1. DEPARTMENT OF BIOLOGY
FACULTY OF SCIENCE AND MATHEMATICS
UNIVERSITI PENDIDIKAN SULTAN IDRIS
TBF 3023
PLANT PHYSIOLOGY
MINI PROJECT:
FRUIT SEEDS COMPOSITION (BIOCHEMICAL ANALYSIS)
GROUP MEMBERS : CHEW MEI PING D20091034816
ONG SHWU CHYN D20091034817
PUI WAN TING D20091034840
GROUP : A
LECTURER : DR. HASIMAH ALIMON
SUBMITTION DATE : 20 OCTOBER 2010

2. Fruit Seeds Composition (Biochemical Analysis)
M.P. CHEW, S.C. ONG AND W.T. PUI
Department of Biology,
Faculty of Science and Mathematics,
Universiti Pendidikan Sultan Idris,
35900 Tanjung Malim, Perak, Malaysia.
ABSTRACT
The seed of six types of fruits which is Artocarpus heterophyllus (jackfruit), Artocarpus
integer (cempedak), Nephelium lappaceum (rambutan), Dimocarpus longan (longan),
Lansium domesticum (duku), and Annona squamosa (nona) were examined in term of seed
biochemical composition. These six types of seed fruit are full of their own nutrient. In this
project, all these seed of fruit is tested for carbohydrate, protein and lipid composition.
Carbohydrate and protein are present in all the types of seed while none of them showing the
present of lipid.
Keywords: Artocarpus integer, Artocarpus heterophyllus, Nephelium lappaceum, Lansium
domesticum, Annonaceae squamosa, Dimocarpus longan, seed biochemical composition,
carbohydrate, protein, lipid.
INTRODUCTION
Artocarpus heterophyllus, Artocarpus integer, Nephelium lappaceum, Dimocarpus
longan, Lansium domesticum, and Annona squamosa are seasonal local fruit which can be
found in Malaysia. All of them are flowering plant (Angiosperm). The Artocarpus integer
and Artocarpus heterophyllus are from Moraceae family, which often called the mulberry
family, comprising about 40 genera and over 1000 species. The generic name of Artocarpus
integer comes from the Greek words „artos‟ (bread) and „karpos‟ (fruit); the fruits are eaten
and are commonly called breadfruit. Common name for Artocarpus integer is cempedak.
Artocarpus integer is native to Southeast Asia and exotic in Indonesia, Malaysia, Myanmar,
Thailand and Vietnam. Its fruit is sausage-shaped range from 22 to 50 centimetres in length
and 10 to 17 cm across. Each fruit of Artocarpus integer contains many kidney-shaped seeds
with a thin and white coriacous testa. Artocarpus integer seeds are rich in starch and are eaten.
Common name for Artocarpus heterophyllus is jackfruit (English) or nangka (Malay). The
name of „jackfruit‟ is derived from the Portuguese, Jaca. Artocarpus heterophyllus has a
compound or multiple fruit syncarp. Its fruit is oblong-cylindric in shape, typically 30-40 cm

