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ferrin "> reagents color method
" (purposes and requirements)
1, mastering the principles and methods of quantitative determination of reduced sugar
2, learncharcodometer use
.3, familiar with the production method of standard curves
< p style-text-align:left;">"Ferrin reagent color method is the use of Filin reagents After mixing A and B, the sodium sulfate of A liquid and sodium hydroxide of a liquid produce sky-blue copper hydroxide precipitation, which reacts with potassium saline of acetylene in alkaline solution to produce complexcompound (potassium saloonate sodium copper).
which has a slightly different sky blue color from ordinary copper salts, but still has all the normal reactions of copper salts. The sky-blue potassium sodium sodium sodium copper has the largest absorption peak at a wavelength of 590nm.
monosaccharides and polysaccharide hydrolytes contain reductive aldehydes or ketones, boiling under alkaline conditions can reduce the two-priced copper ions in the Filin reagent to a priceless copper oxide, and the blue Filin reagent decolorization, the degree of decolorization is directly related to the sugar content in the solution.
This is where the light absorption value of the solution is measured at a wavelength of 590nm and a standard curve is made to find the sugar content in the sample.
the advantage of this method is that it is easy to operate, Filin reagents can not be rationed, the measurement error is ±0.02%, but must be regularly corrected by the standard curve. The disadvantage is that the reagent is unstable, needs to be made temporarily, and is easily disturbed by muscle liver (acid) and so on, which is very disadvantageous in medical analysis. Its measurement range is 0.1 to 0.5 mg/mL reduced sugar, if the sugar content exceeds this range, the error is significantly increased.
"
1,""standard curve production
pressed table to add 1 mg/> mL glucose standard solution 0,0.5, 1.0, 1.5, 1.5, respectively, in each2.0, 2.5, 3mL, plus distilled water to 3mL, 0,0.5, 1.0, 1.5, 2.0, 2.5, 2.5, each tube containing glucose 3.0mg, respectively, in each tube to add 2mL filin reagents, mixed, in the test tube port with a glass ball cover, put into the boiling water bath heating15min, removed and cooled in running water, After 1500r/min centrifugation 15min, the liquid was taken at 590nm wavelength for color determination, with distilled water as a color ratio, reading the light density value of each tube containing different concentrations of sugar and the light density value of the blank tube. Then the light density value of the blank tube minus the light density value of the different concentrations of sugar in each tube, and use this value as a ordinate, the relative sugar content as a horizontal coordinate, to draw the standard curve.
reagent tube number | 0 | 1 | < left;">2 |
3
4
5
d reduced sugar
< td width s"45" >left; "> total sugar
0
0.5
1.0
1.5
2.0
2.5
3.0
1.5
1.5
ferrin reagents/mL
2
2
2
-left;">2
2
2
2
2
2
water/mL
3
. 5
2
1.5
0.5
0
<: left; >1.5
1.5
total volume/mL
5
5
-align:left;">5
5
5
5
5
5
5
<: left;" > A590