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Chapter IV

Results and Discussions

This chapter presents and interprets the data gathered in textual, graphical and

tabular manner.

Results of Microbial Assay of All Treatments against S. epidermidis

(Staphylococcus epidermidis) in 24 hours

Antibacterial activity of the different concentrations of Patolang Gubat

(Trichosanthes quinquangulata) stem extract, positive and negative control were

evaluated in accordance to the Guidebook to Plant Screening; Phytochemical and

Biological, RCNS, UST. The results show that the treatments released antibacterial

activity against the standard culture strains of S. epidermidis (Staphylococcus

epidermidis) in 24 hours as shown in the table.


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Table 2: Antibacterial Screening of Patolang Gubat (Trichosanthes

quinquangulata) Stem Extract

S. epidermidis Replicates

Mean

R1 R2 R3 Zone of Descriptive
Treatments
(mm) (mm) (mm) Inhibition Interpretation

(mm)

T1 Positive Control
34 34 34 34 Strong
(Cefoxtamine)

T2 Negative Control

(Sterilized Distilled 6 6 6 6 Negative

Water)

T3 50% of Patolang
17 13 11 13.67 Moderate
Gubat Stem Extract

T4 75% of Patolang
20 17 18 18.33 Strong
Gubat Stem Extract

T5 100% of Patolang
25 23 24 24 Strong
Gubat Stem Extract

Grand Mean ZI in the 5 experimental treatments 18.67 Strong


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Figure 3: Graph of the Results of the Microbial Assay in all Treatments

40

35

30

25
Replicate 1
20
Replicate 2
15 Replicate 3

10

0
Treatment 1 Treatment 2 Treatment 3 Treatment 4 Treatment 5

The descriptive interpretation of the results was adopted from Capal et, al, which

followed the interpretative range of standard zone:

Zone of Inhibition Inhibitory Activity

> 17 +++, strong

12 -16 ++, moderate

7 - 11 +, weak

0-6 Negative

The results show that that Patolang Gubat (Trichosanthes quinquangulata) Stem

Extract strongly exhibits antibacterial activity against Staphylococcus epidermidis. T 1,

the positive control (Cefoxtamine), has the highest average mean zone of inhibition of

34mm followed by the T5 which contains 100% of Patolang Gubat Stem extract with a
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mean zone of inhibition of 24mm. this indicates that the Patolang Gubat Stem extract

has a strong antibacterial activity against Staphylococcus epidermidis.

Statistical Analysis for Screening for Antibacterial Activity of Leaves Extract

Table 3. Statistical Analysis on the Antibacterial Screening of Patolang Gubat

(Trichosanthes quinquangulata) Stem Extract against Staphylococcus

epidermidis

This table shows the computed values based on the statistical test, One Way

ANOVA, which was performed to interpret the results for the Antibacterial Screening.

Source of Variation SS Df MS F-Tabulated

Treatments 1343.06 4 335.77 3.48

Error 246.4 10 24.64 3.48

The Statistical Test indicates that the mean zone of inhibition of the different

treatments are significant. However, it could be noted that treatment 1 to treatment 4

are statistically different from treatment, which in the positive control. This means that

the four treatments are not effective against Staphylococcus epidermidis. On the other

hand Treatment 4 is not significantly different from treatment 1. This implies that 100%

of the extract has the same effectiveness or efficacy with Cefoxtamine.

Result implies that Patolang Gubat (Trichosanthes quinquangulata) has a high

potential as an antibacterial medicine.


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Results of the Phytochemical Analysis of the Patolang Gubat (Trichosanthes

quinquangulata) Stem Extract

The Patolang Gubat (Trichosanthes quinquangulata) stem extract undergone

the Phytochemical Screening in order to determine the different bioactive compounds

present on the plant. The test was done in accordance with the Guidebook to Plant

Screening: Phytochemical & Biological, RCNS, UST. The results in this test are as

follows:

Table 4: Phytochemical Screening of Patolang Gubat (Trichosanthes

quinquangulata) Stem Extract

Phytochemicals Results

Alkaloids Positive

Flavonoids Negative

Glycosides Negative

Saponins Negative

Steroids Negative

Tannins Positive

In the Phytochemical Screening, it is found out that the bioactive compounds

present in the Patolang Gubat (Trichosanthes quinquangulata) stem extract are

Alkaloids and Tannins. It is also shown, based on the results that Flavonoids,

Glycosides, Saponins and Steroids are not present in the plant material.

