PHYTOCHEMICAL SCREENING OF SOURSOP SEED (Annona
muricata)
ABSTRACT
They phytochemical screening of ethanolic extract of Annona muricata seeds
was carried out using standard method. The phytochemical compounds
screened were Saponins, flavonoids, Tannins, Alkaloids and cardiac glycosides.
The result shows that saponins is moderately present, flavonoids and Alkaloids
is Abundantly present, Tannins is Trace present and test for cardiac glycosides,
Liebermans and salkowskis test is trace present. Based on the experiment carry
out the result shows that Annona muricata seeds can be used for medicinal
activity.
Key Words: Annona muricata, flavonoids, Alkaloids, Tannins, Ethanolic
Screening.
TABLE OF CONTENTS
Title Page
Certification
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Dedication ...
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Acknowledgements
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Abstract
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Table of Contents ...
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vii-vix
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List of Tables
CHAPTER ONE
1.0
Introduction
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1.1
Background of the Study ...
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1-2
1.2
Aim and Objectives
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1.3
Scope and Limitation
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1.4
Definition of Terms
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CHAPTER TWO
2.0
Literature Review
2.1
Botanical Classification of Soursop
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2.2
Description of Soursop
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4-5
2.3
Distribution of Soursop ...
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5-6
2.4
Pharmacological Activities of Soursop
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2.4.1 Anti-Arithrific Activity ...
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2.4.2 Anti-Cancer Activity
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2.4.5 Anti-Inflammatory Activity
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2.4.6 Anti-Oxidant Activity
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2.4.7 Anti-Parasitic Activity
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13-14
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2.4.3 Anti-Convulsant Activity
2.4.4 Anti-Diabetic Activity
2.5
Phytochemical
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2.6
Major Phytochemical Found in Plant ...
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2.6.1 Alkaloids
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14-15
2.6.2 Flavonoids ...
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2.6.3 Saponnins ...
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2.6.4 Tannins
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2.6.5 Phenol
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CHAPTER THREE
3.0
Materials and Method
3.1
Materials and Reagents Used
3.2
Sample Collection
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3.3
Sample Preparation
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3.4
Phytochemical Screening
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3.5
Method of Phytochemical Screening ...
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3.5.1 Test for Saponins ...
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3.5.2 Test for Flavonoids
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3.5.3 Test for Tannins
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3.5.4 Test for Alkaloids ...
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3.5.5 Test for Cardiac Glycoside ...
CHAPTER FOUR
4.0
Result and Discussion
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4.1
Result
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Discussion ...
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CHAPTER FIVE
5.0
Summary, Conclusion and Recommendation
5.1
Summary
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Conclusion
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Recommendation ...
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References
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CHAPTER ONE
1.1
Background of Study
Soursop (Annona muricata) is a peculiar fruit with an inedible skin covered
with many soft spines. It is an evergreen, terrestrial, erect tree reaching 5-8m in
height and features and open, roundish canopy with large, glossy, darkgreen
leaves. The edible fruits of the tree are large, heart shaped and green in colour
and the diameter varies between 15 and 20cm. soursop is native to central
America, the Caribbean and northern south America, Columbia and Brazil,
Mexico, peru and Venezuela and some sub-saharan African countries that is
within the tropics (Morton, 2013). The unripe fruit is green, hard both on the
inside and outside and pulp is white. Soursop has a combination of strawberry
and pineapple with sour citrus flavor notes, contrasting with an underlying
creamy flavor reminiscent of coconut and banana (Abdi 2011). The fruit is used
as natural medicine for arthritic pain, nevralgia, arthritis, diarrhea, fever,
malaria, parasites, rheumatism, skin rashes and worms. And it also elevate a
mother’s milk production after child birth. The leaves are employed to treat
cystitis, diabetes, headaches and insomnia. The crushed seeds are believed to
have anthelimintic activities against external and internal worms and parasites.
