FAKULTI SAINS GUNAAN

UNIVERSITI TEKNOLOGI MARA

CAWANGAN SARAWAK KAMPUS SAMARAHAN 2

BIO270 – ANIMAL AND PLANT PHYSIOLOGY

SCIENTIFIC LAB REPORT

NAME: STUDENT ID:

NUR HAZIQAH MAULAD MOHAMAD NADZAR 2022848842

ABDILLAH ADIB BIN ADAM 2022826212

ABANG MOHAMAD ADIL BIN ABANG MOHAMAD 2022493412

BUSRAH

MUHD IRFAN HAKIMI BIN OMAR ALI 2022857368

GROUP: AS1203C

EXPERIMENT TITLE: 3
DETERMINATION OF STARCH AND SUGAR IN SEEDS AND HOW THOSE
STARCHES BEEN DIGESTED IN HUMAN

LECTURER: DR SYAZANI BINTI DARHAM

DATE OF SUBMISSION: 15TH JANUARY 2024

INTRODUCTION:
Starch is polysaccharide which must be broken down into free sugar molecules
before absorption can take place. Starch is mixed with saliva under various conditions and
after a given period the mixture is tested for starch with iodine solution and for sugar with
Benedict’s reagent.

The aim of our team in the experiment examining the function of amylase in food
digestion is to understand how starch breaks down in lowering sugar form from
polysaccharide to monosaccharide. Saliva contains an enzyme called amylase, which is
essential to this process. Amylase catalyses the breakdown of starch into the disaccharide
maltose and the monosaccharide glucose when starch and saliva are. The body needs this
transformation to absorb carbohydrates. Benedict's reagent is used to test the mixture for
the presence of reducing sugars and the presence of starch using an iodine solution. The
results of the experiment will bring light on how amylase functions in the early phases of
digestion, specifically in the conversion of complex carbohydrates into more easily absorbed
forms.

OBJECTIVES:
1. To identify the presence of starch and reducing sugars in seeds.
2. To inspect the hypothesis that saliva contains an enzyme which break down starch to
sugar.
3. To investigate the properties of the enzyme and the conditions under which it works
most efficiently.

HYPOTHESIS:
Saliva contains an enzyme, amylase which break down starch to sugar.

MATERIALS:
I. DETERMINATION OF STARCH AND SUGAR IN PLANT SEEDS
1. Balancing scale
2. Forceps
3. Test tube
4. Test tube rack
5. Water bath
6. Dish
7. Razor blade
8. Benedict’s Reagent (Food reserve test)
9. Iodine solution (Food reserve test)
10. 0.5% tetrazolium solution (Metabolic activity test)
11. Distilled water
12. Broad bean seed (dry) and broad bean seed soaked for 24 and 48 hours (dicotyledon)
13. Corn grain (dry) and corn grain soaked for 24 and 48 hours (monocotyledon)

II. DIGESTION OF STARCH IN MAMMALS

1. Test tube (12/group)
2. Test tube rack
3. Measuring cylinder (20 cm³)
4. Water bath
5. Bunsen burner with tripod and gauze
6. Wax pencil
7. Starch suspension
8. Dextrose (glucose)
9. Iodine solutions
10. Benedict's reagent
11. Hydrochloric acid (1.mol dm³)
12. Paraffin wax

PROCEDURE:
A. Seeds Anatomy test
1. The whole of the cotyledon was stained in iodine solution.
2. Result was recorded in Table 2.
3. A blender was used to prepare the sample for reducing sugar test.
4. The test tube was filled with 1g sample.
5. 4 ml Benedict’s reagent was added into test tube and the initial colour was recorded.
6. The test tube was placed in a boiling water bath for 3 minutes.
7. The test from hot water bath was removed with the test tube holder and the test tube
was placed in rack. The colour changes were recorded in Table 2.
8. The contents of the test tube were discarded and rinsed properly. Excess water from
the test tube was shaken out.
9. The same procedure was carried out for others sample.

