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1. KENDRIYA VIDYALAYA SANGATHAN
SESSION 2022-2023
CLASS - XII SCIENCE
CHEMISTRY INVESTGATORY PROJECT
IRON CONTENT IN APPLE JUICE
SUBMITTED BY:
Name: VRIDDHI KESHODHAN
School: KENDRIYA VIDYALAYA SANGATHAN
I SHIFT

2. CERTIFICATE
This is to certify that Vriddhi Keshodhan of class
XII of KENDRIYA VIDYALAYA SANGATHAN has
successfully completed her project report in
chemistry on the topic
“Iron Content in Apple Juice”
for the partial fulfillment of AISSCE as prescribed
by the CBSE in the year 2022-2023.
Signature of the Signature of the
Subject Teacher Examiner

3. ACKNOWLEDGEMENT
At the outset, I express our gratitude to the
Almighty Lord for the divine guidance and wisdom
showered on me to undertake this project.
I am immensely grateful to my Principal for his
involvement in this project by providing useful
inputs and timely suggestions.
I am also thankful to my Chemistry teacher Mrs.
Jyotsna Dwiwedi for her guidance and help to make
this project a success.
My parents also played a key role in shaping up this
project nicely and I convey my special thanks to
them as well.

4. CONTENTS
➢ Objective
➢ Introduction
➢ Iron
➢ Sources of Iron
➢ Amount of Iron Intake
➢ Apple Juice
➢ Health Benefits of Apple Juice
➢ Test for Iron
➢ Estimation of Iron by:
- Spectrophotometric method
- Reagent used
- Sample preparation
- Standard Preparation
➢ Result and Discussion
➢ Conclusion

5. ANALYSIS OF
IRON CONTENT
IN APPLE JUICE

6. OBJECTIVE
To analyse apple juice for the presence of iron content
in it
INTRODUCTION
Iron is an essential element for most life on Earth,
including human beings. About 70 percent of
your body’s iron is found in the red blood cells of your
blood called hemoglobin and in muscle cells called
myoglobin.
Iron is needed for a number of highly complex
processes that continuously take place on a molecular
level and that are indispensable to human life, e.g. the
transportation of oxygen around your body.
Iron is required for the production of red blood cells (a
process known as haematopoiesis), but it is also part
of haemoglobin (that is the pigment of the red blood
cells) binding to the oxygen and thus facilitating its
transport from the lungs via the arteries to all cells
throughout the body.
The production of enzymes (which play a vital role in
the production of new cells, amino acids, hormones
and neurotransmitters) also depends on iron.
The immune system is dependent on iron for its
normal functioning. Iron also contributes to normal
cognitive function in children.

7. IRON [Fe]
Iron is a chemical
element with symbol Fe (from Latin: ferrum)
and atomic number 26. It is a metal that belongs to
the first transition series and group 8 of
the periodic table. It is, by mass, the most common
element on Earth, right in front of oxygen (32.1%
and 30.1%, respectively), forming much of
Earth's outer and inner core. It is the fourth most
common element in the Earth's crust.
Chemically, the most common oxidation states of
iron are iron(II) and iron(III). Iron shares many
properties of other transition metals, including the
other group 8 elements, ruthenium and osmium.
Iron forms compounds in a wide range of oxidation
states, −2 to +7. applications.
Examples of iron-containing proteins in higher
organisms include
hemoglobin, cytochrome (see high-valent iron),
and catalase. The average adult human contains
about 0.005% body weight of iron, or about four
grams, of which three quarters is in hemoglobin – a
level that remains constant despite only about one
milligram of iron being absorbed each day, because
the human body recycles its hemoglobin for the
iron content.