The fruit and flowers are employed as remedies against catarrh, while the root
bark and leaves are believed to have antiphlogistic and antheklmintic activities
(Mata et al., 2005). Food and Agriculture Organization (FAO) data show that
global commercialization of fruit derived products has grown significantly and a
reduction in consumption of ready-to-drink fruit based beverages has been
observed (Zulucta et al., 2007).
1.2
Statement of the problem
The study was limited by time, lack of adequate facilities and financial
constraint.
1.3
Aim and Objectives
The Aim and objectives of the project is to do a comparative analysis on the
phytochemicals present in the sour Sop seed (Annona muricata).
1.4
Significance of the Study
The importance of this study includes:
 To understand the pharmacological functions of annona muricata
 To determine selected nutritional and phytochemical properties of the
leaves of annona muricata
1.5
Scope of the study
The design of this research projects is to investigate and determine the
phytochemical composition of Sour Sop seed (Annona muricata).
1.6
Definition of Terms
PHYTOCHEMICALS:
Phytochemicals are chemical compounds produced by plants, generally to help
them thrive or thwart competitors, predators, or pathogens. The name comes
from the Greek work phyton, meaning plant. Some phytochemicals have been
used as poisons and others as traditional medicines.
ANNONA MURICATA:
Annona Muricata is a species of the genus Annona of the Custard apple tree
family, Annonaceae, which has edible fruit. The fruit is usually called sour sop
due to it’s slightly acidic taste when ripe. Annona muricata is a native of the
caribean and central America but is now widely cultivated.
CHAPTER TWO
LITERATURE REVIEW
2.1
2.2
Botanical Classification of Sour Sop
Kingdom
Plantae
Subkingdom
Tracheobionata
Super division
Spermatophyte
Division
Magnoliophyta
Class
Magnoliopsida
Subclass
Magnoliidae
Order
Magnoliales
Family
Annonaceae
Genus
Annona L.
Species
Annona muricata L. (Morton 2013)
Description of Sour Sop
The sour sop from the Annonaceae family is found to be the most important
tropical fruit that contributes much to the wider economic growth in some
tropical countries such as tropical America, African and Malaysia (Bentl;ey et
al., 1998). Is a small, upright, evergreen tree that can grow to about Four (4)
metres (13 ft) tall. Its young branches are hairy. They are glossy dark green with
no hairs above and paler and minutey hairy to no hairs below. Leaves are
Oblong to oval 8 centimeters (3.1 in) to 16 centimeters (6.3in) long and 3
centimeters (1.2in) to 7 centimeters (2.8 in) wide. (Bentley et al., 1998). The
leaf stalks are 4 millimeters (O.16in) to 13 millimeters (0.5) in long and without
hairs. They appear opposite from the leaves or as an extra form near the leaf
stalk, each with one or two flowers, occasionally a third minutely hairy to
hairless and 15 millimetres (0.59in) to 20 millimeters (0.7) with small bracelets
nearer to the base which are densely hairy (parkh and chanda 2007). Sour sop
gives a flavor of custard when it is ripen condition and hence has a pleasant,
distinctive aroma and fibrous pulp that can be consumed because of its very
juicy, creamy and sweet character (Prasad m, et al., 2010).
2.3
Distribution of Sour Sop
Annona muricatais tolerant of poor soil and prefers lowland areas between the
actitudes of 0 metres (0ft) to 1,200 metres (3, 900 ft). it cannot stand frost, the
exact origin is unknown, it is native to the tropical regions of the Americas and
is widely propagated (Abraham Z. 1991).
2.4
Pharmacological activities of Sour Sop
2.4.1 Anti – arthritic Activity
A. muricta is among the ethnome diciries employed to treat arthritic pain. An in
vivo study on different doses (3,10,30 and 100mg/kg) of ethanolic extract from
A. muricata leaves has investigated the anti -0 arthritic activity in complete
freund’s adjuvant (CFA) Induced arthritis in rats. According to the result, oral
administration of the extract reduced the edema in a dose dependent manner
after two weeks of injection. The extract at higher dose was able to significantly
suppressed TNF- x and IL-IB expression in local tissues. The anti – arthritic
activity of A. muricata leaves contributed to the suppression of pro –
inflammatory cytokines (Chan and Artant. 2010). Hence, the anti – arthritic
potential of A. muricata was sub stantiated by the findings of this in vivo study.
