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    Home > Biochemistry News > Biotechnology News > DNS (two nitrosic acid) color reduction sugar content determination experiment.

    DNS (two nitrosic acid) color reduction sugar content determination experiment.

    • Last Update: 2020-10-23
    • Source: Internet
    • Author: User
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    related topics .The determination experiment of reduced sugar is the basic experiment in
    bio-chemical
    technology, and it is the experimental operation technique that must be mastered in the bio-chemical course. In this paper, the experimental principles, steps, preparation of DNS
    reagents
    , the drawing of glucose standard curves, and the determination of samples are listed.I. Experimental purpose, DNS (denitrification salyanic acid) color reduction sugar content reduction experimental purpose to learn and understand the basic principles of reduced sugar and total sugar determination, color reduction sugar measurement operation steps and
    -dphotometer
    use method., experimental principle, the determinationreduction sugar is the basic method of sugar quantitative determination.is a sugar that contains free aldehyde or ketone. Monosaccharies are reduced sugars, double sugars and polysaccharies are not necessarily reduced sugars, such as lactose and maltose are reduced sugars, sucrose and starch are non-reducing sugars. Using the different solubility of various sugars,
    single
    , double sugar and polysaccharides in the sample can be extracted separately. For non-reductive double sugar and polysaccharides, the degradation of acid hydrolyzed into reduced monosaccharides can be determined, and then the content of reduced sugar and total sugar in the sample (often measured in glucose content) was calculated separately.dishing sugar under alkaline conditions
    heating
    can be oxidized into glycoic acid and other products, while oxidant 3,5-nitrosic acid is reduced to brown-red 3-amino-5-nitrate salginic acid. Within a certain range, the amount of reduced sugar is directly related to the depth and light of the brown-red substance color, and the light density value is measured at 540nm wavelength using a 12guangometer, the standard curve is checked and calculated, the content of reduced sugar and total sugar in the sample can be found. Since polysaccharide hydrolytic monosaccharides, each break of a glycoside bond needs to be added to a
    molecule
    water, so the polysaccharide content should be calculated multiplied by a factor of 0.9., the preparation of DNS reagentsadds 6.3 grams of DNS and 262 ml of 2mol/L sodium hydroxide to 500 ml of hot water solution containing 182 grams of potassium saltate In the liquid, plus 5 grams of heavy phenol and 5 grams of sodium sulphate, stirring dissolved, cooled and watered down to 1000 ml, that is, made into 3,5-nitrosic acid reagent, stored in a brown bottle for backup., glucose standard curve drawingtake glucose standard liquid (1mg/ml) 0, 0.2, 0.4, 0.6, 0.8, 1.0ml in 25 ml
    test tube
    , respectively, accurately add DNS reagents 2 ml, boiling water bath heating 2min, running water cooling, water to fill up to 15ml scale. The absorbent degree is measured at a wavelength of 540nm.5, sample determinationsample fluid is appropriately diluted, so that the sugar concentration of 0.1-1.0mg/ml, the diluted sugar liquid 1.0ml in a 15 ml scale test tube, plus DNS reagent 2.0ml, boiling water boiling 2min, after cooling with water to 15 ml scale, in 540nm wave length to determine the absorption. The number of glucose mg/ml is found from the standard curve. Find out the sugar content in the sample.worried about the bio-chemical experiment report, may wish to look at this article, hope to be useful to everyone. In fact, the most important thing is to know the DNS reduced sugar content determination principle, as for the experimental reagent preparation, operation steps, these are much the same, mechanical operation. Only by mastering the principle can you understand why you should do this, and what you must pay attention to in your operation.
    .
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