ACKNOWLEDGEMENT

Firstly, I want to say thank you to En Zailuddin Ariffin as a lecturer of this course that

has spend lot of time to teach and observe my project experiment. For En Razali as Chemistry

Laboratory 1 Assistant that always spends a time to give me used a lab for done my project

experiment. Also to the Cik Naimah as Instrumental Laboratory Assistant that always teach me

to used a instrument and spend a lot of time to give me time for finished my project experiment.

Also I want to say thanks, to my family and to my entire friend for the morale support for me to

finish the project experiment in period of time.

ABSTRACT

This experiment was done to determine the present and amount of iron and zinc in fish

based on their habitat. Fish that has been choosing is catfish from pond, snake head fish from

river and mackerel from sea. This sample preparation was done by using dry ashing method. The

sample then has been analyzed by using Flame Atomic Absorption Spectroscopy (AAS). After

analyzed, the present of iron and zinc has been detected in an each sample. Based on the result,

mackerel has higher amount of iron and snake head fish has higher amount of zinc. Catfish has

lowest amount of both iron and zinc. Based on this result also, it can tell that the amount of iron

and zinc contain in this three sample is not exceeded the amount of toxicity that has been list by

Food and Drug Administration (FDA). So it’s considered safe for consumption.

1.0 INTRODUCTION
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 Iron, is one of the most valuable metal on earth, is essential to most life form and to

normal human physiology. Iron is an integral part of many protein and enzymes that maintain

good health. In human, iron is an essential component of protein involved in oxygen transport. It

is also essential for the regulation of cell growth and differentiation. A deficiency of iron limits

oxygen delivery to cell, resulting in fatigue, poor work performance and decreased immunity.

Large amounts of ingested iron can cause excessive levels of iron in the blood. High

blood levels of free ferrous iron react with peroxides to produce free radicals, which are highly

reactive and can damage DNA, proteins, lipids, and other cellular components. Thus, iron

toxicity occurs when there is free iron in the cell, which generally occurs when iron levels

exceed the capacity of transferring to bind the iron.

The mineral zinc is present in every part of the body and has a wide range of functions. It

helps with the healing of wounds and is a vital component of many enzyme reactions. Zinc is

vital for the healthy working of many of the body's systems. It is particularly important for

healthy skin and is essential for a healthy immune system and resistance to infection. Zinc has a

range of functions. It plays a crucial role in growth and cell division where it is required for

protein and DNA synthesis, in insulin activity, in the metabolism of the ovaries and testes, and in

liver function. As a component of many enzymes, zinc is involved in the metabolism of proteins,

carbohydrates, lipids and energy.

Fish is an example of food source that provides iron and zinc. In this experiment, the

amount of iron and zinc in fish will be determined. With this experiment, it can tell whether fish

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now day is safe to eat or not. This experiment also, has used three kinds of fish that will be

analyzed as sample based from their habitat it is pond, sea and river. Why it based on fish

habitat? It is because, different habitat they consume different way to live. With this different

way, the amount of iron and zinc in fish will be different.

Fish that has been breed in pond has a potential to provide excess iron and zinc. It is

because, in their fish food contain some trace element like iron and zinc. Fish from river also has

potential in this case. Now days, the river has been polluted. Heavy metal and trace element exist

in river. So it will be affected the fish with increasing amount of iron and zinc in fish immune

system. Fish came from sea also can be said not very safe to eat. A sea also faces a crisis of

pollution. This has been affected the aquatic life with toxicity of trace element and heavy metal.

In this experiment, Flame Atomic Absorption Spectroscopy is been used to detect the

presence of iron and zinc. Flame atomic absorption is a very common technique for detecting

metals and metalloids in environmental samples. It is very reliable and simple to use. The

technique is based on the fact that ground state metals absorb light at specific wavelengths. Metal

ions in a solution are converted to atomic state by means of a flame. Light of the appropriate

wavelength is supplied and the amount of light absorbed can be measured against a standard

curve.