B. Effects of Amylase on a Starchy Substance

1. 10 ml of uncontaminated saliva was collected in a measuring cylinder. The flow can
be increased by chewing paraffin wax.
 2. 2 ml of the saliva was transferred to a test tube and placed in a boiling water bath for
15 minutes.
3. A further 2 ml was transferred to another test tube and added with 2 drops of
hydrochloric acid (1 mol/dm3), mixed well and left for at least 15 minutes. Kept the
rest of the untreated saliva at room temperature.
4. Six pairs of test tubes were set up as follows, each test tube was labelled with code
number.
5. The six pairs of the tubes been set up as follows, and each tubes labelled with code
number which were ; for iodine test ; 4mL sugar (I1), 4mL starch (I2), 4mL untreated
saliva (I3), 4mL starch + 2mL untreated saliva (I4), 4mL starch + 2mL preheat saliva
(I5), and 4mL starch + 2mL acidified saliva (I6) while for benedict test; 4mL sugar
(B1), 4mL starch (B2), 4mL untreated saliva (B3), 4mL starch + 2mL untreated saliva
(B4), 4mL starch + 2mL preheat saliva (B5) and 4mL starch + 2mL acidified saliva
(B6).
6. The test tubes had been leaving for 10 minutes before proceeding further.
7. The contents of each pair of test tube had been tested which for starch, two drops of
iodine solutions added while for sugar, with one-eight test tube of Benedict’s reagent
heated.
8. The starch test was done on the first of each pair of test tube and the sugar test was
done on the second of each pair.
9. The test tubes were arranged in the rack in numerical order to facilitate comparison.
10. The colour changes recorded. Which the test tubes show positive results and which
ones show negative results for each starch and sugar contents will be indicated.

\
DATA:

Image 1: Before iodine and Benedict’s solutions

Image 2: After Iodine Test

 Image 3: After Benedict’s Test

Image 4: Starch (B-bean and C-corn)

RESULTS:
TABLE 1: ANALYSIS OF SEED CHARACTERISTICS
Sample Food Reserve Test

(present or not present)

Starch Reducing Sugar

Peanut Not present Present

Mung bean Not present Present

Corn Present Not present

TABLE 2: ENZYME REACTIONS FOR SUGAR AND STARCH

TEST TUBE CONTENT OBSERVATION CONCLUSION
LEVEL

PAIR A 1 4mL sugar only No change No presence of

starch

2 4mL sugar only Blue to brick red Presence of starch

PAIR B 3 4mL sugar only Yellow to blue black Presence of starch

4 4mL sugar only No change No presence of

starch

PAIR C 5 4mL untreated No change No presence of

saliva only starch

6 4mL untreated No change No presence of

saliva only starch

PAIR D 7 4mL starch plus Yellow to light blue Presence of starch

2mL untreated
saliva only

8 4mL starch plus Blue to yellow green Presence of starch

2mL untreated
 saliva only

PAIR E 9 4mL starch plus Yellow to blue Presence of starch

2mL pre-heated
saliva only

10 4mL starch plus No change No presence of

starch
2mL pre-heated
saliva only

PAIR F 11 4mL starch plus Yellow to blue black Presence of starch

2mL acidified saliva

12 4mL starch plus No change No presence of

starch
2mL acidified saliva
DISCUSSION:
The endosperm, which serves as the main food storage tissue in seeds and is rich in
starch, has more stored starch than the cotyledon. It is also essential for providing nutrition
to the developing embryo in seedlings. Notably, Table 2 looked at the endosperm of maize
rather than the cotyledons of peanuts and mung beans. When iodine was applied, the brown
colour turned to a deep blue, showing the presence of starch in maize, thereby producing a
good result. The goal of the food reserve test is to determine whether the given seeds
contain reducing sugars and starch.

To enable the occurrence of interactions between enzymes and starch in the study of
starch and sugar digestion in animals, test tubes were left undisturbed for at least 10
minutes (Testing for Starch, Sugar, and Protein in Foods) The tests in test tubes 1 through 6
are used to determine whether the samples contain sugar and starch. Notably, test tube D8,
designated as such, showed a blue to greenish colour shift. This change suggested the
presence of the amylase enzyme, which efficiently breaks down starch into separate units of
glucose. Benedict's test is used to identify glucose, hence the greenish colour in D8
indicated the presence of sugar from amylase-facilitated starch-to-glucose breakdown. It is
important to notice that because the saliva had been heated beforehand, test tube E10 did
not react with Benedict's solution even in the presence of saliva. Salivary amylase became
denaturized due to the high temperature. Likewise, salivary amylase denatured in an acidic
environment, which is why test tube F12 failed the Benedict's test. Saliva acts as a digestive
enzyme on starch, breaking it down into smaller carbohydrate molecules like maltose.
Salivary amylase is released by the salivary glands in the mouth cavity. An increased
concentration of maltose in the mouth may contribute to the heightened sweetness of food.

In conclusion, the positive result in test tube D8 supported the theory that saliva,
which contains amylase, converts starch to sugar. On the other hand, test tubes 10 and 12,
which contained saliva that had been heated and acidified, respectively, did not exhibit any
sugar. This indicates that high temperatures and acidic environments can cause salivary
amylase to become denaturized, which results in the absence of sugar in the Benedict's test.

CONCLUSION:
Thus, we can infer that saliva contains the amylase enzyme, which is responsible for turning
starch into sugar. Temperature and pH have an impact on proteins referred to as enzymes;
research suggests that 37.5 °C and pH 7 are the ideal values for these proteins to operate.
The theory is approved.