A.

DETERMINATION OF PH
INTRODUCTION
Fish is consumed as food all over the world. It is generally recognized as
polyunsaturated fatty acids. The composition might vary among species of fish. To
determine the pH of fish, it can be measure using an electrode and pH meter. The
fish mixtures were directly measured for pH. Different fish has different pH and it is
around natural pH. The pH of the fish also is affected by many factors such as type
of fish, flesh, habitat and etc.
OBJECTIVE
To determine the pH of the fish
PROCEDURE
1.
2.
3.
4.

The pH meter is calibrated with pH buffer 4 and 7

5 g of fish is grinded with addition of 5 mL of distilled water
The fish mixture is put into a beaker
The pH meter probe is inserted into the beaker and the pH is recorded

DISCUSSION
From the experiment, we need to determine the pH of the fish which is sardine and
kembung. The pH of fish is determined by using pH meter with pH buffer 4 and 7.
The fish is grinded with 5 ml of distilled water before the pH is determined. Besides,
pH is a measure of the acidic or alkaline nature of a solution. The concentration of
the hydrogen ion activity in a fish will determine the pH. Different fish also will result
different pH. Mostly, the pH of fish is around natural pH which is in range 6 9.
Through the result, the pH of kembung and sardine fish is determined which are
6.19 and 5.60 respectively. The moisture content of different type of fish also is
determined by using moisture analyzer which result 76.76 for kembung and 77.83
for sardine fish. The pH measurement and the moisture content determination of
fish also can give a good indication of the keeping qualities and freshness of the fish
product. Besides, the pH of fish also can be affected by rigor mortis process that
occurs after the death of fish where the glycogen in fish muscle is converted to
lactic acid. This process may lower the pH of the fish in order to prevent microbial
spoilage.
During the experiment, we also did some error that may affect the accuracy of the
result such as the fish is exposed to the air for longer period of time as it may
oxidize before the pH is determined. Moreover, the pH meter also may not calibrate
properly as we not use distilled water to rinse the probe meter that may interrupt
the result. Hence, to minimize these errors various precautions should be taken. For

example, three consecutive of checking pH and weighing the samples were carried
out to eliminate the errors of the handlers.
RESULT
FISH
KEMBUNG
SARDINES

PH
6.19
5.60

MOISTURE
76.76
77.83

CONCLUSION
As a conclusion, the pH of the fish is varies according to their species and also
habitat. The pH of fish is also around natural pH which is 6 9. Hence , the pH of
kembung and sardines fish is determined which are 6.19 and 5.60 respectively. The
hypothesis is accepted.

B . ASSESSMENT OF LIPID OXIDATION BY THIOBARBITURIC ACID METHOD ( TBA )

INTRODUCTION
The thiobarbituric acid (TBA) test is a method to detect oxidative deterioration of
fat-containing foods. During lipid oxidation, malonaldehyde (MA) is formed as a
result of the degradation of polyunsaturated fatty acids. It is usually used as an
indicator of the lipid oxidation process. In this experiment, the MA is reacted with
thiobarbituric acid (TBA) to form a pink MA-TBA complex that is measured
spectrophotometrically at its absorption maximum at 530535 nm. On the other
hand, lipid peroxidation is referring to the oxidative degradation of lipid. It is the
process in which free radical steal electrons from the lipid in cell membranes. This
process proceeds by a free radical chain reaction mechanism. Lipid oxidation is one
of the major causes of quality deterioration in fish. Thus, an increases in TBA
number also indicates an increases of measure in lipid peroxidation.