2.4.2 Anti – Cancer Activity
An in vitro Studies were performed to determine the mechanism of action of
ethyl actate extract of A. muricata leaves against colon cancer cells (H T-29 and
HCT- 116) and lung cancer cells (A549). The leaf extract was able to induce
apoptosis in colon and lung cancer cells through the mitochondrial – mediated
pathway this anti proleferature effect was associated with celly cycle arrest in
the Gi. Phase (moghadamtous, et al. 2014). In addition the migration and
invasion of colon cancer cells were significantly inhibityed by the leaf extract.
The activation of caspase 3 by the ethanolic extract of the leaves also
demonstrated and apoptosis – inducing effect in myelogenous leukemic k562
cells, which was confirmed with a TUNEL assay (Ezirim et al., 2013). In Vitro
and in Vivo studies were performed on the water of the A. muricata leaves
against the benign prostatic hyperplasi (BPH -1) cell line of rats prostates. The
results showed a suppressive effect on BPH -1 cells with an IC50 value of 1.36
neg/ml after 72hours associated with an up – regulation of Bax and down –
regulation of BCl -2 at the MRNA level. After two months of treatment with the
extract (30 and 300mg/ml doses). The size of the rats prostates were decreased,
which was suggested to occur through apoptosis induction (AS are. Et al 2015).
This promising antitumor effect was reported in an in vivo study of 7, 12dimethyl benzene anthracene (DMBA) Induced cell proliferate on in the breast
tissues of mice. The protective effect against DNA induced by DMBA showed
that Oral administration of the A. muricata leaves extract, may have protective
effects against the development of breast carcinogenesis (minari et al 2014).
The leaves, even at the low dose of 30mg/ kg suppressed the initiation and
promotion stage of skin problem in mice that was induced by DMBA and croton
oil, respectively (Hamizah et al., 2012). Moghadamtousi and colleagues also
examined the in Vivo chemopreventive potential of the ethyl aetate extract of
the A. muricata leaves against azoxymethane – induced colonc aberrant
cryptfoci (ACF) in rats. The oral administration of the extract at two doses (250
and 500mg/kg) for
60 days, significantly reduced ACE blue staining of
colorectal specimens (moghadamtousi et al., 2014). The immune histochemistry
analysis showed that this activity was accompanied by the up – regulation of
BCL -2. This signiuficant reduction in ACF formation was also reported for the
ethanolic extract of the leave against 1,2- dimethyl hydrazine (DMH). Induced
colon cancer (Eggadi et al., 2014).
2.4.3 Anti – convulsant Activity
In African countries, the decoction of the A. muricata leaves is traditionally
used to control fever and convulsive seizures (Nghovemo et al., 1997). To
substantiate the anti convulsant activity of the leaves govemo and colleagues
investigated the effect of the ethanolic extract of the leaves pentylenetetrazol –
induced tonic colonic seizures in mice. The result showed that the plant extract
at 100 and 300 mig/kg doses significantly decreased the Incidence and the
mortality rate of tonic seizures. Administration of the extract to mice also
lengthened the onset of clonic seizures. This study showed that a subsequent
bioassay – guided investigation may lead to the isolation of a bioactive
compound that can be used as an anticovulsant drug (Govemo. et al., 1995).
