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edta disodium salt dihydrate
|Melting point||300℃||Boiling point||633ºCat760mmHg|
Luminol is a commonly used Luminol-based chemiluminescent reagent. Its luminescence mechanism is the luminescence of an oxidation reaction. It is catalyzed by horseradish peroxidase under alkaline conditions and is oxidized by H2O2 to produce 4-aminophthalic The excited intermediate of the acid emits photons when it returns to the ground state.
This product is a chemiluminescent reagent, commonly used in chemiluminescent immunoassay, such as metal cation and blood detection.
Preparation of isoluminol
Method 1: A method for preparing a high-yield isoluminol luminescent reagent, including the following steps:
(1) Preparation of phthalimide: In the reactor, add 37.1g of phthalic anhydride and 9.0g of urea, and heat up. When the temperature rises to 120-133 ° C, keep the reaction for 1-45min. At room temperature, phthalimide products can be obtained;
(2) Preparation of 4-nitrophthalimide: take 100-140mL concentrated sulfuric acid, put it in a reaction bottle, add 24mL fuming nitric acid, put it in an ice bath, cool to 3-8 ℃, in Rotate at 500-1000r / min, add 20g phthalimide under stirring, continue to stir at 10-15 ℃ for 4h, then put it at room temperature for 12-18h, pour the reaction liquid into 0.45-0.8kg crushed ice In the process, suction filtration is performed at a pressure of 5-15kPa to obtain: a product;
(3) Preparation of 4-nitrophthaloyl hydrazide: In a container, add 12g of 4-nitrophthalimide, 4g of hydrazine hydrate, 80mL of water, and reflux at 115-130 ℃ for 1-2 hours To obtain an orange-red solution. After cooling, adjust the pH to 3 with glacial acetic acid to precipitate a large amount of precipitate. Wash the precipitate with dilute acetic acid, filter with suction, and dry to obtain a light yellow solid product;
(4) Preparation of 4-aminophthaloyl hydrazide (isoluminol): 12.6g of 4-nitrophthaloyl hydrazide was warmly dissolved in 125mL of 5% sodium hydroxide solution under continuous stirring Drop the solution into a pure clear solution containing 56g of stannous chloride concentrated hydrochloric acid, add 2-5g of nano-titanium dioxide to the solution, and react with ultraviolet light at 20-30 ° C for 10-15min to obtain a colorless clear solution , Cooled to 0-minus 5 ℃, a large amount of white was obtained: flocculent precipitate, filtered with suction, the precipitate was dissolved with 10% ammonia, the insolubles were filtered off, and the filtrate was acidified with 20% acetic acid to obtain a white precipitate, which was dissolved with 2% NaOH Filtration with suction, the filtrate is acidified with 20% acetic acid to obtain a white precipitate, suction filtration, and drying to obtain a white powdery solid is the product of isoluminol;
Method 2: The method of preparing isoluminol using phthaloyl hydrazide as raw material includes the following steps:
Step 1: Preparation of diacetylphthaloyl hydrazide
With stirring at room temperature, 60 g of dry phthaloyl hydrazide waste (47% purity) and 200 ml of acetic acid were added to a 500 ml three-necked flask, and the mixture was stirred well. 30g of sodium acetate was added thereto, stirred at room temperature for 30 minutes, then 40g of acetyl chloride was added dropwise thereto, and the dripping was completed within 30 minutes. Stir at room temperature for 1 hour, then warm to 70 ° C and continue the reaction for 5 hours. Spot the plate to confirm that phthaloyl hydrazide has been completely converted into the desired product, stop the reaction, and let the reaction solution naturally drop to room temperature. The reaction solution is directly processed without further treatment.
Step 2: Preparation of nitrosodiacetylphthaloyl hydrazide
At room temperature, 15g of sodium nitrite was added in batches to the reaction solution from the previous step. After the addition, 15g was stirred at room temperature for 3 hours. The plate was confirmed to confirm that diacetylphthaloyl hydrazide had been completely converted into the desired product. After stopping the reaction, the acetic acid was recovered by distillation under reduced pressure, and the remainder was the crude 4-nitrosodiacetylphthaloyl hydrazide. The crude product was used directly in the next reaction without further treatment.
Step 3: Preparation of nitrosophthaloyl hydrazide
With stirring at room temperature, add the crude 4-nitrosodiacetylphthaloyl hydrazide obtained in the previous step to a 500ml three-necked flask, add 200ml of 95v / v% ethanol to it, mix well, and add 10g hydrogen Sodium oxide. After the addition was completed, the reaction was refluxed for 5 hours. The plate was spotted to confirm that 4-nitrosodiacetylphthaloyl hydrazide had been completely converted into the desired product, the reaction was stopped, and the reaction solution was naturally lowered to room temperature. The reaction solution is directly processed without further treatment.
Step 4: Preparation of isoluminol
Under stirring at room temperature, 10 g of sodium thiosulfate was added to the reaction solution in the previous step at once, and the mixture was stirred at room temperature for 30 minutes after the addition. Then, the reaction was heated at reflux for 3 hours. The plate was spotted to confirm that 4-nitrosophthaloyl hydrazide had been completely converted into the desired product. The reaction was stopped, the reaction solution was naturally cooled to room temperature, and then filtered, the filter cake was collected. The filtrate was rotary evaporated to recover 95% ethanol. The residue was combined with the filter cake, added to 200ml of water, heated to 80 ° C, and stirred for 30 minutes. Some solids were still insoluble, and filtered while hot. Concentrated hydrochloric acid was added to the filtrate to adjust the pH to 9, and the crystal was cooled and crystallized naturally. Filtration yielded 26g of light yellow powder product.
Its purity is not less than 98% (HPLC), with good water solubility, stable process, small difference between batches.
Desheng is a veteran chemical reagent company specializing in R & D and production of blood test reagents. It has 14 years of experience in research and development and production. Mino is so extensive, but has an irreplaceable role in blood testing. Business friends who need Yi Lumino can contact 17321456337 (WeChat same number) directly, and look forward to your call!
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