I need the detail answers. thanks Biochemisty Experiment 3 EXPERIMENT 3 Soluble

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I need the detail answers. thanks

Biochemisty Experiment 3 EXPERIMENT 3 Soluble carbohydrate quantification applying Anthrone assay 1. Principle In the anthrone assay, carbohydrate is dehydrated by using concentrated sulfuric acid to form furfural, which in turn condenses with anthrone (10-keto-9,10 dihydroanthracene) to form bluish-green complex which can be measured colorimetrically at wavelength of 620-630 nm by spectrophotometer. This a rapid and convenient assay for determination of hexoses, aldopentose and hexuronic acid, either in the free from or in form of polysaccharide. In this assay, the reagent preparation does not require the addition of distilled water, and anthrone is directly dissolved at a 2% concentration in concentrated sulfuric acid. This acid is powerful dehydrating agent involved in dehydrating sugars leading to formation of furfural which condenses with anthrone to give the colored product. 1. When the dried samples are used, why we need to use the fewer amounts compared to 2. How long does Anthrone reagent can be kept 3. Why anthrone reagent needs to be kept cold 4.Why test tubes must be put in ice-water the raw ones for using? before using? beaker/container when adding Anthrone solu- Sugar + conc. H,so, Blue-green colored complex Furfural (Anthrone) The accuracy of the reaction is based on the cleanness of equipment, the purification of the reagent, especially for sulfuric acid and the constant temperature during the boiling time 2.2 Chemicals +Alcohol 90 C Alcohol 80°C +Anthrone reagent + 0.01% Glucose solution tion?

Explanation / Answer

1) because dried sample already contains no water so the weight will contain only carbohydrate

2) Can be stored at room temperature for 2-3 days

3) and 4) Anthrone reagent is prepared in concentrated sulfuric acid. when we add the reagents to acid it dissociate immediately and releases heat.It could generate enough heat to blow out of the flask .

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