2.4.4 Anti – diabetic Activity
The Chronic disease of diabetes milltus afflicts a large proportion of people all
around the world. Therefore, an effective natural adjuvant therapy would be
blindingly beneficial to diabetic complications and augment the quality of life of
diabetic patients. Due to the traditional application of A. muricata against
diabetes, several studies have investigated this potential in vivo. The extract at
the same dose significantly decreased the serum total cholesterol, low – density
lipopropein triglyceride and very low density lipoprotein (Adeyemi et al.,
2008). Based on the ethnopharmacological application of A. muricata leaves
against diabetes in Cameroon, another similar study examined the aqueous
extract of the leaves against strepto zotocin – induced diabetes in rats and
reported the same promising antidiabetic activities. This activity was explained
by it’s antioxidant and hypolipidemic potential and protective effects against
pancreatic B-cells (Florence et al., 2014). The stem bark ethanolic extract also
demonstrated promising anti diabetic increase blood glucose and was associated
with a reduction in cholesterol and triglyceride levels (Ahalya et al., 2014).
2.4.5 Anti – Inflammatory Activity
Oral treatment in rats with ethanolic leaf extracts of A. muricata (10,30,100 and
300mg/kg) significantly reduced carrageenan – Indued Edema in rat pawas by
79% in a dose – dependent manner, exhibiting it’s anti – inflammatory activities
(Roslida et al., 2014). This anti – inflammatory effect was accompanied by
reductions in the leukocyte migration and exudates volume (De Sousa et al.,
2010). Oral administration in mice with the same extract showed significant
suppression of abdominal contortions with acetic acid (0.69 o V/v). exhibiting a
powerful anti – nociceptive activity (Roslida et al., 2014). Inaddition the
formalin test and paw licking and hot plate responses also corroborated the
market analysis effect of the A. muricata leaves. (Hamid et al., 2010). The
protective effect of theA. muricata leaves against complete freund’s adjuvant
(CFA) – Induced arthritis in rats and xylem – Induced ear ederna in mice was
associated with an attenuation in the TNF – d and IL-IB protion expression,
demonstrating that the leaves could be used against both acute and chronic
inflammation. The same assay showed the anti – Inflammatory medicators and
interactions with the opioidergic pathway, respectively (Ishola et al., 2014).
2.4.6 Antioxidant Activity
Immoderate generation of Intracellular reactive oxygen species (ROS) is a
precursor of oxidative stress which subsequently catalyzes metabolic efficiency
and particular death. Through biochemical and physiological lesions (Chance et
al., 1999). The identification for antioxidants from natural in mean studies for
their noteworthy role in nullifying the destructive effects of ROS (Liao et al.,
2012). The antioxidant activity of the A. muricata leaves was found to be
stronger than A Squamosa and A reticulate species as shown through different
in Vitro models such as nitric oxide and hydroxyl radicals. (Baskar et al.,
2017). The seeds and leaves of the plant are reported to possess enzymatic
antioxidants, including catalase and superioxide dismutase and non – enzymatic
antioxidants, including Vitamin C and and E (Vijayameena et al.,
2013).
Padma and colleagues showed that the ethanolic extract of the A. muricata stem
bark caused a reduction in lipid peroxidation induced by cold immobilization
stress in the brain and liver of rats, indicating the adaptogenic potential of this
plant (Padma et al., 2013). The stem bark extract (200mg/kg) also showed
protective effects against oxidative stress induced by carbon tetrachloride in rats
and significantly increase the oxidant levels and serum enzyme activities to near
normal. (Olakunle et al., 2014).
2.4.7 Anti – Parasitic Activity
Protozoal infections cause debilitating diseases such as leishmaniasis and
trypanoso miasis, which have both afflicted a note worthy proportion of the
world population. The development of resistance to empirically discovered
drugs represents a major hindrance to treatment of protozoal diseases. Moreover
in case of long- term usage toxicity and several side effects have made the
available treatments more unsatisfactory. The ethlyl acetate leaf extract of A.
muricata was assuaged against three Leishmania species promising activity was
reported with values lower than 25ug/ml (Osorio et al., 2007). The same
promising antileishamanial effect was reported against L. braziliensis and L.
