Please suggest me a standardized method to prepare DNS reagent for the determination of reducing sugar by using N acetyl D glucosamine.
Thank you
Dear Renuka,
Preparation steps of 3,5-dinitrosalicylic acid solution 100(ml):
1. Dissolve 30(g) of Potassium sodium tartrate tetrahydrate in 20(mL) distilled water. Add the salt to water gradually.
2. Prepare 2(N) sodium hydroxide solution 20 (mL). Dissolve smoothly sodium hydroxide in water using a glass rod and stirring the mixture of granules in water.
3. Dissolve 1(g) of 3,5-dinitrosalicylic acid in 50 (mL) of distilled water while the solution is mixed by magnetic stirrer with hot plate at 90-95˚C.
4. Add gradually the solution of potassium sodium tartrate tetrahydrate (prepared solution in Step 1) to 3,5-dinitrosalicylic acid solution (prepared solution the Step 3) while the solution is mixed by magnetic stirrer with hot plate at 90-95˚C.
5. Add slowly 2(N) sodium hydroxide solution (the solution prepared in step 2) to the solution prepared in step 4 while the solution is mixed by magnetic stirrer with hot plate at 90-95˚C.
6. After the components are completely dissolved, filter the final solution by filter paper.
7. Transfer the solution in dark glass bottles and storage at ambient temperature.
good luck!
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