CHAPTER 4

Presentation, Analysis and Interpretation of Data

This chapter includes the findings, analysis and interpretation of data obtained in

the result of the determination of Antibacterial activity of the semi-purified Flavonoid

extract of santan (Ixora chinensis lam. Family rubiaceae) leaves and the formulated

antibacterial liquid hand soap against pathogens.

1. Extraction and identification of Semi-Purified Flavonoids

The percentage yield was computed with the data of weight of the sample and

weight of the residue.

Table 1

Percentage Yield

Weight of the sample 200g

Weight of the residue 12.73g

% Yield 6.37%

Table 1 shows that in a sample weighing 200g and the semi-purified flavonoid

extract weighing 12.73g gave a result of 6.37% percentage yield.

2. Physical Test for flavonoids

Table 2

Results of Organoleptic Test and Solubility Test

Test Result

Organoleptic

Color Light brown

Odor Pungent

Appearance Powder form

Solubility

Distilled water Soluble

80% Ethanol Soluble

Acetone soluble

Chloroform Insoluble

Ether Insoluble

Table 2 shows the organoleptic test was conducted to ascertain the odor, color,

and the consistency of the semi-purified flavonoid extract. For the solubility test, the

flavonoid was soluble with Distilled water, 80% Ethanol and Acetone.
2.2 Chemical Test for flavonoid

Table 3

Results of the Chemical Test of Semi-purified Flavonoid Extract

Test
Expected Result Actual Result Interpretation
performed

Shinoda’s Test Yellow coloration Yellow Presence of flavonoid

Intense yellow or
AlkalineTest Yellow Absence of flavonoid
colorless

Lead Acetate Formation of Formation of

Presence of flavonoid
Test precipitate precipitate

Table 3 shows that the flavonoids was found to be present in Santan (Ixora

chinensis) leaves.
2.3 FTIR (Fourier Transform Infrared) Analysis

The FTIR was used to discern the functional groups present in Santan (Ixora

chinensis) leaves that would be the basis for the identification of the constituent present.

Table 4

Results of FTIR Analysis

Absorption Intensity Functional group Peak

(cm -1)

Medium 44.59 Aldehyde 2855.14

Weak, Broad 47.88 Alcohol 3235.33

Strong 32.87 Carbonyl 1729.48

Table 4 shows that there is a quercetin present in the extract base from the

standard
3. Microbiological Test of the Flavonoid Extract

Table 5

Results of Zone of Inhibition by the Treatment of Semi-Purified Flavonoid

Extract on Escherichia coli

Result Zone of Interpretation

Inhibition in
(mm) (mm)
Negative Control Mean 0
No. of Trial 0 0 Inactive
0
Positive Control Mean 27.70
No. of Trial 25.40 25.42 Very Active
23.15
Flavonoid extract Mean 8.30
(250mg/ml) No. of Trial 9.90 8.43 Inactive
7.10
Flavonoid extract Mean 11.50
(500mg/ml) No. of Trial 11.70 10.3 Partially
Active
7.70
Flavonoid extract Mean 11.90
(1000mg/ml) No. of Trial 20.30 13.57 Partially
Active
8.50

Table 5 shows that the negative control is inactive and positive control and the

flavonoid extract 250mg/ml is inactive and 500mg/ml, 1000mg/ml is partially active.

 Table 6

Results of Zone of Inhibition by the Treatment of Semi-Purified Flavonoid Extract

on Staphylococcus aureus

Result Zone of Interpretation

Inhibition in
(mm)
(mm)

Negative Control Mean 0

0 0
No. of Trial Inactive
0
Positive Control Mean 25.25
26.10 24.55
No. of Trial Very Active
22.30
Flavonoid extract Mean 24.20
23.10 24.07
(250mg/ml) No. of Trial Very Active
24.90
Flavonoid extract Mean 24.70
24.40 24.73
(500mg/ml) No. of Trial Very Active
25.10
Flavonoid extract Mean 24.15
23.30 23.95
(1000mg/ml) No. of Trial Very Active
24.40

Table 6 shows the negative control is inactive and the positive control and the

flavon0id extract concentrations of 250g/ml, 500mg/ml, and 100mg/ml had a very

active result that means that it had a potential antibacterial activity.

 Table 7

Results of Zone of Inhibition by the Treatment of Semi-Purified Flavonoid Extract

on Pseudomonas aeruginosa

Result Zone of Interpretation

(mm) Inhibition in
(mm)
Negative Control Mean 0 Inactive
No. of Trial 0 0
0
Positive Control Mean 22.70
No. of Trial 24.40 23.07 Very Active
22.10
Flavonoid extract Mean 15.15
(250mg/ml) No. of Trial 15.10 15.22 Active
15.40
Flavonoid extract Mean 16.80
(500mg/ml) No. of Trial 16.40 16.52 Active
16.35
Flavonoid extract Mean 17.40
(1000mg/ml) No. of Trial 17.30 17.12 Active
16.65

Table 7 shows that the negative control is inactive and the positive control is

very active. The flavonoid concentrations of 250mg/ml, 500mg/ml, and 1000mg/ml

exhibited active result.