2.0 OBJECTIVE

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To determine the present and amount of Iron and Zinc in fish based on their habitat (Sea, pond,

river) by using Flame Atomic Absorption Spectroscopy (AAS).

3.0 APPARATUS

• 50ml volumetric flask

• 100ml volumetric flask

• Burette

• 150ml beaker

• 10ml measuring cylinder

• Filter funnel

4.0 CHEMICAL

• 1000ppm zinc stock solution

• 1000ppm iron stock solution

• 65% concentrated Nitric Acid (HNO3)

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5.0 PROCEDURE

5.1Preparation of standard for Zinc and Iron

10ml of 1000ppm stock solution was measured. Then, it was transferred into a

100ml volumetric flask and was diluted using deionised water to make it be 100ppm

stock solution. Burette was used to transfer this 100ppm stock solution to prepare a

standard of 0.5ppm, 1.0ppm, 1.5ppm, 2.0ppm, 2.5ppm. Ratio of standard is 1ml=1ppm,

so for 0.5ppm transferred 0.5 ml of stock solution into a volumetric flask, for

1.0ppm(1ml), 1.5ppm(1.5ml), 2.0ppm(2.0ml), 2.5ppm(2.5ml). Deionised water was used

to dilute all standard. All standard then was transferred into a sample bottle to analyses

5.2Preparation of sample

Fish was washed cleanly by used tap water. Fish head has been cut and fish meat

was separated from bone. Then, fish was blend by using electric blender. (small amount

of water was poured) Then, sample was weighed about 20 g in a crucible. Then, sample

was burned in a furnace at 550oC for 90 minutes. After that, 5ml of 65% concentrated

Nitric Acid (HNO3) was measured and was transferred into a crucible containing ash

sample to dissolved ash sample. Then, sample was transfer into a beaker containing small

amount of deionised water. Then, sample was filtered by using filter paper and filter

funnel into a 50ml volumetric flask. Then, sample was diluted until the mark by using

deionised water. Then, sample was transferred into a sample bottle for analyses.

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 5.3Flame AAS manual

i. Compressor air, stopcock air, acetylene gas, isolethene switch, exzos fan, instrument

and PC have been switch on and open.

ii. For analyses, Winlab 32(AA) software was used.

iii. Then, click file, new method, choose element, method description (title), then click

calibration, choose linear through zero, unit ppm (w/v), sample ppm (w/v)

iv. Then click standard concentration, calibration blank, fill up the space, then click save

as, method, ok.

v. Click file, new sample info design file, default, ok, batch ID (date), sample ID, file,

save as sample info file.

vi. Click tools, flame control, manual analysis, and calibration display, edit calibration.

vii. Result data, set name.

viii. Open flame, then click analyze blank.

ix. Then, put standard, and click analyze standard.

x. After, get the best line that correlation factor is 0.999 or 0.9999, run the sample by

click analyze sample.

xi. Print all result obtained.

xii. Then switch off all steps in i.

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6.0 OBSERVATION

i. After burned in furnace, the sample ash looks grey-white in color.

ii. Then after dissolved with concentrated Nitric acid (HNO3), it become black solution.

iii. Then after filter and dilute in 50ml volumetric flask, solution become clear less.

iv. When sample run with flame AAS, the flame color from blue-orange color turn to

bright orange color means that the present of metal in sample.