Panmensis species with a toxicity effect higher than Glucantine, which was used
as a positive control (Jaramillo et al., 2000). In Vitro Investigation on A.
muricata aqueous leaf extract it was performed against Haemonchus, a
gastrointestinal parasite. The result showed 89.08% and 84.91% toxicity against
larvae and eggs as assessed by larval motility egg hatch test. The
immobilization of adults worms within 8 to 9 hours of exposure to different
doses of the extract revealed a promising anthelmintic activity in the leaves
(Ferrcira et al., 2013).
2.5
Phytochemical
Extensive phytochemical evaluations on different parts of the A. Muricata plant
have shown the presence of various phytoconstituents and compounds including
alkaloids (AIKs) (Leboeaet al., 1990). Cydopeptides (PLs) and essential oils
(Kossouoh et al., 2007). However, Annona species, A. muricata have been
shown to be a generally rich source of annonaceous acetogenin compounds
(AGTs) (Rupprecht et al., 1990). The presence of different major minerals such
as K, Ca, Na, Cu, Fe and Mg suggest that regular consumption of the A.
muricata fruit can help provide essential nutrients and elements to the human
body (Gyamfet al., 2011).
2.6
Major Phytochemical Found in Plant
2.6.1 Alkaloid
These are the largest group of secondary chemical constituents made largely of
Anmmonia compounds comprising basically of Nitrogen base synthesized from
amino acid building blocks with various radicals replacing one or more of the
hydrogen atoms in the peptide ring and most containing oxygen. The
compounds have basic properties and are alkaline in reaction, turning red litmus
paper blue. Infact, one or more Nitrogen atoms that are present in a alkaloid,
typically as 10,20 and 30 amines, contribute to the basicity of the alkaloid. The
degree of basicity varies considerably, depending on the structure of the
molecule, and presence and location of the functional groups, (Sarker and
Nahar, 2007). They react with acids to form crystalline salts without the
production of water. Majority of alkaloids exist in solid such as a tropine, and
nitrogen (Firn, 2010).
Most alkaloids are readily soluble in alcohol and though they are aparingly
soluble in water, their salts are usually soluble. The solutions of alkaloids are
intensely better, these nitrogenous compounds function in the defense of plants
against herbivoures and pathogens, and are widely exploited as pharmaceuticals,
stimulants, narcotics, and poisons due to their potent biological activities. In
nature alkaloids exist in large proportions in the seed (Sarker and Nahar, 2007).
2.6.2 Flavonoids
Flavonoids are important group of polyphenols widely distributed among the
plant flora. Structurally they are made of more than one benzene ring in it’s
structure (a range of C15 aromatic compounds and numerous reports support
their use as antioxidants or free radical scavengers, the compounds are derived
from parent compounds known as (Flavans). Over four thousand flavonoids are
known to exist and some of them are pigments in higher plants. Quercetin,
Kaempterol and quercitrin are common flavonoids. Present in nearly 70% of
plants. Other group of flavonoids include flavones, dihydroflavons, flavans,
flavonols, anthoryanidins, anthoryanidins, proanthoryanidins, calchones. (Kar,
2007).
2.6.3 Saponins
The term saponin is drived from saponaria vaccaria (Quillaja saponarria)
Saponins therefore posses Soaplike behavior in water, I.e they produce foam, on
hydrolysis, an aglycone is produced which is called sapogenin. There are two
types of Sapogenin, steroidal and triterpenoidal. Usually, the sugar is attached at
uyC – 3 in Saponins, because in most Sapongernis there is a hydroxyl group at
C – 3. Saponins are extremely poisonous, as they cause heamolysis in blood and
are known to cause cattle poisoning (Kar, 2007). They possess a better and acrid
taste, besides causing irritation to mucus membrane. They are mostly
amorphous in nature, soluble in alcohol and water but insoluble in non-polar
organic solvents e.g benzene and n-hexane. Saponins are also important
therapeutically as they are shown to have hypolipidemic and anticancer activity
(Sarker and Nahare, 2007).