Tannins is an antiseptic and astringent that affected the zone of inhibition of the

plant extract against S. epidermidis.


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Results of the Thin Layer Chromatography Test (TLC) of the Patolang Gubat

(Trichosanthes quinquangulata) stem Extract

TLC was developed in six different solvents. Viewed under UV lamp and sprayed

with vanillin sulfuric acid. This test was done under concentrate condition. The results

for the Thin Layer Chromatography Test were expressed qualitatively and it is found out

that the extract is polar.

A spot was observed at Rf value0.96 with the non-polar solvent Dichloromethane

10:1. No separation of components was observed in the rest of the developing solvents

and most of the components stayed at the original spot. The components of the

PatolangGubat (Trichosanthesquinquangulata) stem may be mostly polar.

Results of the Physico-Chemical or Stability Test of Formulated Ointment

The results for the stability test done on the ointment are shown in the table. The

results include the evaluation of the ointment specifically on its colour, appearance and

consistency and pH level for a month. The ointment was observed three times for a

month.

Table 5: Stability Testing of Patolang Gubat (Trichosanthes quinquangulata) Stem

Extract

Trial Number Color Appearance pH


Uniform coloration with no liquid
1 Greenish
formation 5.0
Uniform coloration with no liquid
2 Greenish
formation 5.1
Uniform coloration with no liquid
3 Greenish
formation 5.2
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The table shows that the results the trials are stable in color and appearance.

Since there are no liquid formations, it indicates that the extract did not separate from

the base. First observation done on the ointment is 5.0 for the pH level. On the second

observation, the pH level rose to 5.1 and the third observation the pH level rose to 5.2.

This tells that the pH level of the ointment is within the acceptable range.

Figure 4: Graph of the pH level of the Formulated Ointment in All Trials

pH Level
5.25

5.2

5.15

5.1

pH Level
5.05

4.95

4.9
Trial 1 Trial 2 Trial 3

The table shows that the results for the first observation done on the ointment

are 5.0 for the pH level, it was greenish in color and uniform coloration with no liquid

formation was observed. On the second observation, the same results were gained

aside from the pH level which rose to 5.1. And on the third observation the colour of the

ointment was still greenish, uniform coloration with no liquid formation was also

observed but its pH level rose to 5.2. With respect to the scale of interpretation of pH

level, the formulated ointment is acidic.


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This indicates that the formulated ointment is stable with regards to its physical

and chemical properties for a month. This tells that the ointment is safe and there are no

changes on its chemical properties that might affect its effectiveness and other

properties when used or applied.

Results of the Antioxidant using 0.01 mM DPPH scavenging Assay of all Trials of

the Patolang Gubat (Trichosanthes quinquangulata) Stem Extract

Antioxidant Screening was done on the Patolang Gubat (Trichosanthes

quinquangulata) stem extract using 0.01mM DPPH scavenging assay. The results in

this test are shown on the table below.

Table 6: Scavenging Activity

Trial Number Level of Absorbance % Scavenging Activity

1 0.286 22.70

2 0.290 21.62

3 0.290 21.62

Graph of the Scavenging Activity of the Plant Extract in All Trials

The results for the three trials were 22.70%, 21.62% and 21.62% respectively.

The findings indicate that the Patolang Gubat (Trichosanthes quinquangulata) Stem

extract have inhibit the formation of DPPH at the percentage range of 21.62% to

22.70%.
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% Scavenging Activity
22.8

22.6

22.4

22.2

22

21.8 % Scavenging Activity

21.6

21.4

21.2

21
Trial 1 Trial 2 Trial 3

This result tells that the plant extract can be an antioxidant agent however the

effectiveness of this extract is not comparable to other commercial antioxidant

materials.