Aim of the Project

To study the presence of oxalate ion content in

guava fruit at different stages of ripening.

In this project, we will learn to

test for the presence of oxalate
ions in the guava fruit and how its
amount varies during different
stages of ripening.

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 Introduction
Guava is sweet, juicy and light or dark green coloured fruit. It is cultivated in all parts
of India. It is a common sweet fruit found in India and many other places around the
world. Guavas are plants in the Myrtle family (Myrtaceae), genus Psidium (meaning
“pomegranate” in Latin) which contains about 100 species of tropical shrubs. When ripe
guava acquires yellow colour and has penetrating strong scent. The fruit is rich in
vitamin C and minerals. It is also a rich source of oxalate and its content in the fruit
varies during different stages of ripening.
Guava fruit, usually 4 to 12 cm long, are round or oval depending on the species.
The outer skin may be rough, often with a bitter taste, or soft and sweet. Varying
between species, the skin can be of any thickness, is usually dark or light green before
maturity, but becomes yellow, maroon, or green when ripe. Guava fruit generally have
a pronounced and typical fragrance, similar to lemon rind but less sharp. The pulp of
the fruit may be sweet or sour, off-white cream in colour ("white" guavas) or deep pink
("red" guavas), with seeds of variable number and hardness embedded in the central
pulp, again depending on the species.

F
Fig. 1: White guavas Fig. 2: Red guavas

Besides its unique flavour and fragrance, guava has been hailed as one of the super
fruits due to the numerous health benefits it offers. It indeed is a powerhouse of
nutrients. This fruit is extraordinarily rich in vitamin C, lycopene and antioxidants that
are beneficial for skin. Guavas are also rich in manganese which helps the body to
absorb other key nutrients from the food that we eat. Guavas contain folate, a mineral
which helps promote fertility. The potassium in guavas helps normalize blood pressure
as well. Since it contains about 80% of water it helps keep our skin hydrated.

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Benefits of Guava
Guava has a high content of vitamin C – it has the highest content of vitamin C among
all the citrus fruits; one guava fruit contains four times more vitamin C than an average
size orange. It also contains ten times more vitamin A than a lemon, besides vitamin
B2, vitamin E, vitamin K, fibre, calcium, copper, folate, iron, manganese, phosphorous
and potassium. It also contains oxalates, amount of which varies depending on the
degree of ripening of the fruit. With all the nutrition that it provides, guava hardly
contains any fats. Moreover, it is one of the least chemically treated and sprayed fruits.
Some of the wider applications of guava are as follows.
1. Immunity booster: Vitamin C present in guava helps to improve immunity and
protects us against common infections and pathogens.
2. Lowers risk of cancer: Lycopene, quercetin, vitamin C and other polyphenols act
as potent antioxidants which neutralize free radicals generated in the body, thereby
preventing the growth of the cancer cells.
3. Diabetes-friendly: Due to rich fibre content and low glycaemic index, guavas
prevent the development of diabetes. While the low glycaemic index inhibits a
sudden spike in sugar levels, the fibre content ensures that the sugar levels are well
regulated.
4. Heart healthy: Guavas improve the sodium and potassium balance of the body,
thereby regulating blood pressure in patients with hypertension. Guavas also help
lower the levels of triglycerides and bad cholesterol (LDL), which contribute to
the development of heart disease.
5. Anti-ageing properties: Guavas are rich in vitamin A, vitamin C and antioxidants
like carotene and lycopene which help protect the skin from wrinkles. A guava a
day, keeps fine lines away.
6. Good for our brain: Guavas contain vitamin B3 and vitamin B6, also known as
niacin and pyridoxine respectively, which help in improving blood circulation to
the brain, stimulating cognitive function and relaxing the nerves.
Thus there are several aspects in which guava is helpful in maintaining our physical and
mental health.

What is Oxalate?
Oxalate is a naturally occurring organic (carboxylic) acid, primarily found in abundance
in plants, animals and humans. In plants, oxalate helps to get rid of extra calcium by
binding with it. That is why so many high-oxalate foods are derived from plants. Our
body either produces oxalate on its own or it converts other molecules like vitamin C to
oxalate. Foods that are high in vitamin C can increase the body’s oxalate levels. Vitamin

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C converts to oxalate. Levels over 1,000 milligrams (mg) per day have been shown to
increase oxalate levels.

Fig. 3: Structure of an oxalate molecule

External sources like the foods we eat also contribute to the accumulation of oxalate
in our body. When we eat foods with oxalate, it travels through the digestive tract and
passes out in the stool or urine. As it passes through the intestines, oxalate can bind with
calcium and be excreted in the stool.
Taking antibiotics, or having a history of digestive disease, can also increase the
body’s oxalate levels. The good bacteria in the gut help get rid of oxalate, and when the
levels of these bacteria are low, higher amounts of oxalate can be absorbed in the body.

