Moreover, the anthrone method produces a bluish-green coloured complex while the DNSA method produces a reddish-brown coloured complex. In this experiment, DNS method will be used. Mix: Distilled Water 1416 ml. Using twelve commercial enzyme preparations, the comparison of the NS and DNSassays in determination of cellulase, -glucanase, xylanase, and -mannanase activities was carried out. Previous question Next question Get more help from Chegg. DNS method The DNS method for estimating the concentration of reducing sugars in a sample Reducing sugars contain free carbonyl group, have the property to reduce many of the reagents. We suggest that the DNSA (3,5-dinitrosalicylic acid) test, a quantitative measure of reducing sugars, is used in this context. Stop the reaction by addition of 1 ml of DNS reagent mix well and keep the test tubes in a boiling water bath for 10 minutes. Post-16 Biology specifications in England require students to use ‘appropriate instrumentation to record quantitative measurements, such as a colorimeter ...’. Constructing a standard curve / graph for maltose helps us to estimate concentration of reducing sugars present in an unknown sample and for determining the activity of amylase enzyme in forthcoming experiments.The standard curve for maltose is usually constructed using 3, 5-Dinitro salicylic acid (DNS) as the reagent. Prepare by dissolving 1.0 gm of 3,5-dinitrosalicylic acid in 50 ml of reagent grade water. Ensure that the bottle is closed tightly and swirl to mix the contents. Preparation of Reagents: 3,5-dinitrosalicylic acid [DNS]: About 1g of DNS is dissolved in 50ml of distilled water. •The dinitrosalicylic acid (DNS) method for estimating the concentration of reducing sugars in a sample. IMPORTANTthe Safety Data Sheet supplied with the product refers to the DNSA reagent base before you have added sodium hydroxide to it. (ii) Working standard sodium: Take 10 mL from this stock solution and make up the volume to 100 mL. 3,5-Dinitrosalicylic acid (DNS or DNSA, IUPAC name 2-hydroxy-3,5-dinitrobenzoic acid) is an aromatic compound that reacts with reducing sugars and other reducing molecules to form 3-amino-5-nitrosalicylic acid, which strongly absorbs light at 540 nm. The colour of the reagent changes from yellow to orange or red, depending upon the concentration of reducing sugar present. All of the prices on this page are in GBP and do not include Value Added Tax (VAT). Mix well. The Nelson-Somogyi (NS) and 3,5-dinitrosalicylic acid (DNS) assays forreducing sugars are widely used in measurements of carbohydrase activities against differentpolysaccharides. 0.02 M Sodium phosphate buffer, pH 6.9 with 0.006 M sodium chloride; 2 N Sodium hydroxide; Dinitrosalicylic acid color reagent. Pages 16 This preview shows page 8 - 13 out of 16 pages. A typical method to make it would be: Slowly add 10.6 grams of 3,5 - Dinitrosalicylic acid and 19.8 grams of Sodium hydroxide to 1.416 liters of distilled or deionized water. Once the sodium hydroxide has been added, the concentration of sodium hydroxide in the complete DNSA reagent is 0.4 M. This article is cited by 15145 publications. Operations Management. This method tests for the presence of free carbonyl group (C=O), the so-called reducing sugars. Constructing a standard curve / graph for maltose helps us to estimate concentration of reducing sugars present in an unknown sample and for determining the activity of amylase enzyme in forthcoming experiments.The standard curve for maltose is usually constructed using 3, 5-Dinitro salicylic acid (DNS) as the reagent. Prepare by dissolving 1.0 gm of 3,5-dinitrosalicylic acid in 50 ml of reagent grade water. The colour of the liquid will change from opaque yellow to clear, bright orange. Add 1 ml of a 40% potassium sodium tartrate (Rochelle salt) solution to stabilize the color. Dip the test strip into the liquid. Generally, Anthorne method is a qualitative method while the DNSA method is a quantitative method. Reagents Required. It was also used to measure the effects of silver nanoparticles on the membrane leakage of the reducing sugars. Protect from carbon dioxide and store no longer than 2 weeks. Thus it helps to meet two of the important practical requirements of the current (English) biology specifications. Method Unlike other carbohydrates, sucrose is the only non-reducing common disaccharide. Keep in boiling water bath for 15 minutes. •Reducing sugars contain free carbonyl group, have the property to many of the reagents. 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