PROCEDURE

A. Food Reserves Test

1. The whole of the cotyledon was stained in iodine solution.
2. Results are recorded in Table 2.
3. Blender used to prepare the sample for reducing sugar test.
4. The test tubes are filled with 1 g sample.
5. 4 ml Benedict’s reagent added into the test tube and the initial color recorded.
6. The test tube was placed in a boiling water bath for 3 minutes.
7. The test tube then removed form the hot water bath using the test tube holder and
placed in test tube rack. The color changes recorded in Table 2.
8. The contents of the test tube were discards and the test tube was rinsed properly.
The excess water was shaken out from the test tube.
9. The same procedure was carried for other samples.

B. Effect of Amylase on a Starchy Substance

1. 10 ml of uncontaminated saliva was collected in a measuring cylinder: the flow of
saliva was 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. Another 2 ml was transferred to another test tube and 2 drops of hydrochloric acid (1
mol dm-3) was added, the solution was mixed well and left for 15 minutes. The rest of
untreated saliva were kept at the room temperature.
4. Six pairs of test tubes were set up as follows, each test tube was labeled with code
number as in Table 1.
Table 1: Test Tube Setup
LABEL CONTENT
1 2 ml sugar only
PAIR A 2 2 ml sugar only
3 2 ml sugar only
PAIR B 4 2 ml sugar only
5 2 ml untreated saliva only
PAIR C 6 2 ml untreated saliva only
7 2 ml starch + 1 ml untreated saliva
PAIR D 8 2 ml starch + 1 ml untreated saliva
9 2 ml starch + 1 ml preheated saliva
PAIR E 10 2 ml starch + 1 ml preheated saliva
11 2 ml starch + 1 ml acidified saliva
PAIR F 12 2 ml starch + 1 ml acidified saliva
5. The test tube was left for 10 minutes before the next procedure.
6. The contents of each pair of test tube was tested with:
a) For starch, 2 drops of Iodine solutions were added.
b) For sugar, heated with 3 ml of Benedict’s reagent.
7. Starch test were done on the first of each pair of test tube, and sugar test are done
on the second test tube of each pair.
8. The test tube was arranged in the rack in numerical order to facilitate comparison.
9. Color changes are recorded in Table 3. For each starch and sugar contents, the test
tube which shows either positive or negative results was indicated.
RESULTS
A. Food Reserves Test

Table 2: Analysis of Seed Characteristic

Food Reserve Test
(present or not present)
Sample Starch Reducing Sugar

Peanut Not present Not present

Mung Bean Not present Not present

Corn Present Present

Figure 1: Iodine Test on Food Reserves

Figure 2: Benedict’s Test on Food Reserves

B. Effect of Amylase on a Starchy Substance

 Table 3: Enzyme Reactions for Sugar and Starch
TEST TUBE
LABEL CONTENT OBSERVATION CONCLUSION
PAIR 1 2 ml sugar only No change No presence of
A
starch
2 2 ml sugar only Color change from Presence of
blue to red-brick sugar
precipitate
PAIR 3 2 ml sugar only Yellow to blue-black Presence of
B
starch
4 2 ml sugar only No change No presence of
sugar
PAIR 5 2 ml untreated saliva only No change No presence of
C
starch
6 2 ml untreated saliva only No change No presence of
sugar
PAIR 7 2 ml starch + 1 ml untreated Yellow to light blue Presence of
D
saliva starch
8 2 ml starch + 1 ml untreated Blue to greenish Presence of
saliva yellow sugar
PAIR 9 2 ml starch + 1 ml preheated Yellow to blue Presence of
E
saliva starch
10 2 ml starch + 1 ml preheated No change No presence of
saliva sugar
PAIR 11 2 ml starch + 1 ml acidified Yellow to blue-black Presence of
F
saliva starch
12 ml starch + 1 ml acidified No change No presence of
saliva sugar
Figure 3: Effect of Amylase on a Starchy Substance

Figure 4: Benedict’s Test on a Sugar Substance