3. in length and 4.5-30 kg in weight. The seeds of Artocarpus heterophyllus are light brown to
brown, rounded in shape, 2-3 cm in length, 0.4-0.6 in diameter and enclosed a thin whitish
membrane. We can found up to 500 seeds in each fruit. Their seeds can be stored up to a
month in cool and humid condition. The seeds also can be boiled or baked like beans before
eaten.
The Dimocarpus longan and Nephelium lappaceum are from Sapindaceae family,
also known as the soapberry family. The common name for Dimocarpus longan are longan
(English), longyan, dragon eye (Vietnam), mata kucing (Malay). It is called dragon eye‟ as
this its literal translation from Hokkien (Amoy). When the fruit is shelled, it looks like an
eyeball as the black seed shows through the translucent flesh like a pupil. It is also known as
the “little brother of the lychee” or “slave of the lychee”, as the lychee is more glamorous
than the longan. Dimocarpus longan is native to Southeast Asia. Their fruits are round in
shape, 1-2.5 cm in diameter with a rough light brown peel and sweet translucent, whitish pulp
surrounding a smooth black seed. The common name of Nephelium lappaceum is rambutan
(Malay). Nephelium lappaceum is originally from Malaysia and Indonesia but now cultivated
throughout the tropics. The name rambutan is derived from the Malay word, rambut which
mean hairy because the appearance of its fruit, the hair covers the surface of the fruit. The
seed of Nephelium lappaceum resemble an almond and each fruit only contain single seed.
Then, Lansium domesticum is come from Meliaceae family or the Mahogany family
and mostly trees and shrubs (and a few herbaceous plants). The common names of Lansium
domesticum are duku and langsat. Lansium domesticum is native in Thailand, Malaysia and
Indonesia. Their fruits are spheroid, ellipsoid or ovoid, around 2.5-5cm in diameter and
usually in clusters of 4-40. Each fruits usually contain 1-3 greenish seeds. The seeds are
relatively 2-2.5 cm long and 1.25-2 cm wide, very bitter and sometimes if the flesh clings
tightly to the seed; it may acquire some of its bitterness.
Next, Annonaceae squamosa is come from Annonaceae family also called the custard
apple family. The common name of Annonaceae squamosa is sugar apple and custard apple
(English), buah nona (Malay). Its fruit is round or heart-shaped, 5-10 cm in diameter and
form of loosely cohering or almost free carpels. Each carpel contains an oblong, shinny and
smooth black seed which are 0.5-1.0cm long. These seeds cannot be eaten because it is
poisonous and contain highly toxic substances.
All the seeds of fruit are rich in nutrient and contain some of the biochemical
composition. Carbohydrate and protein are found in all fruits seeds that been examined. The
lipid is not found in any seeds of fruits.
MATERIALS AND METHODS
Seeds from fruit of 4 families had been used for this project. The fruit seeds used were
Artocarpus heterophyllus (jackfruit), Artocarpus integer (cempedak), Dimocarpus longan
(longan), Nephelium lappaceum (rambutan), Lansium domesticum (duku) and Annona

4. squamosa (nona). The pulps and fleshes of each fruit were removed and the seeds were taken
to test its biochemical composition.
There are few chemical reagents had been used for test the biochemical composition
in the seeds such as 5% KOH solution, 1% Copper sulphate solution, Millon solution, Alfa
naftol 5% solution, concentrated sulphuric acid, Barfoed solution, Iodine solution, Benedict‟s
reagent, Sudan IV solution and distilled water. The apparatus that are required for the
experiment are stirring hotplate for boil the solution, beakers, test tubes and test tube shelf to
collect the seed solutions, mortar and pestles to grind the fruit seeds.
Each of the fruit seeds was ground with some distilled water in a clean mortar. For
those seeds especially Artocarpus sp. are hard to grind like other seeds. Thus, we cut them
into small pieces before grind it. Then, each of the seed solutions were collected into different
clean test tubes. Each of the seed solutions was tested for carbohydrate, protein and lipid
composition.
Molisch test, Barfoed test, Benedict‟s test and iodine test were used to test the
carbohydrate composition in each type of fruit seeds. In Molisch test, 2ml of each seed
solution were filled into 6 separated clean test tubes respectively. 5 drops of Alfa naftol 5%

