Effect of different Acidity and

pickling time on microbiological,

physicochemical, and sensory
acceptance of Melon manis
terenggganu pickles

LEE CHYE YIEN

S59196
 CHAPTER 3
MATERIALS AND METHOD
3.1.2 METHOD OF PICKLING MELON MANIS TERENGGANU (MMT)
1. Fresh MMT are cut into pieces and washed with chlorinated water.
2. 245 g of fresh MMT are weighed and steeped in brine solution for 3 days in the
glass jars.
3. After 3 days, the brine solution are drained and pickling solution are prepared for pickling MMT.
4. Vinegar, sugar, water, and calcium chloride are added.
5. MMT pickles is pasteurised until the middle of the jar reaches 74°C.
6. The jars are sealed and placed in boiling water for 15 minutes, followed by 30 seconds of
tempering with lukewarm water.
7. Cooled the pickles under running tap water.
8. MMT are pickled for 5, 7, and 10 days.
Pickling solution with 2% Pickling solution with 3%
acidity acidity

70 g Vinegar 105 g Vinegar

105 g Water 70 g Water

80 g Sugar 80 g Sugar

1/16 tsp Calcium Chloride Pickling solution with 2.5% 1/16 tsp Calcium Chloride
acidity (for control)

87.5 g Vinegar

87.5 g Water

80 g Sugar

1/16 tsp Calcium Chloride

Final Product of MMT pickles
 3.2
PHYSICHOCHEMICAL ANALYSIS
OF MELON MANIS TERENGGANU
(MMT) PICKLES
 3.2.1 PH ANALYSIS
1. 30 g of pickled MMT are blended into a paste consistency by using a
blender.
2. pH probe are submerged in the sample and pH of the paste are measured.
3. Readings of pH are recorded once the pH meter stabilises.
 3.2.2 TEXTURE ANALYSIS
1. Crunchiness and firmness of MMT pickles are measured by using TA.XTplus Texture
Analyzer with p/2 needle probe.
2. MMT pickles with 1.5 cm thick are placed centrally under the probe, then control the
system to lower the probe and commence the test.
3. Results are analyzed by software.
 3.2.3 COLOUR ANALYSIS
1. MMT pickles are placed in small zip lock plastic bag.
2. Colour of MMT pickles are measured by using chroma meter.
3. The results are recorded once the chroma meter stabilises.
3.2.4 ASCORBIC ACID ANALYSIS
1. Prepare the DCPIP solution.
2. Prepare the sample solution. 16 g of MMT pickles are weighted and grinded into paste form and
filtered it to get the sample solution.
3. 1.5 ml of DCPIP solution are pipetted into a test tube, sample solution are added drop by drop until
the colour changes to colourless. Colourless indicates the presence of ascorbic acid.

PROBLEMS:
1. However, the colour of DCPIP solution in the test tube
does not changes into colourless, it turns dark red
regardless of how much sample solution I apply.
2. By consulting with CoSV, the colour does not changes
to colourless may due to the strong acidic of the MMT
pickles. The strongly acidic sample could confuse the
determination of the end point since they does not
decolourise completely but remains pink.
 MODIFICATION
1. 20 g of MMT pickles are cut into small pieces and grinded into paste form by using mortar and
pestle.
2. The paste are filtered and obtained the liquid sample extract.
3. 10 ml of sample extract was mixed with 50 ml oxalic acid. The mixture was stirred and 10 ml of
the filtrate was taken to titrate with 2,6-dichlorophenolindophenol.
4. Titration was stopped when a distinctive rose-pink endpoint was appeared and last more than
5 seconds.
DCPIP solution
 3.2.5 PROTEIN ANALYSIS
1. 2 grams of MMT pickles sample are weighed approximately and put into a 100 ml Kjeltec sample tube. 2 tablets of Kjeltabs
Catalyst (Cu 3.5) are added.
2. 12 ml of H2SO4 are added carefully and slightly shake the digestion tube. The tube are connected with Exhaust system
and the digestion process started. The digestion process is completed when the solution in the digestion tube change
color into light blue.
3. The tube are cooled down (20 minutes) and 75 ml of distilled water are added. To prepare acceptor solution, 25 ml of 4%
Boric acid are added with 10 drops of Green Bromocresol indicator in conical flask.
4. 50 ml of 40% sodium hydroxide are added into the tube. Distillation process started, the acceptor solution are added
inside distillation process. Distillation process continue for 4 minutes until the acceptor indicator changed color to light
green.
5. Titrated the acceptor indicator with 0.1 N HCI until color changed to grey.
6. Titration volume are recorded, calculated the crude protein content (%) using formula.
 3.3
MICROBIOLOGICAL ANALYSIS OF
MELON MANIS TERENGGANU
(MMT) PICKLES
3.3.1 PREPARATION OF AGAR MEDIUM

