermediate of ketone dioxide is produced to produce electrically excited N-methylacridone, which returns to the ground state at 430nm Release photons.
The first discovered chemiluminescence phenomenon was found in living organisms, such as fireflies, now known as bioluminescence, which is named for the visible light emitted by living organisms. In the late 1980s, Dubois studied the hot and cold water extract of fluorescent beetlein and found that in the presence of oxygen, luminescence occurs when hot water extract and cold water extract are mixed. By the end of the 19th century, it was discovered that simple non-biological organic compounds can also produce chemiluminescence. In 1877, Radziszewski discovered that ruthenium (2,4,5-triphenylimidazole) is oxidized by an agent such as hydrogen peroxide in an alkaline medium to emit green light. This reagent is still widely used today.
In 1935, Gleu and Petsch  reported for the first time that the reaction of Glossy (N,N-dimethyldiazidin nitrate) with hydrogen peroxide can produce chemiluminescence. Many reductions are made in the determination of glucose. Applications have been obtained for the determination of substances. Since then, the discovery of the chemiluminescence phenomenon of acridinium ester derivatives, the synthesis of chemiluminescent labeling reagents and chemiluminescence immunoassay have become a hot research topic in the 1980s, which has promoted the application of such analytical methods in body composition analysis. However, since the light intensity of most chemiluminescence phenomena is very weak and fleeting, the progress of early chemiluminescence research has been slow, and almost no substantive application has been made. It was not until the 1960s that the detection of faint light that was difficult to test in the past became possible, and chemiluminescence entered the era of quantitative analysis. The rapid oxidation of carbohydrates with free radicals as the initial reactants, the emergence of new systems such as fluorescent dyes such as fluorescein and eosin, and chemiluminescence of peroxidic oxalates reacting with hydrogen peroxide. It has played an important role in the high sensitivity detection of catechins, amino acids, and steroids.
2 The basic principle of chemiluminescence reaction
Chemiluminescence is the light produced during a chemical reaction. Usually this process can be described as:
A + B → [I]*→ Product + Light (1.1)
Where [I]* is the excited state product formed by the reaction of reactants A and B. The material in the excited state is unstable and will quickly transition to a lower energy state (such as the ground state), while the energy is light (usually The form of visible light is released . The chemiluminescence reaction can be divided into two categories according to the manner in which the excited state product is produced: one is an excited state product directly formed by a chemical reaction of a reactant in the system; and the other is a system which is easy to receive energy. The fluorescent substance is converted into an excited state after obtaining the energy released by the chemical reaction. Chemiluminescence can be applied to analytical measurements because the intensity of chemiluminescence is related to the rate of chemiluminescence, so all factors that influence the rate of reaction can be used as a basis for establishing assays. That is, a chemiluminescence process also includes a process of chemiluminescence reaction. Therefore, the intensity of chemiluminescence (ICL) depends on the rate of the chemical reaction, the efficiency of the excited state product, and the luminous efficiency of the excited state material.
ICL= ФCLdc/dt = ФEXФEMdc/dt (1.2)
Where ICL is the intensity of chemiluminescence (number of photons emitted per second); dc/dt is the rate of chemical reaction (number of molecules per second); ФCL is the quantum yield of chemiluminescence (number of photons emitted by each molecule participating in the reaction) ФEX represents the excited state quantum yield (excited state produced by each molecule participating in the reaction); ФEM represents the luminescence quantum yield (the number of photons generated per excited state). For a certain chemiluminescence reaction, ФCL is a certain value, but the measurement of chemiluminescence is susceptible to chemical reaction conditions such as pH, ionic strength, solution composition, temperature, etc., factors affecting the chemical reaction rate or any quantum efficiency. Both will change the luminous intensity. Therefore, the concentration of a substance in the reaction system can be determined by measuring the intensity of chemiluminescence under certain chemical reaction conditions. Since ICL = Ф CLdc / dt is integrated over time, ICL = ФCLc is obtained, and the luminescence intensity is proportional to the concentration of the reactant or product.
3 acridine chemiluminescence system
Lucigenin (N, N-dimethyldiazetidine nitrate) is one of the acridine compounds and one of the most widely studied and widely used luminescent reagents. It was first discovered by Glen and Petscsh in 1935. . Li Guanghao  studied the dynamic properties, photoluminescence spectroscopy, chemiluminescence spectroscopy and chemiluminescence mechanism of luster chemiluminescence system. This reaction is a fast kinetic reaction and is particularly suitable for post-column detection by capillary electrophoresis or chromatography. Among them, the most studied one is the acridinium ester compound. Under alkaline conditions, the acridinium ester is hydrolyzed by hydrogen peroxide to produce chemiluminescence. McCapra et al. conducted a detailed study on the chemiluminescence mechanism of acridine derivatives [6-8].
Generally, the chemiluminescence lifetime of acridine derivatives is quite short, but the modification of the modified acridine ring and the leaving group accelerates or retards this rapid kinetic process. Ruberto et al  introduced the reaction into capillary electrophoresis while isolating four different substituents of acridinium ester. In the detection of biologically active substances, research on this system has also been reported. Adrenalin and isoproterenol were measured under alkaline conditions by PL Wintrod and GIMakhatadze et al. . Methods for the determination of Vc using the Fe3+-Lucigenin luminescence system have also been reported . There are also a variety of drugs and biologically active substances, such as isoproterenol , benzenetriol , canamycin , ascorbic acid [15,16], etc. have achieved satisfactory results. .
The chemiluminescence quantum yield of acridinium ester is higher than that of luminol, and the acridine ester labeling condition is mild, the labeling rate is high, and the labeling does not affect the separation, so it has broad application prospects, often as chemiluminescence immunoassay and DNA luminescence detection. The chemiluminescent label of the needle is widely used for sensitive detection and diagnosis of various diseases, and can also be used for separation detection of amino compounds containing proteins, nucleic acids, peptides and the like.
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