5. were added into each test tube respectively and the mixtures were shook. After that, 1ml of
concentrated sulphuric acid was slowly filled into the 7 test tube and did not shake the test
tubes. The changing in colour of the mixtures was observed and recorded. For the Barfoed
test, 6 test tubes were filled with 5ml Barfoed solution and then 2ml of each seed solutions
were added to the test tubes respectively. The colour appearance of the mixtures was
observed and recorded. For Benedict‟s test, 2ml of each type of seed solutions were filled
into 6 separated clean test tubes and 2ml of Benedict‟s reagent were added into it. After that,
the mixtures were boiled for 1 minute by using stirring hotplate. The changing of colour in
each mixture was observed after it had cooling down. Few drops of iodine were added into
the test tubes with each type of seed solution for iodine test. The results of colour change in
the mixtures were observed.
There are two tests to indicate the presence of protein in the fruit seeds composition
which are Biuret test and Millon test. For Biuret test, 2ml of each type of seed solution were
filled into 7 separated test tubes respectively. 2ml of 5% potassium hydroxide (KOH) were
added to each solution and mixed completely. Then, 2 drops of 1% copper sulphate were
added to the mixtures. The results of colour change in mixtures were observed. While for
Millon test, 2ml of each type of seed solutions were filled into 7 separated test tubes and 3
drops of Millon solution were added into the solutions. The changing of colour in mixtures
was observed.
Colouring test was used to test the presence of lipid in fruit seeds. 2ml of each type of
seed solutions were filled into 6 separated clean test tubes. Then, 3 drops of Sudan IV
solution were added to the seed solutions respectively. Then, 5 ml of distilled water also had
been poured into each mixture later. The mixtures were shaking. The results of colour change
in mixtures were observed.
RESULTS AND DISCUSSION
The purpose of Molisch test is to indicate the presence of carbohydrate by observing purple
ring form at the interface between concentrated sulphuric acid and solution. The results
showing the data for six different types of fruit seeds after Molisch test are given in Table 1.
Table 1- Results of fruit seeds composition in Molisch test
Fruit seeds Colour change
Actocarpus heterophyllus Black precipitate, purple ring on the surface of solution
Actocarpus integer Black precipitate, purple ring on the surface of solution
Nephelium lappaceum Black precipitate, purple ring on the surface of solution
Dimocarpus longan Black precipitate, purple ring on the surface of solution
Lansium domesticum Black precipitate, purple ring on the surface of solution
Annona squamosa Black precipitate, purple ring on the surface of solution

6. Artocarpus heterophyllus, Artocarpus integer, Nephelium lappaceum, Dimocarpus longan,
Lansium domesticum, and Annona squamosa contain carbohydrate not only in their flesh but
also in their seeds. A layer of purple ring was shown in all test tubes as we can indicate the
presence of carbohydrate in all types of fruit seeds. The black precipitate found at the bottom
of solution is alpha naftol and it is not water soluble. Thus, it doesn‟t dissolve in those
solutions. It also because of carbohydrates undergoes dehydration reactions (loss of water) in
the presence of concentrated sulphuric acid. Pentoses and hexoses form five member oxygen
containing rings on dehydration. The five member ring, known as furfural, further reacts with
alpha-naphthol 5% to form colored compounds. Purple colour ring is form when alpha-
naphthol 5% reacts with the cyclic aldehydes.
In Barfoed test, the presence of monosaccharide is indicating by observed the
formation of brick-red precipitate of copper (II) oxide in the solution. The results are shown
in Table 2.
Table 2- Results of fruit seeds composition in Barfoed test
Fruit seeds Colour change
Actocarpus heterophyllus Light blue. No changes of colour.
Actocarpus integer Light blue to dark blue
Nephelium lappaceum Light Blue to blue green, white ring on the surface of
solution
Dimocarpus longan Light Blue to dark blue, red precipitate, red ring on the
surface of solution
Lansium domesticum Light Blue to blue, white ring on the surface of solution
Annona squamosa Light blue. No changes of colour.

7. In Artocarpus heterophyllus, Artocarpus integer and Annona squamosa, there hasn‟t any
colour changes in the solution and it remain blue. This means, these fruit seeds do not
composed monosaccharide. Although Nephelium lappaceum and Lansium domesticum show
white ring on the surface of solution, but this couldn‟t indicate that the seeds contain
monosaccharide. Only Dimorcarpus longan shows red precipitate in its solution. This means,
monosaccharide can be found in its seeds.
Table 3- Results of fruit seeds composition in Benedict‟s test
Fruit seeds Colour change
Actocarpus heterophyllus No change of colour
Actocarpus integer Light blue to dark blue
Nephelium lappaceum Light blue to light green after mix, but doesn‟t change after heat.
Dimocarpus longan Light blue to light green, red precipitate
Lansium domesticum Light blue to orange, brick red precipitate
Annona squamosa No change of colour
For Benedict‟s test, reducing sugar is indicated
presence if blue colouration of mixture turns green,
yellow, orange and form a brick-red precipitate. Non
reducing sugar will not show any effect in colour
change. The results are shown in Table 3.