Sources of Oxalate
Foods that are highest in oxalate include
 fruits
 vegetables
 nuts
 seeds
 legumes
 grains
High-oxalate fruits include
 berries
 kiwis
 figs
 purple grapes

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Vegetables that contain high levels of oxalate include
 rhubarb
 okra
 leeks
 spinach
 beets
 Swiss chard
Some grain products are also high in oxalate, including
 bran flakes
 wheat germ
 quinoa
The following foods are also high in oxalates
 cocoa
 chocolate
 tea

Oxalate: Good or Bad?

However, oxalate is not an essential molecule and is excreted from our body in an
unchanged form. Once consumed, oxalate can bind to minerals to form compounds,
including calcium oxalate and iron oxalate. This mostly occurs in the colon, but can
also take place in the kidneys and other parts of the urinary tract. For most people, these
compounds present in the body are then eliminated in the stool or is excreted in our
urine as a waste. When too much oxalate continues through to the kidneys, it results in
a medical condition called as hyperoxaluria, commonly referred to as kidney stones.
The higher your levels of oxalate, the greater your risk of developing these kinds of
kidney stones. Accordingly, diet is looked upon as a preventive measure in addition to
medicines to treat kidney stones.
Another major health concern about oxalate is that it can bind to minerals in the gut
and prevent the body from absorbing them. For example, spinach is high in calcium and
oxalate, which prevents a lot of the calcium from being absorbed into the body.
Eating fiber and oxalate together may further hinder nutrient absorption. Nevertheless,
it is important to remember that only some of the minerals in our food will bind to
oxalate. Even though calcium absorption from spinach is reduced, calcium absorption
from milk is not affected when milk and spinach are consumed together.

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 Some proponents of low-oxalate diets say people are better off not consuming foods
rich in oxalates, since they may have negative health effects. However, it is not that
simple. Many of these are healthy foods that contain important antioxidants, fiber and
other nutrients. Therefore, it is not a good idea for most people to completely stop eating
high-oxalate foods.

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 Requirements
Apparatus Required
1. 100 ml measuring flask
2. Pestle and mortar
3. Titration flask
4. 250 ml Beaker
5. Funnel
6. Burette
7. Weighing machine
8. Pipette

Fig. 4: Measuring flask Fig. 5: Titration Flask

Fig. 6: Beaker Fig. 7: Weighing machine Fig. 8: Pestle and mortar

Fig. 9: Burette and pipette Fig. 10: Funnel

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Chemicals Required
1. Dilute sulphuric acid (H2SO4)
2. 0.05N Potassium Permanganate (KMnO4) solution

Fig. 11: Chemicals required Fig. 12: Potassium Permanganate

Other Materials Required

1. Filter paper
2. Guava fruit at different stages of ripening

a.

Fig. 13: Filter paper Fig. 14: Guava at different

stages of ripening

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 Theory
Oxalate ions are extracted from the fruit by boiling pulp with dilute sulphuric acid. Then
oxalate ions are estimated volumetrically by titrating the solution with standard
Potassium Permanganate solution.

Titration is a common laboratory method of quantitative chemical analysis that is

used to determine the unknown concentration of a known reactant. Because volume
measurements play a key role in titration, it is also known as volumetric analysis.
A reagent, called the titrant or titrator, of a known concentration (a standard solution)
and volume is used to react with a solution of the analyte or titrand, whose
concentration is not known. Using a calibrated burette or chemistry pipetting syringe to
add the titrant, it is possible to determine the exact amount that has been consumed
when the endpoint is reached. The endpoint is the point at which the titration is
complete, as determined by an indicator. This is ideally the same volume as
the equivalence point — the volume of added titrant at which the number of moles of
titrant is equal to the number of moles of analyte, or some multiple thereof (as
in polyprotic acids). In the classic strong acid-strong base titration, the endpoint of a
titration is the point at which the pH of the reactant is just about equal to 7, and often
when the solution takes on a persisting solid colour as in the pink of
phenolphthalein indicator.

Fig. 15: Indicators used in titration

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Molecular Reaction Equations
2KMnO4 + 3H2SO4 K2SO4 + 2MnSO4 + 2H2O + 4[O]

60 70o C
HOOC-COOH.2H2O + [O]  
 2CO2 + 2H2O × 5

3KMnO4 + 3H2SO4 +5 HOOC-COOH.2H2O

K2SO4 + 2MnSO4 + 18H2O + 10CO2

Ionic Reaction Equations

MnO4- + 16H+ + 5e- Mn2+ + 4H2O × 2

C2O4 2CO2 + 2e- × 5

2MnO4- + 16H+ + 5C2O42- 2Mn2+ + 8H2O + 10CO2

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 Experimental Procedure
1. Weigh 50 g of fresh guava and crush it to a fine pulp using pestle-mortar.

2. Transfer the crushed pulp to a beaker and add about 50 ml dilute sulphuric acid to
it.

Fig. 16: Step 1 Fig. 17: Step 2

3. Boil the contents for about 10 minutes.

4. Cool and filter the contents in a 100 ml measuring flask. Make the volume up to
100 ml by adding distilled water.

Fig. 18: Step 3 Fig. 19: Step 4

5. Take 20 ml of the solution from the measuring flask into a titrating flask and add
20 ml of dilute sulphuric acid to it. Heat the mixture to about 60ᵒC and titrate it
against 0.05N Potassium Permangante solution taken in a burette. The end point is
appearance of permanent light-pink color.

