FACULTY OF SUSTAINABLE

AGRICULTURE

RT10303

CHEMISTRY FOR AGRICULTURE

DR. LUM MOK SAM

PRACTICAL TASK 4

(17/11/2020)

DETERMINATION OF TOTAL CARBOHYDRATE IN PLANT SAMPLES (PHENOL-

SULPHURIC ACID METHOD)

Prepared by:

NOOR AMYRAH ALINA BT. MOHD AMERUL ‘ASRI BR20110034

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TITLE:

Determination of Total Carbohydrate in Plant Samples using Phenol-Sulphuric Acid Method.

OBJECTIVE:

To determine the total carbohydrate in plant samples using phenol-sulphuric acid method

INTRODUCTION:

Carbohydrates (also referred to as saccharides) are molecular compounds composed of

only three components: carbon, oxygen and hydrogen. The comparatively small molecules
are monosaccharides (e.g. glucose) and disaccharides (e.g. sucrose). Sometimes they are
called sugars. Other carbohydrate molecules (polysaccharides like starch and cellulose) are
rather large.
The following are carbohydrates:

 Source of energy for the body e.g. glucose and a store of energy, – for example starch
in plants
 Building blocks for polysaccharides, such as cellulose in plants and glycogen in the
human body (giant carbohydrates)
 Other molecular elements, such as DNA, RNA, glycolipids, glycoproteins, ATP,

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APPARATUS:

1. 8 Test Tube
2. Pipette
3. Volumetric flask
4. Distilled water
5. Spectrophotometer Machine

Reagents
1. Phenol 5% solution:
 Carefully weighed the pure phenol and dissolved in 100 ml distilled water
2. Sulphuric Acid Concentrated (98%)
3. Standard Glucose Solution:
 100mg of dry glucose powder was dissolved in 100ml of distilled water
 10 ml of this solution was diluted to 100ml with distilled water (100 micrograms/ml)

PROCEDURE:

Firstly, aliquots of standard solutions 0, 0.2, 0.4, 0.6, 0.8, and 1.0 ml was pipette out into 6
test tubes. Then, 0.5 ml of sample extract was pipette out into other two test tubes. 1.0,
0.8, 0.6, 0.4, 0.2, 0.0 ml and 0.8 ml distilled water was added in respective test tubes to
make final volume to 1.0 ml in each tube. Next, 1ml phenol was added to each tube
followed by 5ml concentrated H2SO4 solution in each tube and mixed well. Lastly, read the
absorbance after 10 minutes of pink colour after setting zero with 1 st test tube(blank). The
graph was plotted with the readings and concentration of glucose in aliquots of standard
solution.

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RESULTS:

Reagent Tube 1 Tube 2 Tube 3 Tube 4 Tube 5 Tube 6

Gluocose (mL) 0.0 0.2 0.4 0.6 0.8 1.0
Gluocose (Ug) 0 20 40 60 80 100
Absorbance 0.0 0.12 0.21 0.36 0.47 0.59
(490nm)

Calculation:

a) The glucose content

Tube 2

b) The standard curve

c)

DISCUSSION

In this experiment, it shows that a weak acid is an acid that partially ionizes or dissociates in
water to create a low hydrogen ion concentration. It indicates that the percent
concentration of acetic acid in vinegar is 0.35% after the measurement, while citric acid is
0.68% in fruit juice. Therefore, citric acid in fruit juice has the stronger percent strength
with 0.68% than the acetic acid in vinegar with only 0.35%. Thus it shows that the pH of an
acid solution changes based on the concentration of the acid affects the concentration of
hydrogen produced.

CONCLUSION

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In conclusion, a titration used to display the equivalence point to determine the
concentration of acetic acid in vinegar and citric acid in fruit juice. Therefore, it was crucial
to add the correct amounts of deionized water and NaOH to both vinegar and fruit juice,
and also extremely important to take and record the correct measurements. The
concentration was found to be 0.68% citric acid in fruit juice which is stronger than acetic
acid in vinegar with only 0.35%.

REFERENCES

 Fraser, J.R., & Holmes, D.C.,J. Sci. Fd. Agric., 1956, 7, 589,
 Chang,R 2005, Physical Chemistry for the Biosciences. USA: University Science Books.
 Spechphotometry 1995, Nature; London. Vol. 374, no. 6521, p. 481, Nature Publishing
Group, London