Primary Products of Photosynthesis:

Carbohydrates and Proteins in Plants

Homillano, Laki, Pascual, Perez, Sumilang
Group 4
Objectives
 To determine the presence of carbohydrates and proteins
in plants
 To be familiar with some chemical tests for plant organic
matter
 To describe some pharmacological use of these chemical
compounds
 Methodology
 Testing for Carbohydrates
Acquire 6 test tubes

Few drops of
Mashed raisin + 1mL corn syrup
sugarcane juice
3mL water + 2mL water
+ 3mL water

Test Tube A Test Tube B Test Tube C

Pinch of banana Scraped potato 3mL distilled

+ 2mL water + 2mL water water

Test Tube D Test Tube E Test Tube F

Shake test tubes until materials are mixed well

Methodology
 Fehling’s Test

Add 3mL of Fehling’s solution in each test tube

Boil test tubes in a water bath

Positive Result:
Brick red precipitate
indicates presence of sugars
Methodology
 Benedict’s Test

Add Benedict’s solution in each test tube

Heat test tubes in a water bath

Positive Result:
Brick red precipitate
indicates presence of reducing sugars
Methodology
 IKI reagent test

Put specimen in microscope slides

Add IKI solution and observe under the

microscope

Positive Result:
Brown IKI turns to blue-black color
Methodology
 Testing for Proteins (Xanthoprotein Test)

Soak mongo & corn seeds overnight

Ground using mortar and pestle

Add water and let it stand for

30 minutes

Filter the extracts

Methodology
 Testing for Proteins (Xanthoprotein Test)
Put 3mL of mongo filtrate, corn
filtrate, and soya milk in separate
test tubes

Add 1mL of concentrated nitric acid

Boil in water bath for 5 minutes

White precipitate forms, turns yellow

when heated
Methodology
 Testing for Proteins (Xanthoprotein Test)

Cool the solution

Add 8-10 mL of 10% NaOH

Yellow color becomes orange

Results

A. Carbohydrates
1. Fehling’s Solution

Group 1 Group 2 Group 3 Group 4

Sugar solution Positive Positive Positive Positive
Raisin Positive Positive Positive Positive
Corn Syrup -/- -/- -/- -/-
Banana Negative Positive Positive Positive
Potato Negative Negative Negative Negative
Water Negative Negative Negative Negative
Results

A. Carbohydrates
1. Fehling’s Solution

Banana Raisin Sugar Solution Potato Water

Results

A. Carbohydrates
2. Benedict’s Test
Group 1 Group 2 Group 3 Group 4
Sugar solution Positive Positive Positive Positive
Raisin Positive Positive Positive Positive
Corn Syrup -/- -/- -/- -/-
Banana Negative Negative Negative Negative
Potato Negative Negative Negative Negative
Water Negative Negative Negative Negative
Results

A. Carbohydrates
2. Benedict’s Test

(Left to Right) Banana, Raisin, Sugar Solution, Potato, Water

Results

A. Carbohydrates
3. Iodine Test
Group 1 Group 2 Group 3 Group 4
Raisin Negative Negative Negative Negative
Banana Positive Positive Positive Positive
Potato Positive Positive Positive Positive
 Results

A. Carbohydrates
3. Iodine Test

Raisin Banana Potato

Results

B. Proteins
1. Xanthoprotein Test Reagents
Group 1 Group 2 Group 3 Group 4
Mongo Nitric acid and Nitric acid and Nitric acid and Nitric acid and
10% NaOH 10% NaOH 10% NaOH 10% NaOH
Soya Nitric acid and Nitric acid and Nitric acid and Nitric acid and
Milk 10% NaOH 10% NaOH 10% NaOH 10% NaOH
Corn Nitric acid and Nitric acid and Nitric acid and Nitric acid and
10% NaOH 10% NaOH 10% NaOH 10% NaOH

2. Results
Group 1 Group 2 Group 3 Group 4
Mongo Orange Orange Orange Orange
Soya Milk Deep Orange Deep Orange Deep Orange Deep Orange
Corn Yellow Yellow Yellow Yellow
Results