8. Seed of Actocarpus heterophyllus, Actocarpus integer, and Annona squamosa show negative
result in Benedict‟s test as they have no reducing sugar. For Nephelium lappaceum, it also
doesn‟t contain redusing sugar in its seed because the solution changes to green colour as
soon as it mixes with Benedict‟s reagent. It doesn‟t show any changes after heat. Reducing
sugar is present in Dimorcarpus longan and Lansium domesticum as the brick red precipitate
is clearly shown in the solution. Benedict Solution is light blue because it contains copper
sulphate. When it is mixed and heated with a sugar, such as glucose, which has electrons
available to donate, the copper will accept the electrons and become reduced, which turns it
brownish-orange. During this process, the blue copper (II) ion is reduced to a red copper (I)
ion. While the copper is being reduced, the glucose gives up an electron and is oxidized.
Because glucose is able to reduce the copper in Benedict Solution, we call it a reducing sugar.
To indicate the presence of starch, iodine test has been carried out. The solutions were
test by drop a few drops of iodine in it and observed the colour change. For those seed which
is harder to smash, we cut them into small pieces and drop the iodine on it and observed the
colour change. The results are shown in Table 4.
Table 4- Results of fruit seeds composition in Iodine test
Fruit seeds Colour change
Actocarpus heterophyllus Dark purple
Actocarpus integer Dark purple
Nephelium lappaceum Black precipitate at the bottom of test tube (many)
Dimocarpus longan Black precipitate at the bottom of test tube
Lansium domesticum Black precipitate at the bottom of test tube (less)
Annona squamosa Black precipitate at the bottom of test tube

9. All of the fruit seeds are fairly rich in starch. These fruit seed solutions had shown dark
purple colour when few drops iodine are added. This reaction is the result of the formation of
polyiodide chains from the reaction of starch and iodine.
Biuret test has carried out to test the presence of protein in fruit seeds. Purple or blue
colouration form in the solution indicates the presence of protein. There is no heating
required in this test. Table 5 is showing the results in this test.
Table 5- Results of fruit seeds composition in Biuret test
Fruit seeds Colour change
Actocarpus heterophyllus White solution, with light blue ring appear on the surface
Actocarpus integer White solution, with light blue ring appear on the surface
Nephelium lappaceum Yellowish solution, with light blue ring appear on the surface
Dimocarpus longan Brownish solution, with dark blue ring appear on the surface
Lansium domesticum Brownish solution, with dark blue ring appear on the surface
Annona squamosa White solution, with blue ring appear on the surface

10. Blue ring is appearing on the surface of each solution. This means protein is present in all six
types of fruit seeds. In solution of Dimocarpus longan and Lansium domesticum, dark blue
ring is appear. This is because their seed contain higher amount of protein and this make the
seeds taste bitter.
For Millon‟s test, it also used to indicate the presence of protein. The presence of
white precipitate in the solution shows that the seeds contain protein. It will probably
coagulate on heating and turns red. But, Millon‟s reagent is poisonous, so heating is not
encouraging.
Table 6- Results of fruit seeds composition in Millon‟s test
Fruit seeds Colour change
Actocarpus heterophyllus White precipitate appear
Actocarpus integer White precipitate appear
Nephelium lappaceum White precipitate appear
Dimocarpus longan White precipitate appear
Lansium domesticum White precipitate appear
Annona squamosa White precipitate appear
To test whether lipid is present in these six different types of seeds, colouring test is
used. If there is lipid contain in the seed, a red-stained layer will separates on the surface of
water. Results are shown in Table 7 below.
All of the fruit seeds give
positive results in Millon‟s
test by showing white
precipitate in the solution.
There are more precipitate
appear in Annona
squamosa solution than in
other seed solution because
Annona squamosa has
higher concentration.