8. SOURCES OF IRON
Iron from food comes in two forms: heme and non-
heme. Heme is found only in animal flesh like meat,
poultry, and seafood. Non-heme iron is found in
plant foods like whole grains, nuts, seeds, legumes,
and leafy greens. Non-heme iron is also found in
animal flesh (as animals consume plant foods with
non-heme iron) and fortified foods.
Meats, poultry, and seafood are richest in heme
iron. Fortified grains, nuts, seeds, legumes, and
vegetables contain non-heme iron. In the U.S. many
breads, cereals, and infant formulas are fortified
with iron.
Bran fiber, large amounts of calcium particularly
from supplements, and plant substances
like phytates and tannins can inhibit the absorption
of non-heme iron.
Sources of non-heme iron:
• Beans
• Lentils
• Spinach
• Potato with skin
• Nuts, seeds

9. CAUSES AND CONSEQUENCES OF IRON
DEFICIENCY
Iron deficiency is defined as a condition in which
there are no mobilizable iron stores and in which
signs of a compromised supply of iron to tissues,
including the erythron, are noted. Iron deficiency
can exist with or without anemia. Some functional
changes may occur in the absence of anemia, but
the most functional deficits appear to occur with
the development of anemia. Even mild and
moderate forms of iron deficiency anemia can be
associated with functional impairments affecting
cognitive development, immunity mechanisms, and
work capacity. Iron deficiency during pregnancy is
associated with a variety of adverse outcomes for
both mother and infant, including increased risk of
sepsis, maternal mortality, perinatal mortality, and
low birth weight. Iron deficiency and anemia also
reduce learning ability and are associated
with increased rates of morbidity.[68]
Causes of iron deficiency
Iron deficiency results from depletion of iron stores
and occurs when iron absorption cannot keep pace
over an extended period with the metabolic
demands for iron to sustain growth and to replenish
iron loss, which is primarily related to blood loss.

10. The primary causes of iron deficiency include low
intake of bioavailable iron, increased iron
requirements as a result of rapid growth,
pregnancy, menstruation, and excess blood loss
caused by pathologic infections, such as hook worm
and whipworm causing gastrointestinal blood loss
and impaired absorption of iron. The frequency of
iron deficiency rises in female adolescents because
menstrual iron losses are superimposed with needs
for rapid growth. Other risk factors for iron
deficiency in young women are high parity, use of
an intrauterine device, and vegetarian diets.

11. AMOUNT OF IRON INTAKE
Infants and Children:
Younger than 6 months- 0.27/day
7 months to 1 year- 11mg/day
1 to 3 years old- 7mg/day
4 to 8 years- 10mg/day
Males:
9 to 13 years old- 8mg/day
14 to 18 years old- 11mg/day
19 and above- 8mg/day
Females:
9 to 13 years old- 8mg/day
14 to 18 years old- 15mg/day
18 to 51 years old- 15mg/day
51 and above- 8mg/day

12. HEALTH BENEFITS OF APPLE JUICE
Boosts Heart Health – Apples contain polyphenols
along with flavonoids. These polyphenols may
contribute towards combating the oxidation of bad
cholesterol or LDL and their subsequent build-up in
the arteries. Apple juice also contains potassium
which is good for the heart, reducing blood
pressure overall.
Relieves Asthma Symptoms – Apple juice helps
fight allergies and inflammation, relieving asthmatic
symptoms as a result. Its polyphenols contribute
towards enhancing the health of the lungs while
lowering pulmonary disease risks greatly.
Helps with Weight Loss – There are many
advantages of consuming apple juice for weight
loss. Apples come with carotenoids, polyphenols
and dietary fibre alike. Their juice may help in
lowering weight.
Reduces Cancer Risks – Phenolic acids and
flavonoids present in apple juice may effectively
work to combat cancerous cell development and
tumors as well. Apple juice may help in lowering
colon and lung cancer risks considerably.
Benefits the Eyes – Apple juice is rich in Vitamin A
that helps greatly in enhancing overall vision and
combating future disorders of the eyes as well.

13. EXPERIMENT
Apparatus: Test tube, burner, burner stand, test tube
holder, dropper
Materials Required: Apple juice, conc. nitric acid,
potassium sulphocyanide solution.
Procedure:
Step 1 - Take 2ml of apple juice in the test tube.
Step 2 – Add a drop of concentrated nitric acid (HNO3).
Step 3 - Boil the solution by mixing them well on the
burner.
Step 4 - Cool down the boiled solution and add 2-3
drops of potassium sulphocyanide solution.
Step 5 - (Observation) - Blood red colour is shown by
solution in the test tube. This blood red colour of the
solution indicates the presence of iron in the given
apple juice.
Result: Hence the given apple juice has iron content in
it.
Precautions:
• Mixture should be stirred continuously while
heating.
• Excess of reagents should not be used.