B. Proteins
3. Iodine Test

Corn Seeds Soya Milk Mongo Seeds

Discussion
A. Carbohydrates
1. Fehling’s Test
Positive Result: Presence of brick red precipitate
The Fehling’s solution is a mixture of two solutions:
Fehling’s Solution A: copper sulfate (CuSO4) solution
Fehling’s Solution B: a strong base (ex. KOH)
When mixed, these two react to produce cupric hydroxide (Cu(OH)2).
When an alkaline solution of cupric hydroxide is heated in the
presence of a reducing sugar, it gets reduced to a yellow- or red-colored
precipitate which is cuprous oxide (CuO).
 Reducing sugars respond positively to this test. A reducing sugar has an
aldehyde or ketone group that causes the reduction of cupric
hydroxide to cuprous oxide. Examples of these sugars include glucose,
galactose, fructose, lactose, and maltose.
Discussion
A. Carbohydrates
2. Benedict’s Test
Positive Result: Presence of brick red precipitate
Similar to Fehling’s test, but more sensitive. Benedict’s solution
composed of just one solution, which is a copper sulfate solution with a
weak acid such as sodium carbonate or sodium citrate as buffer.
Reducing sugars are detected because of the reduction of copper (II)
ions, found in copper sulfate, to copper (I). Cuprous oxide, which has
copper (I) ions, is the red-colored precipitate formed by this reaction.
 It is considered to be more reliable compared to the Fehling’s Test
because the strong alkalinity of the Fehling’s solution may cause the
breakdown of sugars which can lead to different results. Also, the
Benedict’s test produces a large amount of precipitate, so it can detect
even low levels of reducing sugar/s.
Discussion

A. Carbohydrates
3. Iodine Test
Positive Results: Brown IKI turns to Blue-Black color
Iodine, in an aqueous solution of potassium iodide, reacts
with starch producing the blue-black coloration
A complex of iodine forms, which then is able to slip through
the helix of the amylose (polysaccharides trap the iodine)
Discussion
A. Carbohydrates
1. Fehling’s Test and Benedict’s Test
Interpretation of Results:
Sucrose solution: sucrose is not a reducing sugar, so the results should be
negative. However, since the solution was heated, it may
have broken down to glucose and fructose, which respond
positively to the tests above.
Raisin: All positive because a fruit has fructose
Banana and Potato: these contain starch, which is a polysaccharide. They
should produce a negative result, unless the
polysaccharide/s have been broken down to a
reducing sugar
Water: negative control
Discussion

2. Iodine Test
Raisin: negative, for it doesn’t have starch. May be misinterpreted
due to its originally dark color.
Banana and Potato: positive, with blue-black-colored starch grains after
reacting with the originally brown IKI solution
Discussion

B. Proteins
1. Xanthoprotein Test
Positive Results: Xanthoprotein presence is indicated by yellow
coloration. Adding NaOH turns the yellow color into orange.
Aromatic Rings are present in the filtrate and these react with the nitric
acid that is added to the solution, thus creating the yellow color.
Xanthoproteic acid is formed. NaOH is added to neutralize the acidic
filtrate resulting to the change of color from yellow to orange. It is
also used to verify the presence of proteins. The darker the orange
color is, the greater the amount of protein present.
Discussion

B. Proteins
1. Xanthoprotein Test
Interpretation of Results:
Soya Milk: very rich in protein
Mongo Beans: also has protein but less compared to the soya milk
Corn Seeds: least amount of proteins, probably enzymes present in the seed
Guide Questions
Medicine/Drug Main Ingredient Test Reagent Function of Drug
Present
Dextrose Glucose Fehling’s / Hypoglycemia
Benedict’s Test management
Laxatives Glycerin/Fatty Grease Spot Test Increase in bowel
Acid movement
Calamine Lotion Zinc oxide Xanthoprotein Test Anti-irritation
Caramel Sucrose Fehling’s / Drug flavoring
Benedict’s Test
Castor Oil Fatty Acid Grease Spot Test Treats constipation
Papain Amino Acid Xanthoprotein Test Anti-inflammation
Trypsin Amino Acid Xanthoprotein Test Osteoarthritis
treatment
Mint Fatty Acid Grease Spot Test Treatment for
stomach ache
Guide Questions

Why have plants been used as sources of many pharmaceutical products? How is
this related to photosynthesis
 Plants contain various substances with pharmaceutical properties that are found
because they are mostly produced via photosynthesis, a process that only plants can
do.

Give the importance of using chemical tests in determining plant constituents.

 Plants don’t all have the same constituents. They also produce a lot of different
substances. The main purpose of testing it is to find out where we can get things that
we need specifically.
References

 Nigam, A. & Ayyagari, A. (2007). Lab manual in biotechnology, immunology

and biotechnology. Tata McGraw-Hill Publishing Company Limited: New Delhi.
 http://chemwiki.ucdavis.edu/Biological_Chemistry/Carbohydrates/Case_Stud
ies/Starch_and_Iodine
 http://chemwiki.ucdavis.edu/?title=Organic_Chemistry/Aldehydes_and_Keto
nes/Reactivity_of_Aldehydes_%26_Ketones/Oxidation_of_Aldehydes_and_Keto
nes
 http://www.bbc.co.uk/bitesize/standard/chemistry/plasticsandothermateria
ls/carbohydrates/revision/3/
 http://brilliantbiologystudent.weebly.com/benedicts-test-for-reducing-
sugars.html