PCA
1. 500 ml of PCA agar are prepared by dissolving 8.75 g of PCA agar powder
with 500 ml of distilled water in a 1000 ml botol.
2. The agar solution are then autoclaved, cooled, and poured into the agar
plate.

PDA
1. 500 ml of PDA agar are prepared by dissolving 19.5 g of PDA agar powder
with 500 ml of distilled water in a 1000 ml botol.
2. The agar solution are then autoclaved, cooled, and poured into the agar
plate.
 BPA
1. 500 ml of BPA agar are prepared by dissolving 31.5 g of BPA agar powder
with 475 ml of distilled water in a 1000 ml botol.
2. The agar solution are then autoclaved and cooled down. 25 ml of Egg Yolk
Tellurite Emulsion are added.
3. The solution are poured into the agar plate.

VRBD
1. 500 ml of VRBD agar are prepared by dissolving 19.75 g of VRBD agar
powder with 500 ml of distilled water in a 1000 ml botol.
2. The solution are heated on heater until boiling.
3. The agar solution are then cooled and poured into the agar plate.
3.3.1.1 PREPARATION OF SALINE WATER & BPW

SALINE WATER
1. 700 ml of saline water are prepared by dissolving 5.95 g of sodium chloride with
700 ml of distilled water.
2. 9 ml of the saline solution are pipetted into a universal botol. The process are
repeated until all the saline water are finished pipette into the universal botol.
3. Then, autoclave it and stored for later used.

BUFFERED PEPTONE WATER (BPW)

1. 500 ml of BPW are prepared by dissolving 0.5 g of BPW powder with 500 ml of
distilled water.
2. 90 ml of the saline solution are measured by using measuring cylinder and poured
into a botol.
3. Then, autoclave it and stored for later used.
3.3.2 Spread plate method

PCA, PDA, VRBD, BPA

1. 10 grams of MMT pickles sample are cut and weighed approximately and put into a stomacher bag. 90 ml of BPW
are added into it and homogenized it.
2. PCA, PDA, VRBD and BPA agar plate are labelled. L-shaped hockey stick, alcohol, micropipette, micropipette tips
and bunsen burner are prepared.
3. For 10-1 dilution, 1000 μl of the solution from stomacher bag are pipetted and mixed it with 9 ml of saline water.
For 10-2, 1000 μl of the solution from 10-1 are pipetted and mixed with 9 ml of saline water. The process are
repeated for 10-3, 10-4, and 10-5 dilution.
4. 100 μl of the sample solution are pipetted and transferred to the agar plate. L-shaped hockey stick are dipped
with alcohol and heated to sterilize it. The liquid sample are spreaded evenly over the whole surface of the
medium.
5. The process are repeated for all PCA, PDA, VRBD, and BPA agar plate. The plates are inverted in the incubator at
30 - 37℃.
6. Observed the results of PCA agar plate after 24 hours, BPA and VRBD agar plate after 48 hours, whereas PDA agar
plate after 5 days.
 Labelled the agar plate. Prepared micropipette,
Cooled down and poured micropipette tips, saline water, L-shaped hockey
Prepared agar the solution into agar plate stick, bunsen burner, and alcohol
solution

