FSG661/2016/Form A

PROGRESS REPORT FSG661

Week (circle) (4) (8) (12)
Name: Ahmad Faisal Bin Mohamad Roslan
Student No: 2014641722
Supervisor: Miss Lailatul Wahida Binti Musa
Title: Phytochemical Screening and Antimicrobial Activities of Methanolic and Hexanoic Extract
of Diospyros blancoi Leaf Towards Selected Gram Negative and Gram Positive Bacteria
Give a summary of what you did in the past four weeks. Provide data sample or results shown
in your FYP laboratory log book.
The project was continued with antimicrobial screening which is disks diffusion and MIC
determination. Disk diffusion was horrible at first when using hockey stick, using hockey stick
makes the streaking prone to contamination. The problem solved when using sterile cotton bud
for streaking. Using cotton bud is ideal because it is one time use and doesnt need to wait for
hockey stick cool down which is then can save a lot of time by doing so. To reduce the risk of
contamination, 5 stick of cotton bud was dressed with aluminium foil then autoclave it. The disk
diffusion testing was started with methanolic extract. It is working fine all of it forming zone of
inhibition except for P. aeruginosa. The disk diffusion method is then proceed with hexanoic
extract. None of the bacteria streaked form zone of inhibition. Each of the result was triplicated
and produced the same result.

Bacteria

Staphylococcus
aureus

Bacillus subtilis

Concentration
(mg/ml)

Sample Reading

Average
(Au)

1st

2nd

3rd

100
50
25
12.5

0.623
1.231
1.411
1.441

0.664
1.221
1.422
1.444

0.661
1.223
1.412
1.423

0.649
1.225
1.415
1.436

+ Control

0.322

0.331

0.22

0.291

- Control

1.422

1.432

1.444

1.432

100
50
25
12.5
+ Control

0.422
0.621
0.891
1.221
0.111

0.412
0.633
0.91
1.233
0.125

0.432
0.631
0.898
1.241
0.122

0.422
0.628
0.899
1.231
0.119

FSG661/2016/Form A

Pseudomonas
aeruginosa

Klebsiella
pneumoniae

- Control

1.321

1.332

1.328

1.327

100
50
25
12.5

2.223
2.211
2.23
2.117

2.111
2.243
2.244
2.104

2.117
2.233
2.112
2.121

2.15
2.229
2.195
2.114

+ Control

0.551

0.552

0.555

0.552

- Control

2.112

2.101

2.117

2.11

100
50
25
12.5

0.66
1.123
1.33
1.372

0.641
1.112
1.334
1.237

0.65
1.154
1.239
1.224

0.65
1.129
1.301
1.277

+ Control

0.332

0.331

0.302

0.321

- Control

1.345

1.354

1.332

1.343

FSG661/2016/Form A
The extract is then proceed with the re-screening process. The result is as below.

Phytochemical
Methanol

Hexane

Alkaloids

Flavonoids

Tannin

Terpenoids

Saponin

Constituents

The result obtained and shown as in the picture is differ from actual vision. Some of the
changes indicate the presence of each phytochemical cannot be seen in camera due to
lightning related exposure.
Minimal Inhibitory Concentration (MIC) was determined by using manual method, by using UVVis spectroscopy. The result maybe not reliable as expected due to lacking apparatus.

FSG661/2016/Form A

Bacteria

Staphylococcus
aureus

Bacillus subtilis

Pseudomonas
aeruginosa

Klebsiella
pneumoniae

Concentration
(mg/ml)

Sample Reading

Average
(Au)

1st

2nd

3rd

100
50
25
12.5

0.623
1.231
1.411
1.441

0.664
1.221
1.422
1.444

0.661
1.223
1.412
1.423

0.649
1.225
1.415
1.436

+ Control

0.322

0.331

0.22

0.291

- Control

1.422

1.432

1.444

1.432

100
50
25
12.5

0.422
0.621
0.891
1.221

0.412
0.633
0.91
1.233

0.432
0.631
0.898
1.241

0.422
0.628
0.899
1.231

+ Control

0.111

0.125

0.122

0.119

- Control

1.321

1.332

1.328

1.327

100
50
25
12.5

2.223
2.211
2.23
2.117

2.111
2.243
2.244
2.104

2.117
2.233
2.112
2.121

2.15
2.229
2.195
2.114

+ Control

0.551

0.552

0.555

0.552

- Control

2.112

2.101

2.117

2.11

100
50
25
12.5

0.66
1.123
1.33
1.372

0.641
1.112
1.334
1.237

0.65
1.154
1.239
1.224

0.65
1.129
1.301
1.277

+ Control

0.332

0.331

0.302

0.321

- Control

1.345

1.354

1.332

1.343

The MIC was determined when 50% of bacteria population is killed.

FSG661/2016/Form A
The statistical analysis was done by using two-way ANOVA by aid of IBM SPSS, EXCEL, and
Minitab. Three of them produced same result except that Minitab and Excel doesnt display
standard error for the analysis.

FSG661/2016/Form A
Signature of Students

Signature of Supervisor:

Name: Ahmad Faisal Bin Mohamad Roslan

Name: Miss Lailatul Wahida Binti Musa

Student No: 2014641722

Official Stamp: