Jan 04, 2017
How To Test Polysaccharide?
1.2 Redistilled Water
1.4 5% Phenol Solution: Take heat phenol distillation and distillate collect182 degree, absorb distillation10ml and add water to200ml.
2. Instrument and Appliance
2.1 Electronic Balance
2.2 Glass Apparatus: Volumetric Flask
2.3 Ultrasonic Cleaner
2.4 Ultraviolet Spectrophotometer
2.5 Glass Cell
3.1 Established Standard Curves
3.1.1 Accurately measure constant weight Glucose reference substance10mg and add into100ml volumetric flask with heavy steam water dissolved and constant volume.
3.1.2 Accurately absorb above standard liquid 0, 0.1, 0.2, 0.4, 0.6, 0.8ml, respectively in Graduated test tube with stopper, add distilled water to2ml, and add1.0ml5% Phenol Solution， Sulfuric Acid5.0ml, Vibration wave5 minutes, make a blank control at the same time.
3.1.3 Heating for15 minutes in a boiling water, take out and cooling in cold water for30 minutes.
3.1.4 Uv-vis spectrophotometer absorbance at490 nm wavelength.
3. 1 .5 Absorbance as ordinate, glucose concentration as abscissa, drawing standard curve.
3.2 Sample Test
3.2. 1 Accurately measure sample 0.2g into soxhlet extractor, with10 ml80% ethanol reflux2 h, and Residue with80% ethanol to wash to colorless, then with100ml water reflux2h,
repeated washing into250 ml volumetric flask, constant volume, Shake well to the sample liquid.
3.2.2 Accurately absorb above standard liquid 2.0ml, add into Graduated test tube with stopper, According to the standard curve"add 5% Phenol Solution",
3.2.3 Find out the sample number of mg absorbance values
V-- Dilution ratio;
A-- Sample absorbance corresponding mg