Incubated the plates The sample are spreaded Vortexed the dilution
evenly on the medium
 3.4
SENSORY TEST OF MELON MANIS
TERENGGANU (MMT) PICKLES
 SENSORY ACCEPTANCE TEST FOR MMT PICKLES
1. All of the samples will be coded with 3 digits number.
2. Tray, sticker label, master sheet, qr code (for google form), tissue and rinsing cup are
prepared. Mutation was doned by followeing the master sheet.
3. 30 untrained panelists from University Malaysia Terengganu are randomly chosen for this
sensory evaluation test.
4. Free gifts are provided to the panelists.
 CHAPTER 4
RESULTS AND DISCUSSION
4.1 Results of Physicochemical Analysis
4.1.1 pH analysis of MMT pickles

Results:

From the results, it can be seen that MMT pickles with 2% acidity and 5 days of pickling
time has the highest pH level (less acidity). On the other hand, MMT pickles with 3%
acidity and 10 days of pickling time has the lowest pH level (stronger acidity).
 4.1.2 Texture analysis of MMT pickles

For both replication 1 and 2, it can be seen that MMT pickles with
pickling time of 7 days (2.5% acidity) has the highest firmness.
However, in term of crunchiness, Replication 1 MMT pickles with
pickling time of 5 days (2% acidity) has the highest crunchiness (0.37),
while in Replication 2 MMT pickles with pickling time of 7 days (2.5%
acidity) has the highest crunchiness (0.34).
 4.1.3 Colour analysis of MMT pickles
Results:

By comparison, the data

shows that the raw MMT is
brighter than pickled MMT in
terms of colour (L*).
 4.1.4 Ascorbic acid analysis of MMT
pickles
Results:

The amount of DCPIP required to change the color of oxalic acid mixture to rose pink for raw
MMT is the highest which is 0.4 ml - 0.45 ml. It means that raw MMT has more ascorbic acid
content than the pickled MMT.
The amount of DCPIP required to change the color of oxalic acid mixture to rose pink for
MMT pickles with 10 days pickling time (2% & 3% acidity) is the lowest which is 0.1 ml. It
means that MMT pickles with 10 days of pickling time has less ascorbic acid content.
 4.1.5 Protein analysis of MMT pickles
Amount of titration:
Nitrogen content and crude protein content
are calculated using formula:

Results:

Crude protein content of raw MMT

is the highest (0.744%- 0.832%), the
crude protein content decrease
after pickling.
 4.2 Results of Microbiological Analysis
4.2.1 PCA
REPLICATION 1:
For 5 days, 2% acetic acid: For 5 days, 3% acetic acid: For 7 days, 2.5% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 4.2.1 PCA
REPLICATION 2:
For 5 days, 3% acetic acid: For 7 days, 2.5% acetic acid:
For 5 days, 2% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 Colonies count of PCA
Replication 1:

Replication 2:
 4.2.2 BPA
REPLICATION 1:
For 5 days, 2% acetic acid: For 5 days, 3% acetic acid: For 7 days, 2.5% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 4.2.2 BPA
REPLICATION 2:
For 5 days, 2% acetic acid: For 5 days, 3% acetic acid: For 7 days, 2.5% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 Colonies count of BPA
Replication 1:

Replication 2:
 4.2.3 VRBD
REPLICATION 1:
For 5 days, 2% acetic acid: For 5 days, 3% acetic acid: For 7 days, 2.5% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 4.2.3 VRBD
REPLICATION 2: For 7 days, 2.5% acetic acid:
For 5 days, 3% acetic acid:
For 5 days, 2% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 Colonies count of VRBD
Replication 1:

Replication 2:
 4.2.4 PDA
REPLICATION 1:
For 5 days, 3% acetic acid: For 7 days, 2.5% acetic acid:
For 5 days, 2% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 4.2.4 PDA
REPLICATION 2:
For 5 days, 3% acetic acid: For 7 days, 2.5% acetic acid:
For 5 days, 2% acetic acid:

For 10 days, 2% acetic acid: For 10 days, 3% acetic acid:

 Colonies count of PDA
Replication 1:

Replication 2:
4.3 Results of sensory acceptance test