Differences in the application of various chemiluminescent reagents such as luminol acridine ester
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There are mainly three types of chemiluminescent reagents: luminol derivatives, acridine esters, and electrochemiluminescence. The development of immunology in our country has gone through three main stages: Radioimmunoassay (RIA) is now in a declining phase, but it is still widely used in hospitals at the county level and above, with standardized reagent sets available ； Enzyme-linked immunosorbent assay (ELISA) is widely used in clinical settings; the technology is mature, but the reagents have not yet been standardized (methods that utilize chromogens such as ADPS, TOOS, MAOS, etc., in combination with hydrogen peroxide coupling fall into this category) ； Chemiluminescent immunoassay (CLIA) is a relatively advanced method that is used in some larger hospitals. It is already well-established abroad, but in China it is still in the introduction or development phase, relying mainly on imports. The chemiluminescent detection of luminol, isoluminol, and alkaline phosphatase AMPPD requires the involvement of enzymes; it belongs to the class of enzyme-catalyzed chemiluminescence reactions involving acridine esters such as acridinium salts, acridinesulfonamides, and acridinoylhydrazides. In contrast, luminescence reactions that do not require enzymes fall under direct chemiluminescence, offering higher sensitivity. Ruthenium tris(2,2-dipyrrolyl)picolate belongs to the category of electrochemiluminescence; this method demands high standards from testing facilities and involves expensive equipment, so it is rarely used in China. Here are some of the parameters; the differences in the application of various luminescent reagents require further discussion, or guidance from experts in this field. 1# Acridine ester DMAE-NHS 【English name】6’-Dimethyl-4’-(N-succinimidyloxycarbonyl)phenyl-10-methyl-acridinium-9-carboxylate methosulfate 【Chinese name】Acridine ester DMAE-NHS 9-(1-pyrrolidinyl)oxycarbonyl]-2,6-bis(oxocarbonyl)-10-methyl-acridinium methanesulfonate 【CAS#】115853-74-2 【Molecular weight】594.589 【Appearance】Yellow solid/powder 【Molecular formula】C29H26N2O10S file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml1020\wps1.jpg 【Appearance】Yellow solid or powder 【Storage conditions】Room temperature, protected from light and moisture 【Uses】Used in chemiluminescence and immunological analysis, receptor analysis, as well as in research related to nucleic acids and peptides. As a chemiluminescent reagent for direct chemiluminescent immunoassay, it is used for the detection and analysis of antigens, antibodies, proteins, etc. No additional catalyst is required during the detection and analysis process. When DMAE-NHS is oxidized by H2O2 in an alkaline medium, it goes through a dioxygen intermediate to produce electropically excited N-methylacridinium ions; when these ions return to their ground state, they emit photons at 430 nm. 【Product Advantages】Purity ≥98%, good water solubility, stable manufacturing process, ensuring that the product appears as pure white crystal powder.
Luminol reagent 【English name】3-Aminophthalhydrazide 【Chinese name】Luminol/3-Aminobenzoylhydrazide/Luminescent ammonia 【CAS#】521-31-3 【Molecular weight】177.16 【Molecular formula】C8H7N3O2 【Storage conditions】Room temperature, protected from light and moisture 【Chemical properties】Soluble in alkaline solutions, soluble in dilute acids, practically insoluble in water, and insoluble in alcohols. Neutral or slightly acidic solutions exhibit a strong bright blue fluorescence when exposed to ultraviolet light. 【Use】Chemiluminescent reagent, commonly used in chemiluminescent immunoassays, such as the detection of metal cations and blood components. 【Luminescence rate detection】The optimal fluorescence wavelength is 425 nm (for detecting chemiluminescence rate in a solution of 60 mM K2S2O8, 100 mM K2CO3, and pH 11.5). Luminol/luminous ammonia/Luminol is one of the most commonly used reagents for liquid-phase chemiluminescence, owing to its simple structure, ease of synthesis, good water solubility, and high luminescent quantum efficiency. Since Albrecht first reported the chemiluminescent reaction between luminol and oxidants in alkaline solutions in 1928, research on this chemiluminescent system has been very active, leading to its application in many fields.
Allophorin chemiluminescent reagent 【English name】4-Aminophthalhydrazide 【Chinese name】4-Aminophthalhydrazide/6-Amino-2,3-dihydro-1,4-phthalazinedione 【CAS#】3682-14-2 【Molecular weight】177.16 【Molecular formula】C8H7N3O2 【Storage conditions】Room temperature, protected from light and moisture 【Appearance】Off-white powder 【Uses】Chemiluminescent reagent, commonly used in chemiluminescent immunoassays such as the detection of metal ions and blood components. 【Product Advantages】Purity: ≥98% (HPLC); good water solubility; stable manufacturing process; minimal batch-to-batch variation. Luminol and iso-luminol are both chemiluminescent agents that, under the catalysis of peroxidase or hemoglobin, iron, etc., react with hydrogen peroxide to emit blue fluorescence. Although heterorhodamine is not used as widely as rhodamine, it plays an irreplaceable role in blood tests. The cyclic hydrazide structure is the key structure responsible for chemiluminescence in luminol and iso-luminol compounds; modifications to this structure result in the near-complete loss of chemiluminescent properties ； Modifying the amino groups of luminol and isoluminol compounds results in a significant decrease in the luminescence efficiency of luminol, whereas the luminescence efficiency of isoluminol increases significantly.
2# Acridine Ester Me-DMAE-NHS 【English name】2’,6’-Dimethylcarbonylphenyl 10-Methyl-9-acridinecarboxylate 4’-NHS Ester Triflate 【Chinese name】Acridine Ester (Me-DMAE-NHS) 【Molecular weight】632.56 【Molecular formula】C29H23F3N2O9S 【Appearance】Yellow solid or powder 【Storage conditions】Room temperature, protected from light and moisture 【Uses】Can be used for labeling proteins, antigens, antibodies, nucleic acids (DNA, RNA), etc. 【Product Advantages】Purity: ≥98% (HPLC); good water solubility; stable manufacturing process; minimal batch-to-batch variation.
3# Acridine Ester NSP-DMAE-NHS CAS 194357-64-7: [English name] NSP-DMAE-NHS [Chinese name] Acridine Ester NSP-DMAE-NHS [CAS#] 194357-64-7 [Molecular weight] 590.60 [Appearance] Yellow solid/powder [Molecular formula] C30H26N2O9S [Storage conditions] Room temperature, protected from light and moisture [Uses] Used in research areas such as chemiluminescence and immunological analysis, receptor analysis, as well as the detection of nucleic acids and peptides. 【Product Advantages】Purity: ≥98% (HPLC); good water solubility; stable manufacturing process; minimal batch-to-batch variation. 【Properties】Acridine esters are a class of chemicals that can be used as chemiluminescent labels. In an alkaline H2O2 solution, when the molecules of acridine esters are attacked by hydrogen peroxide ions, the substituents on the acridine ring can form unstable diethoxides with C-9 on the acridine ring and H2O2 (hydrogen peroxide). These diethoxides then decompose into CO2 and N-methylacridine in an electronically excited state. Desheng is a established chemical reagent company specialized in the research and development as well as production of blood test reagents. It boasts 14 years of experience in these areas. Luminescent reagent substrates have always been a key focus of the company’s research; the acridine ester series exhibits high luminescence efficiency and belongs to the category of directly luminescent substances. - This article is from **Chemical Engineering Forum; the original address is: https://bbs.mahoupao.com/thread-218882-1-1.html
This post was last edited by Desheng on 2020-9-9 at 17:14. 4# Acridine ester NSP-SA-NHS acridinium salt (NSP-SA-NHS) chemiluminescent substrate 【English name】 3-(((3-(N-succinimidyloxycarboxypropyl)sulfonyl)amine)carboxyl]-10-acridiniumyl)-1-propanesulfonate 【Chinese name】 Acridinium salt (NSP-SA-NHS) 【CAS#】 199293-83-9 【Molecular weight】 681.73 【Molecular formula】 C32H31N3O10S2 file:///C:\Users\ADMINI~1\AppData\Local\Temp\ksohtml10760\wps1.jpg 【Appearance】 Yellow solid or powder 【Storage conditions】 Room temperature, protected from light and moisture 【Uses】 Can be used for labeling proteins, antigens, antibodies, nucleic acids (DNA, RNA), etc. 【Product Advantages】Purity: ≥98% (HPLC); good water solubility; stable manufacturing process; minimal batch-to-batch variation. The related compounds have been proven to be very advantageous chemiluminescent markers; their stability, activity, and sensitivity exceed those of radioactive isotopes.