14. ESTIMATION OF IRON BY
SPECTROPHOTOMETRIC METHOD
CALIBRATION OF SPECTROPHOTOMETER:
Calibration of spectrophotometer was done by
standard method using potassium dichromate
solution.

15. REAGENTS USED
Aqua Regia:
HCl : HNO3
Hydrochloric acid : Nitric acid
3 : 1
0.1N KmnO4 : Dissolve accurately above 3.166 g of
KMNO4 powder in 100 ml distilled water. Warm if
necessary. Then cool to room temperature and
dilute to 1000 ml with distilled water.
4M nitric acid [HNO3] : 25% v/v nitric acid
Ammonium thiocyanate solution : A 10% w/v
solution of ammonium thiocyanate
Dilute sulphuric acid [H2SO4] : 5% v/v sulphuric
acid

16. SAMPLE PREPARATION
1 g of apple juice concentrate powder was
accurately weighed in silica crucible. The sample
was then ignited at 450 °C-550 °C for 10-12 hours
and cooled to room temperature. The residue was
then dissolved in 60-80 ml of aqua regia and then
boiled and concentrated to 10-15 ml on a hot plate.
The solution was cooled to room temperature and
diluted to 100 ml with distilled water.
After shaking the solution well, it was filtered
through Whatman filter paper no. 1. Now, 5 ml of
this filtrate was further diluted to 100 ml with
distilled water [Solution A] and 2-4 ml of solution
A was accurately pipetted into a 50 ml volumetric
flask for colour development.
Solution was prepared in a similar manner by
omitting the sample [Solution B].

17. STANDARD PREPARATION
About 703 mg of ferrous ammonium sulphate was
accurately weighed and dissolved in 100 ml distilled
water and then 5 ml dilute H2SO4 was added
dropwise until a pink colour persists. Then the
above volume was made up to 1000 ml with
distilled water. This solution C was well shaken and
left overnight. Next day, 1 ml of this solution was
pipetted out accurately for colour development.
Further dilutions were made as per the table.
Reagents Blank Standard Sample
Stock solution
[ml]
1 ml of solution
B
1 ml of solution
C
2 ml solution A
0.1 N KMnO4 Till persistent
pink colour
Till persistent
pink colour
Till persistent
pink colour
4M HNO3 [ml] 3 3 3
Distilled water 5 5 5
10% ammonium
thiocyanate
solution
10 10 10
Distilled water Dilute upto 50ml Dilute upto 50ml Dilute upto 50ml

18. The final dilutions were made up to 50 ml and
absorbance reading for sample and standard
solutions were recorded at a wavelength of 480 nm
after 5 mins on previously calibrated suitable
spectrophotometer against the blank solution
prepared as mentioned above. The concentration of
iron in such sample was calculated in percentage
w/w using the following formula:
Content of iron [%w/w] = [Sample absorbance/
standard absorbance] x [o.ooo1/60] x [100/weight
of sample in g] x dilution factor x 100

19. RESULT
The respective iron contains following by digesting
the corresponding ashes
Description Iron Content[%w/w]
Greyish brown coloured powder 15.97
Greyish brown coloured powder 13.63
Dark grey coloured powder 7.33
Dark brown coloured powder 6.90
Grey coloured powder 6.75
Brown coloured powder 3.72
Light brown coloured powder 1.22

20. CONCLUSION
Greyish brown coloured sample is found to be the
richest source of iron content of about 15.67%
whereas the lowest iron content of 0.74% since
Apple juice content concentrate is a natural source,
it also contains vitamin C. However, the content of
vitamin C varies, but due to the presence of
vitamin, the bioavailability and absorption of iron
increases. Thus, we get additional benefit of using
this natural source of iron as a supplement.
Hence, instead of using synthetically prepared
chemical forms of iron as supplements, it is
recommended to use apple juice concentrate, which
is a natural source of iron instead of supplements
and medicines.