2.6.4 Tannins
These are widely distributed in plant flora. They are phenolic compounds of
high molecular weight. Tannins are soluble in water and alcohol and are found
in the root, bark, stem, seed and outer layers of plant tissue. Tannins have a
characteristics feature to tan, i.e to convert things into leather. They are acidic in
reaction and acidic reaction is attributed to the presence of phenolics or
carboxylic group (Kar, 2007). They form complexes with proteins,
carbohydrates, gelatin and alkaloids. Tannins are divided into hydrolysable
tannins and condensed tannins. Hydrolysable tannins, upon hydrolysis, produce
gall. Acid and ellagic and depending on the type of acid produced, the
Hydrolysable tannins are called gallotilic acid. Tannins are used as antiseptic
and this activity is due to the presence of the phenolic group. Common
examples of hydrolysable tannins include theaflavins (from tea): daidezein,
genistein and glycitein. Tannin – rich medicinal plants are used as healing
agents in a number of diseases. In Ayurveda, formulations based on tannin-rich
plants have been used for the treatment of diseases like levcorrhoea,
rhinnorhoea and diarrhea (Martinez et al., 2008).
2.6.5 Phenol
Phenolics, Phenol or polyphenolics (or Polyphenol extracts) are chemical
components that occur ubiquitously as natural colour pigments responsible for
the colour of fruit of plants. Phenolics in plants are mostly syntyhesized from
phenylalanine Via the action of phenylalanine ammonia lyase (PAL). They are
very important to plants and have multiple functions. The most important role
may be in plant defence against pathogens and herbivore predators, and thus and
applied in the control of human pathogenic infections. (Nascimento et al.,
2000). They are classified into (i) phenolic (ii) flon – flavonoid polyphenolics
(flavonones, flavones, xant hones and catechins) and (iii) Non – flavonoid
polyphenolies (affeic acid is regarded as the most common of phenolic
compounds distributed in the plant flora followed by chlorogenic acid known to
cause allergic dermatitis among humans (Kar, 2007). Phenolics essentially
represent a host of natural antioxidants used as nutraceuticals, and found in
apples, green tea, and red – wine for their enormous abiloity to combat cancer
and are also thought to prevent heart gilments to an appreciale degree and
sometimes are anti – inflammatory agents. Other example include flavones,
rutin, naringin, hesperidin and chlorogenic (Meijer man et al., 2006).
CHAPTER THREE
3.0
Materials and Method
3.1
Materials and Reagents Used
Water bath
Filter Paper
Volumetric Flask
Beakers
Hcl
2M HCl acid solution
Distilled water
Standard Tannin Solution
Sodium Carbonate Solution
Dragendorffs Reagent
1% Ammonium Hydroxide
10% acetic acid solution
Magnesium Metal.
3.2
Sample Collection
The fresh seeds of the sample (soursop were obtained from a local village of
Mbioto 1, Etinan Local Government Area and was taken to the biochemistry lab
in Akwa Ibom State Polytechnic for identification.
3.3
Sample Preparation
The sample (soursop seeds) were separated from the fruit (body), washed, and
sun dried for 5 days thereafter, grinded with blender into fine powder for further
analysis.
3.4
Phytochemical Screening
40g sample of dried soursop seed was milled into powdered form using an
electric blender. The sample was soak with 70% of ethanol with 30% of
distilled water to make up to 100% and was kept for 72 hours. Then after that, it
was filtered and the filtrate was put in container and put in water bath for
evaporation to take place.
The phytochemical screening of ethanolic extract of soursop seeds were
performed in order to ascertain the presence of its constitutions such as,
flavonoids, alkaloids, saponins, phenols, cardiac glycosides, and Tannins in the
ethanolic extract of soursop seeds, simple standard chemical testhave been
devised for screening.
3.5
Method of Phytochemical Screening
3.5.1 Test for Saponins
For the frothing tests, the method described by Wall et al., (1952 and 1954) was
used. About 0.5g of each plant extract was shaken with water in a test tube.