5# Acridinium Salt NSP-SA Chemiluminescent Substrate 【English Name】(Tosyl)carbamoyl)acridinium-10-yl)propane-1-sulfonate (NSP-SA) 【Chinese Name】Propanesulfonate acridinium salt (NSP-SA) 【CAS#】211106-69-3 【Molecular Weight】584.66 【Molecular Formula】C28H28N2O8S2 【Appearance】Yellow solid or powder 【Storage Conditions】Room temperature, protected from light and moisture 【Uses】Can be used for labeling proteins, antigens, antibodies, nucleic acids (DNA, RNA), etc. Dilute solutions of acridine and its salts exhibit purple or green fluorescence; the dilute solutions of the salts show green fluorescence. Upon further dilution, hydrolysis of the salts leads to the formation of free acridine, which then emits purple fluorescence. Acridine salts have a wide range of applications, including use as photocatalysts and in the preparation of dyes. 【Product Advantages】Purity: ≥98% (HPLC); good water solubility; stable manufacturing process; minimal batch-to-batch variation. - This article is from **Chemical Engineering Forum; the original address is: https://bbs.mahoupao.com/thread-218882-1-1.html
6# Acridine ester, Acridinyl hydrazide NSP-SA-ADH 【English name】NSP-SA-ADH 【Chinese name】Acridinyl hydrazide (NSP-SA-ADH) 【Molecular weight】740.85 【Molecular formula】C34H40N6O9S2 【Appearance】Yellow solid or powder 【Storage conditions】Room temperature, protected from light and moisture 【Uses】Can be used for labeling proteins, antigens, antibodies, nucleic acids (DNA, RNA), etc. Acridhydrazide ends can be used to react with polysaccharides, nucleic acids, or proteins containing aldehyde groups, thereby acridine-labeling them. 【Product Advantages】Purity: ≥98% (HPLC); good water solubility; stable manufacturing process; minimal batch-to-batch variation. These are all acridine esters; different types have varying functional groups. Some of them are acridine sulfonates, acridine carboxylic acids, acridine sulfamides, etc., and can be used for labeling various proteins, antibodies, as well as nucleic acids such as DNA or RNA. – This article is from **Chemical Engineering Forum**, original address: https://bbs.mahoupao.com/thread-218882-1-1.html
Acridine salt NSP-SA, which is an acridine ester of acridic acid, cannot be used to label protein antibodies directly; a condensing agent such as EDCI must be added in order to enable coupling reactions with proteins that contain amino groups.
Chemiluminescence detection is being increasingly used in clinical applications, such as the following related tests: thyroid hormone testing, reproductive hormone testing, and adrenal/pituitary hormone testing； Anemia factors, tumor markers, infectious diseases ； Diabetes, cardiovascular system, viral markers ； For topics related to the chemiluminescence platform, such as bone metabolism, allergic diseases, and therapeutic drug monitoring, you can consult the customer service at Selenite6 on the official website of Desheng Technology
Both luminol and isoluminol are chemiluminescent agents that, under the catalysis of peroxidase or hemoglobin, iron, etc., react with hydrogen peroxide to emit blue fluorescence. Therefore, luminol can be used to detect trace bloodstains, as well as copper, iron, and cyanide in cells.