11. Table 7- Results of fruit seeds composition in colouring test.
Fruit seeds Colour change
Actocarpus heterophyllus No change
Actocarpus integer No change
Nephelium lappaceum No change
Dimocarpus longan No change
Lansium domesticum Black precipitated at bottom of test tube
Annona squamosa Black precipitated at bottom of test tube
All of the fruit seeds solutions remain unchanged in this test because they do not contain lipid
in it. The black precipitate in the bottom of each test tube is because of the Sudan IV which
had added in it.
Some electronic journal has been read through to get some brief idea about fruit seed
composition. From all the journal which have read, some importance and significant uses can
be point out and it will be useful for public.
The seeds of Artocarpus heterophyllus are an excellent addition to curries or can be
eaten freshly, cooked or dried with salt as a snack. The cooked and dried seeds are milled to a
flour-like consistency and added to bread dough. (Craig R. Elevitch and Harley I. Manner,
Artocarpus heterophyllus) The seed starch is useful in relieving biliousness while the roasted
seeds are regarded as aphrodisiac. In China, the seeds of Artocarpus heterophyllus are
consider as cooling and nitrous tonic. On the other hand, the seeds of Artocarpus integer are
rich in starch and can be eaten after cook or boil.
While in our analysis from journal of M.A. Austin and B.C. Chua (1988), it shows the
seeds of Nephelium lappaceum contained some protein. The seed is also soft and crunchy.
They are mildly poisonous when raw but may be cooked and eaten.
Crushed seed of Dimocarpus longan produce foam, which is used as shampoo like
soapberries (Sapindus saponaria L.), due to its saponin content according to Wong Kai Choo,
2000.

12. For the powdered seeds of Lansium dom 1esticum, they are used to reduce fever.
Besides that, grounded seeds mixed with water as vermifuge and antipyretic (Philippine
Medical Plants). While the powdered seeds of Annona squamosa are used to kills fleas and as
a parasiticide and insecticide especially against body and head-lice. A high potency is
ascribed to the presence of a glyceride of a hydroxylated unsaturated acid. (Burkill, H.M.
1985. The Useful Plants of West Tropical Africa, Vol 1).
CONCLUSION
This mini project helps us to get more information about the biochemical composition in
different type of local fruit and it is easy to conduct even in secondary educational level.
Artocarpus heterophyllus, Artocarpus integer, Nephelium lappaceum, Dimocarpus longan,
Lansium domesticum, and Annona squamosa are test for biochemical composition;
carbohydrate, protein and lipid. All type of seeds show positive result for carbohydrate and
protein test but do not show response for lipid test.

13. APPENDIX
REFERENCES
"Barfoed's test." A Dictionary of Biology. 2004. Retrieved October 19, 2010 from
Encyclopedia.com: http://www.encyclopedia.com/doc/1O6-Barfoedstest.html
Berg, L. (2008). Introductory Botany: Plants, people and the Environment. 2nd.
Ed.:
Thomson Brooks/Cole Publishing Company. USA.
Brooke Yool (2010). The Effect of Benedict Solution on Glucose. Obtained on June 3,
2010 from
http://www.ehow.com/about_6584528_effect-benedict-solution-glucose.html
Craig R. Elevitch and Harley I. Manner (2006). Artocarpus heterophyllus (jackfruit).
Obtained on April 2006 from
http://www.agroforestry.net/tti/A.heterophyllus-jackfruit.pdf.
Lee Ching and . Arunasalam (2007). Longman Pre-U Text STPM Biology Volume 2.
Selangor: Pearson Malaysia Sdn. Bhd.
M.A. Augustin and B.C. Chua (1988). Composition of Rambutan Seeds. Obtained on
July 21, 1987 from
http://psasir.upm.edu.my/2559/1/Composition_of_Rambutan_Seeds.pdf.
Orwa C, Mutua A , Kindt R , Jamnadass R, Simons A. 2009. Agroforestree
Database:a tree reference and selection guide version 4.0
http://www.worldagroforestry.org/af/treedb/
Oxford (2004). Dictionary of Biology (fifth edition). New York: Oxford University
Press.
Philippine Medical Plants (2010). Lansones. Obtained on June 2010 from
http://www.stuartxchange.org/Lansones.html.
Rost, Barbour, Stocking, Murphy (2006). Plant Biology, Second Edition. New York:
Thomson Higher Education.
Stern, K.R. (2006). Introductory Plant Biology 10th.
Ed.: Mc Graw Hill. USA.
Wong, Kai Choo (2000). Longan Production in Asia. Obtained on December 2000
from http://www.fao.org/DOCREP/003/X6908E/X6908e00.htm#Contents.