Frothing which persists on warming was taken as preliminary evidence for the
presence of saponins. In order to remove false – positive result, the blood
haemolysis test was performed on those extracts that frothed in water.
About 0.5g of each extract was boiled with 50ml phosphate buffer, pH 7.4 and
then allowed to cool and filtered. 5ml of the filtrate was passed fro 3hr, through
an asbestos disc (1.5mm thick and about 7mm in diameter), which had been
previously soaked with two or three drops of one (1) percent cholesterol in ether
and dried.
After filtration, the disc was wased with 0.5ml of distilled water, dried and
boiled in 20ml of oxylol for 2 hours to decompose the complex formed between
cholesterol on a 7 percent blood nutrient agar. Complete haemolysis of red
blood cells around the disc after 6 hours was taken as further evidence of
presence of Saponins (Trease and Evans).
3.5.2 Test for Flavonoids
Shinoda Reduction Test, 0.5g of the plant extract was Shaken with 5ml of conc.
HCl in a test tube and then, pieces of magnesium metals were added and
crimson colouration was observed.
3.5.3 Test for Alkaloids
0.5g of the extract was stirred with 5ml of 1 percent aqueous hydrochloric acid
on a steam bath. 1ml of the filtrate, 1ml portion was treated similarly with
Dragendorff’s reagent. And orange colouration with precipitate was observed.
3.5.5 Test for Cardiac Glycosides
a) Lieberman’s Test: 0.5g of the extract was dissolved in 3ml of choloroform
and filtered. Then 2ml of acetic anhydride was added to the filtrate then 2ml
of conc. H2SO4 was added carefully. Then pink colouration was observed at
the interphase.
b) Salkowski Test: 0.5g of the extract was dissolved in 3ml of chloroform and
was allow to cooled well and 2ml of conc. H2SO4 was then added carefully,
and Redish brown colouration was observed at the interphase.
c) Keller Kiliani Test: 0.5g of the extract was dissolved in 2ml of glacial
acetic containing one drop of ferric chloride solution. The 2ml of conc.
H2SO4 was added and browning at the interphase was observed.
CHAPTER FOUR
4.0
Result and Discussion
4.1
Result
Table 41: Phytochemical Screening of Ethanolic Extract of Soursop Seeds
Test
Observation
Inferences
Test for saponins frothing
Test
0.5g of the extract + 10ml of Frothing
persisted
for
distilled H2O and was shaken more than 10mins.
and
allowed
to
stand
++
for
10mins.
Test for Flavonoids Shinoda
Reaction Test
0.5g of the extract + 5ml of Crimson
conc.
HCl
+
pieces
colouration
of observed.
+++
magnesium metals.
Test
for
Tannins
Ferric
Chloride Test
0.5g of the extract + 10ml of Cloudy solution with ppt
distilled water and filtered. observed.
+
Filtrate + few drops of 5%
ferric chloride.
Test for Alkaloids
0.5g of the extracts + 5ml of Orang
colouration with
5% HCl and boiled and then ppt observed.
filtered. Filtrate + few drops of
dragendorff reagents.
Test for cardiac Glycosides
+++
Lieberman’s Test
(a)
Pink
0.5g of the extract of observed
colouration
at
the
3ml of chloroform and interphase
++
filtered. Filtrate + 2ml of
acetic anhydride + 2ml
of conc. H2SO4.
(b)
Salkowskis Test
Redish
0.5g of the extract + 3ml colouration
brown
at
the
at
the
++
of chloroform + 2ml of interphase
conc. H2SO4.
(c)
Kelter Kilians Test
Brownring
0.5g of the extract + 2ml interphase observed
of glacial acetic acid + 1
drop
of
5%
+
ferric
chlorides + 2ml of conc.
H2SO4.
