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Dextrose Estimation via Colorimetry

The document outlines an experiment aimed at estimating dextrose concentration using colorimetry, which measures the absorption of light at specific wavelengths based on Beer-Lambert’s law. It details the necessary materials, preparation of solutions, and the procedure for measuring absorbance with a colorimeter to determine the concentration of dextrose in a sample. The conclusion emphasizes that the reaction between dextrose and p-hydroxybenzoic acid hydrazide in an alkaline medium allows for quantification of dextrose through colorimetric analysis.

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49 views2 pages

Dextrose Estimation via Colorimetry

The document outlines an experiment aimed at estimating dextrose concentration using colorimetry, which measures the absorption of light at specific wavelengths based on Beer-Lambert’s law. It details the necessary materials, preparation of solutions, and the procedure for measuring absorbance with a colorimeter to determine the concentration of dextrose in a sample. The conclusion emphasizes that the reaction between dextrose and p-hydroxybenzoic acid hydrazide in an alkaline medium allows for quantification of dextrose through colorimetric analysis.

Uploaded by

drypagrawal21
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd

Estimation of dextrose by Colorimetry

AIM- Aim of the experiment is to estimate sucrose by colorimeter


BACKGROUND Colorimetry is the measurement of concentration of analyte in a
solution by determining absorption of a particular wavelength. The technique is based
upon Beer-Lambert’s law. Overall the process is accomplished with the measurement of
analyte in a colored solution or development of color that appears in the solution
following reaction with specific reagent. In alkaline medium p-hydroxybenzoic acid
hydrazide (PHBAH) reacts with dextrose that give products which can be assayed by
colorimetry. Dextrose reacts with p-hydroxybenzoic acid hydrazide (PHBAH) reagent to
give coloured product; the absorbance noted is proportional to concentration of sugar.
REQUIREMENTS
Dextrose: 5mg, Sodium hydroxide: 3-5 pellets, p- hydroxybenzoic acid hydrazide
(PHBAH): 50 ml; Test tubes, Burette, Colorimeter, Water bath
PROCEDURE
Alkaline solution:- Put 3-5 pellets of sodium hydroxide in about 50 ml of water, stir to
mix well.
Diluent:- Add 10.0 ml of p-hydroxybenzoic acid hydrazide (PHBAH) solution with
30.0ml of water along with few drops of Sodium hydroxide.
Standard stock solution preparation
Weigh 5.0 mg of dextrose; add in a test tube containing 15.0 ml of water, 2-3 drops of
sodium hydroxide and 5.0 ml of p-hydroxybenzoic acid hydrazide (PHBAH) solution,
and mix well.
Working standard solution preparation Dilute 5.0ml of stock solution with 15.0 ml of
diluent.
Sample solution preparation Take a test tube, add 1.0 ml of sample with burette, add
4.0 ml of water to it, and shake gently to mix. Keep the above sample solution for 30 -35
minutes at normal temperature. Again shake gently, then add 5.0 ml of p-
hydroxybenzoic acid hydrazide (PHBAH) coloured solution and few drops of sodium
hydroxide, mix well and keep the test tube in water bath in boiling mode. After about 5
minutes withdraw test tube from water bath, allow to cool, then add 10 ml of water and
mix well. Instrumentation Before measuring absorbance by colorimeter clean the glass
tube with water, and ensure zero absorbance with diluent. To finalize the light source,
take sample solution in glass tube and keep first in blue light source and repeat with the
available light source simultaneously recording absorbance. Use the light source with
highest absorbance and record the absorbance for the followings. 1. Standard stock 2.
Working standard solution 3. Sample of unknown concentration.
Note the above three different absorbance. Now plot the graph of two known solution
(1 is standard stock, 2 is working standard) i.e. absorbance (Y-axis) against
concentration (X-axis), plot a straight line in between two points and find the
concentration of unknown sample with the sample absorbance. CONCLUSIONThe
chelate formed from the reaction of sucrose and p-hydroxybenzoic acid hydrazide
(PHBAH) in alkaline medium is measured by colorimeter which is the basis of
quantifying the amount of sucrose present in sample.
REFERENCES
1. [Link] Accessed on 5
August 2017.
2. Lever M. Colorimetric and Fluorometric Carbohydrate Determination with
pHydroxybenzoic Acid Hydrazide. Biochem Med. 1973;7: 274-81

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