OBJECTIVES
To determine the TBA number in fish
PROCEDURE
1. 10 g of fish is macerate with 50 mL distilled water for 2 minutes and washed
into a distillation flask with 47.5 mL distilled water
2. 2.5 mL of 4 M HCL is added to bring the pH to 1.5

3. The distillation flask is heated by means of distillation apparatus so that 50

mL distillate is collected from the time boiling commences
4. 5 mL distillate is pipetted into a glass stopper tube and 5 mL of TBA reagent
is added. Then it is shaked and heated in boiling water for 35 minutes.
5. The tubes is cooled in water for 10 minutes
6. The blank similarly using 5 mL distilled water with 5 mL TBA reagent is
prepared
7. The absorbance is measured against the blank at 538 nm using 1 cm cells

DICUSSION
From the experiment, we need to determine the TBA number in different fish which
are Indian mackerel (kembung) and sardines. TBA is a method to detect oxidative
deterioration of fat-containing foods. Besides, it is performed to measure the
amount of MDA present in the fish. During lipid oxidation, malonaldehyde (MA) is
formed as a result of the degradation of polyunsaturated fatty acids. It is usually
used as an indicator of the lipid oxidation process. In this experiment, the MA is
reacted with TBA with absorption of 538 nm. The method involved heating of fish
with TBA reagent for approximately 35 minutes in boiling water bath. After cooling,
the solution is centrifuged for 20 min. The absorbance was then determined at 538
nm against a blank and the TBA number is calculated.
Through the experiment, the absorbance and TBA number of both kembung and
sardines is determined. The absorbance for kembung fish is 0.146 and sardines
0.174. While, the TBA number for both kembung and sardines is calculated which
result 1.1388 mg/kg and 1.3572 mg/kg respectively. The TBA number for sardines is
slightly higher than kembung because sardines contain higher amount of lipids and
myoglobin in the dark muscle cell. Fish that has dark flesh may oxidize rapidly than
fish that has white flesh because of the higher amount of Fe in myoglobin which can
accelerates the oxidative process. An increased in TBA value is also can be
measured as the extent of oxidative deterioration in fish.
In addition, the commercial value of both fish also can be determined by using TBA
method which sardine fish has lower commercial value compared to kembung. It
was proved in food industry which fish which have white flesh has higher rate in
industry than dark flesh fish. However, during the experiment, there are some errors
that we done that may affect the accuracy of the result such as the sample has
been exposed to the air for longer period of time. It may cause the fish to be
oxidized rapidly. On the other hand, the apparatus provided is not enough as we
change the distillation method to boiling method in order to conduct this

experiment. Besides, the blank also not prepared properly and some of foreign
particles in the sample tube are detected as it may interrupt the reading of the
absorbance of both fish.

RESULT

FISH
SARDINES
KEMBUNG

ABSORBANCE ( D )
0.174
0.146

TBA NUMBER
0.174 X 7.8 = 1.3572 mg/kg
0.146 X 7.8 = 1.1388 mg/kg

CONCLUSION
In this experiment, the TBA number of both kembung and sardine fish can be
determined which are 1.1388 and 1.3572 respectively. It is proved that TBA method
can be used to determine the oxidative rancidity of certain food especially fish.
Hence, the hypothesis is accepted.

REFERENCES
Fereidoon Shahidi and Ying Zhong , ( 2000 ) , Lipid Oxidation: Measurement
Method
,
Retrieved
from
http://uqu.edu.sa/files2/tiny_mce/plugins/filemanager/files/4281709/84607_08.pdf
Measurement
of
Lipid
Oxidation
,
(
n.d
)
,
Retrieved
from
http://www.public.iastate.edu/~duahn/teaching/Lipid%20oxidation/Measurement
%20of%20Lipid%20Oxidation.pdf
MARK P. RICHARDS , ( 2004 ) , Contributions of Blood and Blood Components to
Lipid
Oxidation
in
Fish
Muscle,
Retrieved
from
http://www.aseanfood.info/Articles/11019516.pdf
Hans H. Huss , ( 1988 ) , Fresh Fish - Quality and Quality Changes , FAO/DANIDA
agencies

1)

What are the methods that can be used as a spoilage indicator in fish?

Methods by detection of CO2 using an infrared CO2 analyzer or volatile amines like
Trimethylamine involves reduction of TMAO to TMA. *ak xpuas hati sgt*
2)

List the types of spoilage in fish.

Enzymatic autolysis, oxidation and microbial growth.#

3)

How is TBA associated with the quality of fish?

Using TBA for determination of the total lipid content in fish meat could associated
with the quality of fish by omitting the addition of antioxidant to the reaction
mixture during TBA reaction. It can be a good indicator of rancidity, particularly for
fish products.