DAOS, one of the new Trinder's reagents, the full Chinese name is N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt, commonly used in uric acid and renal function detection The detection kit has the advantages of good water solubility, high sensitivity, and strong stability. It is a white or light blue powder with CAS number 82692-97-5. This product is sensitive to light and humidity.
Features: As a member of the new Trinder's reagent, DAOS has high water solubility compared with other chromogenic substrates. It is a stable aniline analogue. The pH range of the chromogenic process and oxidation reaction is wide, and it is widely used Diagnostic testing and biochemical tests. It has several advantages over conventional color-producing reagents in the colorimetric determination of hydrogen peroxide activity. The new Trinder’s reagent is stable enough to be used both in solution and in test line detection systems.
Preparation of DAOS detection solution:
1. Dissolve 23 mg DAOS in 10 ml PBS buffer to prepare a 6.6 mM DAOS solution. (The specific solubility can be adjusted as needed)
2. Dissolve 14 mg 4-aminoantipyrine (4-AA) with 10 ml PBS buffer to prepare a 6.6 mM 4-AA solution.
3. Prepare 2 U/ml horseradish peroxidase solution with PBS.
4. Mix equal volumes of each solution to prepare the test solution. Store the detection solution in the dark at 4°C.
Detection steps:
1. Prepare sample solutions for enzymatic oxidation reactions. The pH range of the buffer should be 5.5-9.5.
2. Use the same buffer to prepare a standard solution containing a known amount of substrate.
3. Add the appropriate unit of oxidase to the sample solution, and then add an equal volume of the detection solution.
4. Incubate the mixture at room temperature or 37°C for 30 minutes to 1 hour.
5. Determine the O.D. value at 593 nm.
6. Prepare a standard curve and determine the concentration of the substrate in the sample solution.
Detection principle: In the presence of hydrogen peroxide and peroxidase, the new Trinder's reagent is oxidatively coupled with 4-aminoantipyrine (4-AA) or 3-methylbenzothiazole sulfone hydrazone (MBTH) During the process, a very stable purple or blue dye is formed. The molar absorbance of the dye coupled with MBTH is 1.5-2 times higher than that of the dye coupled with 4-AA. The substrate is enzymatically oxidized by its oxidase to produce hydrogen peroxide, and the concentration of hydrogen peroxide corresponds to the concentration of the substrate. The amount of the substrate can be determined by the color development of the oxidative coupling reaction.
Precautions:
1. Sub-packaging: When taking a small amount of mg DAOS, the product needs to be sub-packed into the quantity needed each time to reduce the number of bottle openings and prevent the product from absorbing moisture.
2. Use: When using the product, take out the product from the freezer half an hour in advance and place it at room temperature. If there is any remaining after use, store the remaining DAOS in a sealed and light-proof container to avoid quality problems such as color or property changes. .
3. Preservation: Place DAOS in a 0-5 degree freezer, and record the time and batch number.
4. Transportation: Put the packed product with ice in the foam box and wrap the product with foam glue. Take care to avoid the water from the ice cubes to wet the product (we put two more if the temperature is high, and the temperature can be changed in winter. To decide)
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