 Table 8

Results of Zone of Inhibition by the Treatment of Semi-Purified Flavonoid Extract

on Bacillus subtilis

Result Zone of Interpretation

Inhibition in
(mm)
(mm)

Negative Control Mean 0

No. of Trial 0 0 Inactive
0

Positive Control Mean 22.50

No. of Trial 22.60 22.43 Very Active
22.20

Flavonoid extract Mean 23.50

(250mg/ml) No. of Trial 21.40 22.1 Very Active
21.40

Flavonoid extract Mean 23.10

(500mg/ml) No. of Trial 21.40 22.73 Very Active
23.70

Flavonoid extract Mean 23.70

(1000mg/ml) No. of Trial 22.40 22.7 Very Active
22.30

Table 8 shows that the negative control is inactive and the positive control and the

concentration of the flavonoid ranging from the concentrations 250mg/ml, 500mg/ml,

and 1000mg/ml show a very active result having a potential of antibacterial activity.
4. Microbiological Test of the Flavonoid Extract in Liquid Handsoap

Table 9

Results of Zone of Inhibition by the Treatment of the Flavonoid Extract in Liquid

Handsoap on Escherichia coli

Result Zone of
Inhibition in Interpretation
(mm) (mm)

Negative Mean

Control No. of Trial 0 0 Inactive

Positive Mean

Control No. of Trial 25.35 25.35 Very Active

Mean
Flavonoid 9.60
extract in No. of Trial
10.25
liquid Hand 9.75 Inactive
soap 9.50

Table 9 shows that the flavonoid extract in antibacterial handsoap treated to

Escherichia coli is inactive.

 Table 10

Results of Zone of Inhibition by the Treatment of the Flavonoid Extract in Liquid

Handsoap on Staphylococcus aureus

Result Zone of
Inhibition in Interpretation
(mm) (mm)

Negative Mean

Control No. of Trial 0 0 Inactive

Positive Mean

Control No. of Trial 19.75 19.75 Very Active

Mean
Flavonoid 23.50
extract in No. of Trial
19.30
liquid Hand 20.28 Very Active
soap 18.05

Table 10 shows that the flavonoid extract in antibacterial handsoap treated to

Staphylococcus aureus is very active.

 Table 11

Results of Zone of Inhibition by the Treatment of the Flavonoid Extract in Liquid

Handsoap on Pseudomonas aeruginosa

Result Zone of
Inhibition in Interpretation
(mm) (mm)

Negative Mean

Control No. of Trial 0 0 Inactive

Positive Mean

Control No. of Trial 19.05 19.05 Very Active

Flavonoid Mean 28.60

extract in
No. of Trial 30.15 Very Active
liquid Hand 28.08
soap 25.50

Table 11 shows that the flavonoid extract in antibacterial handsoap treated to

Pseudomonas aeruginosa is very active.

 Table 12

Results of Zone of Inhibition by the Treatment of the Flavonoid Extract in Liquid

Handsoap on Bacillus subtilis

Result Zone of
Inhibition in Interpretation
(mm) (mm)

Negative Mean

Control No. of Trial 0 0 Inactive

Positive Mean

Control No. of Trial 24.15 24.15 Very Active

Mean
Flavonoid 27.90
extract in No. of Trial
16.05 23.02 Very Active
liquid Hand
soap 25.10

Table 12 shows that the flavonoid extract in antibacterial handsoap treated to

Bacillus subtilis is very active.

 STATISTICAL TREATMENT
Extract

93.37 ±16.22 100.36 ±4.25

70.58 ±3.81

42.36 ±15.76

There is a statistically significant difference between groups as determined by

One-way ANOVA, F (3,32)= 45.29, p= 0.000. A Tukey Post-hoc test revealed

that the relative inhibition zone is statistically highest on B. subtilis (100.36

±4.25, p= 0.000) and S. aureus (93.37 ±16.23, p= 0.000) compared to E. coli

(42.36 ±15.76, p= 0.000) and P. aeruginosa (70.58 ±3.81). Homogenous subsets

of the relative inhibition on different bacteria indicate which bacteria are

significantly different from each other. Three different subsets were observed,

which implies that the relative inhibitions on S. aureus and B. subtilis are

statistically different from both E. coli and P. aeruginosa.

Soap

147.42 ±12.43

95.31 ±25.65
102.70 ±14.46

38.59 ±1.60

There is a statistically significant difference between groups as determined by One-way

ANOVA, F (3,8)= 23.38, p= 0.000. A Tukey Post-hoc test revealed that the relative

inhibition zone is statistically highest on P. aeruginosa (147.42 ±12.43, p= 0.017) and

S. aureus (102.70 ±14.46, p= 0.037) compared to E. coli (38.59 ±1.60, p= 0.011) and B.

subtilis (70.58 ±3.81). Homogenous subsets of the relative inhibition on different

bacteria indicate which bacteria are significantly different from each other. Three

different subsets were observed, which implies that the relative inhibitions on S. aureus

and B. subtilis are statistically different from both E. coli and P. aeruginosa.