Sample Weight of trial i Weight of trial ii Weight of trial iii

Catfish 20.0018 20.0007 20.0009

Snake head fish 20.0019 20.0017 20.0021

Mackerel 20.0012 20.0022 20.0015

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7.0 RESULT AND DATA

7.1 Table of data (Iron) from result

Sample Trial 1 Trial 2 Trial 3

Catfish 0.987ppm 0.982ppm 1.079ppm

Mackerel 2.166ppm 2.124ppm 2.051ppm

Snake head fish 1.308ppm 1.262ppm 1.178ppm

Ppm

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2.5

1.5
Trial 1
Trial 2
1
Trial 3
0.5

0
Catfish Mackerel Snake head

7.2 Table of data (Zinc) from result

Sample Trial 1 Trial 2 Trial 3

Catfish 1.719ppm 1.671ppm 1.692ppm

Mackerel 2.215ppm 2.364ppm 2.211ppm

Snake head fish 2.278ppm 2.251ppm 2.327ppm

ppm

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 2.5

1.5 Trial 1
Trial 2
1
Trial 3
0.5

0
Catfish Mackerel Snake Head

8.0 CALCULATION

8.1For standard range calculation

10ml x 1000ppm = 100ppm

100ml

So the ratio for standard dilution is 1ppm = 1ml

8.2Calculation for change ppm (w/v) to ppm (w/w)

Formula:
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 Concentration from result x volume dilute in L

Mass of sample weigh in kg

For Catfish Trial I (Iron) :-

0.987 mg / L x 0.05 L = 2.468 mg/kg

0.02 kg

8.3Table of data (Iron) in mg/kg

Sample Trial 1 Trial 2 Trial 3

Catfish 2.468 2.455 2.698

Mackerel 5.415 5.311 5.127

Snake head fish 3.272 3.155 2.945

8.4Table of data (Zinc) in mg/kg

Sample Trial 1 Trial 2 Trial 3

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 Catfish 4.292 4.185 4.231

Mackerel 5.546 5.911 5.533

Snake head fish 5.691 5.632 5.812

9.0 DISCUSSION

Based on the data obtained, mackerel shown the highest result in iron and zinc. Catfish

shown lowest result for the both metal and snake head fish shown highest result in zinc and

moderate result in iron. Mackerel is a fish from the sea. From information obtained, at 3.5% of

salinity, the concentration of Iron and Zinc in sea water is 0.0034ppm and 0.0050ppm. So, the

probability of mackerel can consume higher amount of Iron and Zinc based on their habitat is

high. It can came from their food sources and by their respiration process that made mackerel

can ingested the sea water that through their mouth and gills. Catfish is a fish that has been breed

from pond. The lowest concentration of Iron and Zinc in this fish maybe because of the

composition of fresh water that has small concentration of this two metal and their food source

do not contain or contain small amount of this two metal. From another theory also, I assume

that maybe the catfish has naturally contain lowest concentration of this two metal. Snake head

fish is a fish from river. The highest concentration in Zinc maybe because from the river water

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that has been used for irrigation for planting and orchard has been expose for this metal that

actually has been used as micronutrient for plant.

Based on the Food and Drug Administration (FDA) the amount of intake for Zinc and

Iron in a day is 14mg for adult and 9mg for children for Zinc and 18mg for adult and 10mg for

children for Iron. So from the analysis result, the amount of Iron and Zinc in each 0.02kg of

sample is less than (FDA) intake suggestion. So this amount is considered as safe amount contain

in fish because do not exceed the limit of toxicity.

10.0 CONCLUSION

As conclusion, the present of Iron and Zinc in all three samples have been detected. The

concentration obtained from the analysis of these three samples shown it not exceeded the

toxicity limit. Also, from the analysis, these three samples have shown different concentration

for each sample. This makes sense, that habitat of fish also can affect the amount of Iron and

Zinc. So, the amount of Iron and Zinc in Catfish, Mackerel, and Snake Head fish is considered

safe for consumption.

11.0 REFERENCE

i. Determination of heavy metals in fish tissue, water and sediment from Epe and

Badagry lagoons, Lagos, Nigeria. Retrieve: 27th September 2010,7(1),215-212

http://www.e-journals.net

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 ii. Dr J Floor Anthony, The chemical composition of seawater. Retrieve: 12th October

2010, http://www.seafriends.org.nz./oceano/seawater.htm

iii. Iron, Retrieve: 19th October 2010, http://www.wikipedia.org

iv. Zinc, Retrieve: 19th October 2010, http://www.wikipedia.org

v. Plant nutrient, Retrieve: 19th October 2010, http://www.wikipedia.org

12.0 APPENDIX

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