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 Fig. 20: Step 5 – Titration

6. Repeat the above experiment with 50 g of 1, 2 and 3 days old guava fruit.

Precautions
1. Potassium permanganate solution is always taken in the burette.
2. Avoid the use of burette having a rubber tap as KMnO4 attacks rubber.
3. In order to get some idea about the temperature of the solution touch the flask to
the back side of your hand. When it becomes unbearable to touch, the required
temperature is reached.
4. Add about an equal volume of dil. H2SO4 to the guava extract to be titrated (say a
full test tube) before adding KMnO4 .
5. Read the upper meniscus while taking burette reading with KMnO4 solution.
6. In case, on addition of KMnO4 a brown ppt. appears, this shows that either H2SO4
has not been added or has been added in insufficient amount. In such a case, throw
away the solution and titrate again.
7. The concentration of oxalate ion obtained from the filtrate is unfavourably high. If
the titration is carried with concentrated filtrate, the scale of the burette is out of
scope in regard to the concentration of the oxalate ion. So diluting the concentrated
filtrate to an appropriate proportion and titrating it with KMnO 4 and estimating the
end point will make it favourable. After estimating the end from the diluted
solution, the original concentration of the concentrated filtrate can be calculated by
multiplying the concentration the estimated filtrate with the factor by which it was
diluted.

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 Observations
 Weight of guava fruit taken each time = 50.0 g
 Volume of guava extract taken in each titration = 20.0 ml
1
 Normality of KMnO4 solution = N
20

Burette readings Concordant volume

Guava Strength of
of the N/20 KMnO4
extract from Initial Final oxalate
solution used (ml)

Fresh guava

One day old

guava
Two days
old guava
Three days
old guava

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 Calculations and Results
 Let, N1 and N2 be the normality of the guava extract solution and the KMnO4
solution, respectively.
 Let, V1 and V2 be the volume of the guava extract and the KMnO 4 solution,
respectively, corresponding to the equivalence point in the titration process.
 Then, we have N1 × V1 = N2 × V2
 For our experiment, given V1 = 20 ml, N2 = 1/20 and observed V2 = x (say)
 Therefore, we calculate Normality of oxalate as N1 = x ÷ 400
 Strength of oxalate in guava extract = Normality × Equivalent mass of oxalate ion
= x / 400 × 44 g/litre of diluted extract

Experimental Results

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 Conclusion
This project aimed at estimating the amount of oxalate present in the guava fruits and
how the strength of oxalate present in the guava changes during the different stages of
ripening. Guava has the highest percentage of vitamin C among all the citrus fruits. It
also contains oxalate amount of which varies with ripening of the fruit. Oxalate form an
insoluble complex with calcium in the urine resulting in kidney stone, a condition
medically known as hyperoxaluria. Therefore, presence of oxalate in our food is
injurious to health and hence, oxalate rich foods are usually restricted to some degree,
particularly in patients with high urinary oxalate level.

In our experiment, we found that the normality of oxalate ions in fresh guava
solution is 0.0875 N, 0.0925 N and 0.0975 N in semi-ripened guava (one and two days
old guava, respectively), while it is 0.1 N in 3-days old fully ripened guava. The
respective strength of the oxalate ions in guava extract solution was found to be 3.85,
4.07, 4.29 and 4.4 gram per litre, averaging at 4.1525 gram per litre during ripening of
guava over four days.

On the basis of our experimental observations and results obtained, it is therefore

concluded that –

 Presence of oxalate ion is very high in guava.

 The strength of oxalate ions increases as the guava fruit ripens. The oxalate
content was on the increase in the fruit as the days passed on, that is as the
ripening proceeded. During ripening of guava, the oxalate content increases
progressively and the fully ripe fruit has the maximum oxalate content.

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 Bibliography
[1] N.K. Verma, B.K. Vermani, N. Verma, K.K. Rehani, Comprehensive Practical
Chemistry, Laxmi Publications, New Delhi.
[2] O. Traxer, B. Huet, J. Poindexter, C.Y. Pak and M.S. Pearle, “Effect of ascorbic
acid consumption on urinary stone risk factors,” Journal of Urology, vol. 170,
no. 2, pp. 397-401, Aug. 2003.
[3] S.C. Noonan and G.P. Savage, “Oxalate content of foods and its effect on
humans,” Asia Pacific Journal of Clinical Nutrition, vol. 8, no. 1, pp. 64-74, Mar.
1999.
[4] R.P. Heaney and C.M. Weaver, “Oxalate: effect on calcium absorbability,”
American Journal of Clinical Nutrition, vol. 50, no. 4, pp. 830-832, Oct. 1989.
[5] https://www.healthline.com/nutrition/oxalate-good-or-bad#section1
[6] https://www.healthline.com › health › what-is-a-low-oxalate-diet

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