Key:
4.2
+++ -
Abundantly Present
++
-
Moderately Present
+
-
Trace Present
Discussion
A. Muricata is a coveted troptical tree, and a wealth of phytochemical
investigation have been conducted for the fruit plant (seeds). In addition to
being an important source for the food industry and an indigenous medicinal
plant, A. muricata is proven to possess a wide spectrum of biological activities.
The result of the phytochemical screening carried on the seeds of soursop using
standard methods, (Ethanolic Seed extract) indicate the presences of some
phytochemicas.
Test for Saponins: Therefore posses soaplike behaviour in water i.e. they
produce foam, so when carried out test on saponins, frothing persisted was
found for more than 10mins and this indicated the presence of saponins.
Test for Flavonoids: Are important group of polyphenols among the plant flora
structurally, they are made of more than one benzene ring in its structure, so
when carrying test on flavonoids , when the pieces of magnesium metals was
added it evaporated and tend to be black but later extraction some crimson
colouration was observed.
Test for Tannins:They are phenolic compounds of high molecular weight,
tannins are soluble in water and alchohol and are found in the root, bart, stem
and outer layers of plant tissue. Tannins have a characteristic feature to tan, i.e.
to convert things into leather. They are acidic in reaction, when the extract was
added to 10ml of distilled water and filtered, the filtrate and drops of 5% of
ferric chloride, and cloud solution with precipitate was observed to prove that
tannins is present.
Test for Alkaloids: Alkaloids are largest group of secondary chemical
constituents made largely of ammonia compounds comprising basically of
nitrogen bases synthesis from amino acid building blocks. With various radical
replacing one or none of the hydrogen atoms in the peptide ring. The
compounds have basic properties are alkaline in reaction turning red litmus
paper blue. When the extract, 5ml of HCl and boiled and filtered, then filtrate
with few drops of dragendorff reagent, orange colouration with precipitate was
observed for evidence of Alkaloids present.
Test for Cardiac Glycosides: Glycosides is defined as the condensation
products of sugars (including polysaccharides) with a host of different varieties
of organic hydroxyl compounds (invariable monohydrate in character) in such a
manner that the hemiacetal entity of the carbohydrate must essentially take part
in the condensation. Glycosides are colorless, crystalline carbon, hydrogen and
oxygen – containing, chemically, glycosides contain a carbohydrate (glucose)
and a non-carbohydrate part (aglycone or genin).
Lieberman’s test when the extract and 3ml of chloroform and filtered, then, the
filtrate, 2ml of acetic anhydride and 2ml of conc. H2SO4 was added, pink
coloration was observed at the interphase and for
Salkowskis Test, Redish brown colouration at the interphase was observed.
Keller Kiliani Test, brownring colouration at the interphase was observed, all
the test was evidence for the present of cardiac Glycosides. The findings
indicate that soursop seeds are nutritional.
CHAPTER FIVE
5.0
SUMMARY, CONCLUSION AND RECOMMENDATION
5.1
Summary
The unripe fruit is green, hard both on the inside and outside and the pulp is
white. The fruit is used as natural medicine for arthritic pain, diarrhea, parasites,
skin rashes and worms.
And the crushed seeds are believed to have anthelmintic activities against
external and internal worms and parasites. A muricata is proven to posses a
wide spectrum of biological activities. Among all, the most promising activity
are found to be its anticancer and antiparasitic activity.
5.2
Conclusion
Medicinal plants are potent source of human health due to its compounds that is
responsible for its various. Pharmacological activities. Annona muricata, a
traditional medicinal plant (seeds) was investigated and showed that the
photochemical constituents and the bioactive compounds posses the medicinal
properties which makes them to be a potential species in the family of
Annonaceae.
5.3 Recommendation
Due to findings in this study, the following recommendations are made to create
awareness of phytochemical potency.
1. The seeds of Annona muricataare very rich, its has some chemical
compounds found on it, like saponins, flavonoids, Tannins, Alkaloids and
cardiac Glycosides which made the seed of the plant richly in medicinal
used.
2. Government should encourage the production of phytochemical supplement
(drug)
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