PHYTOCHEMICAL PROFILING AND ANTIOXIDANT

ANALYSIS OF SYZGIUM JAMBOS L. (ALSTON)

BY

OYEBAYO OLALEKAN AYOOLA

MATRIC NO: SCI/17/18/0995

A Research Project Submitted to the Department of Plant Science, Faculty of

Science, Olabisi Onabanjo University, Ago-Iwoye, Ogun State.

In Partial fulfillment of the Requirements for the Award of Bachelor of Science

Degree (B.Sc.) IN PLANT SCIENCE

NOVEMBER, 2021
 TABLE OF CONTENTS
Pages
Title
Abstract i
Certification ii
Dedication iii
Acknowledgements iv
Table of Contents v
CHAPTER ONE: INTRODUCTION
1.1 Background to the Study
1.2 Statement of Problems
1.3 Aim and Objectives
1.4 Justification of the Study
CHAPTER TWO: LITERATURE REVIEW
2.1 Secondary metabolites in plants
2.2. Extraction of Phenolic Compounds Using Solvents
2.3 Antioxidants present in plants
2.4 Antioxidants present in Syzygium jambos
2.5 Empirical Review
CHAPTER THREE: MATERIALS AND METHODS
CHAPTER FOUR: RESULTS
CHAPTER FIVE: DISCUSSION
CHAPTER SIX: CONCLUSION AND RECOMMENDATION
List of Tables
List of Figures (Bar chart, Pie chart etc.)
List of Plates
REFERENCES

APPENDICES
 CHAPTER ONE

1.0 INTRODUCTION

1.1 Background to the Study

Fruits and vegetables have been proven to have several antioxidant compounds which, this

compounds includes phenolics, carotenoids, anthocyanins, and tocopherols (Koffi, et al.,

2010). Approximately 20% of known plants have been used in pharmaceutical studies, which

have been impacting the healthcare system greatly in positive ways. This plants have been

known to be a great remedies in solving health issues, such as treating cancer and harmful

diseases (Anokwuru, et al., 2011).

Plants are able to manufacture depots of a great amount of diverse bioactive compounds.

High concentrations of phytochemicals, which may fight and protect against free radical

damage, accumulate in fruits and vegetables (Ballard, et al., 2010). Plants which are found to

contain phytochemicals are beneficial to the human body because they act as natural

antioxidants in the human body system. (Li, H, et al., 2012). Various studies have shown

that many plants are rich source of antioxidants. For instance, vitamins A, C, E, and phenolic

compounds such as flavonoids, tannins, and lignins, found in plants, all act as antioxidants

(Koffi, et al 2010). There are several health benefits found in the consumption of vegetables

and fruits, a result of medicinal properties and high nutritional value (Anokwuru, et al.,

2011). Antioxidants do control and reduce the oxidative damage in foods by delaying or

inhibiting oxidation caused by reactive oxygen species (ROS), ultimately increasing the

shelf-life and quality of these foods (Ballard, et al., 2010). Beta carotene, ascorbic acid, and

many phenolics play dynamic roles in delaying aging, reducing inflammation, and preventing

certain cancer. Increasing the consumption of fruits and vegetables has been recommended

by many agencies and health care systems throughout the world (Anokwuru, et al., 2011).
The rose apple, Syzygium jambos (syn. Eugenia jambos L.), is a large shrub or small to

medium in size tree, known to grow originally in Southeast Asia, but now widely distributed

in the tropics. The plant has been traditionally used for its antipyretic and anti-inflammatory

properties and to treat hemorrhages, syphilis, leprosy, wounds, ulcers, and lung diseases. In

Indo-China, all plant parts are employed to treat digestive tract and tooth disorders. The

leaves decoction is used as antirheumatic, diuretic, and to relieve sore eye conditions (Kuiate

et al., 2007; Nawwar et al., 2016).

Syzygium jambos belongs to The Myrtaceae, is in the major groupa family of flowering

plants which comprisesd about 100 genera and 3000 species throughout the tropics and

subtropics especially in Australia . The generic name is derived from the Greek syzygios

(paired), on account of the leaves and twigs that in several species grow at the same point . In

Malaysia, Myrtaceae consists of about 9 genera and 210 species which are distributed from

the lowland to mountain forests (Kamal, 2014). The leaves are simple, estipulate,

oppositeopposite, or alternate, and finely dotted with oil glands. The flowers are small to

medium sized, regular, and bisexual. All the Malaysia’s species are evergreen but they flush

new leaves and flowers seasonally. There are eight other genera in Myrtaceae family of

Malaysia such as Syzygium (Eugenia), Baeckea (Chuchor atap), Rhodomyrtus (Kemunting),

Leptospermum (China maki), Melaleuca (Gelam), Tristania (Pahlawan/Pelawan),

Rhodamnia (Mempoyan), Pseudoeugenia, and Decaspermum. This family has commercial

importance for its flower, edible fruits, oil-yielding glands, and timbers. (Kamal, 2014)

Syzygium jambos may be merely a shrub but is generally a tree reaching 7.5-12 m in height,

and has a dense crown of slender, wide-spreading branches, often the overall width exceeding

the height. The evergreen leaves are opposite, lanceolate or narrow-elliptic, tapering to a
point, 10-22 cm long, and 2.5-6.25 cm wide; somewhat leathery, glossy, darkgreen when

mature, rosy when young. (Orwa et al. 2009).

The flowers are creamy-white or greenish-white, 5-10 cm wide, consisting mostly of about

300 conspicuous stamens to 4 cm long, a 4-lobed calyx, and 4 greenish-white, concave petals.

There are usually 4 or 5 flowers together in terminal clusters.

Capped with the prominent, green, tough calyx, the fruit is nearly round, oval, or slightly

pear-shaped, 4-5 cm long, with smooth, thin, pale-yellow or whitish skin, sometimes pink-

blushed, covering a crisp, mealy, dry to juicy layer of yellowish flesh, sweet and resembling

the scent of a rose in flavour, Plate 1 and 2 (Orwa et al.2009).

In the hollow center of the fruit, are 1-4 brown, rough-coated, medium hard, more or less

rounded seeds, 1-1.6 cm thick, which loosen from the inner wall and rattle when the fruit is

shaken. Fragments of the seed coat may be found in the cavity (Plates 1 and 2). See Plate 1

and 2.

The generic name is derived from the Greek syzygios (paired), on account of the leaves

and twigs that in several species grow at the same point.

The phytochemistry and pharmacology of several members of this genus (including S.

cumini, S. aqueum, S. samarangense, S. aromaticum, and S. jambolanum) have been

extensively investigated. In traditional medicine, some of them have been used to treat

several disorders, such as hemorrhage, dysentery and gastrointestinal disorders, as well as

diabetes and inflammation as these extracts possess antifungal, antimicrobial,

antihypertensive, analgesic and antiviral (against herpes virus) properties (Raga et al., 2011;

Sharma, Kishore, Hussein, & Lall, 2013).

1.2 Statement of Problems

Infectious diseases cause 15 million deaths every year, accounting for about 27.12% of

deaths worldwide (Dye, 2015). Multidrug- resistant (MDR) bacteria are responsible for

therapeutic failures, leading to an increase disease burden. Despite the various

technological and medical-pharmaceutical advances, MDR bacteria still remain a major

cause ofuse of plants morbidity and mortality globally. The search for new antibacterial

substances should therefore take into account the development of resistance by

pathogenic bacteria. With regard to the high diversity of secondary metabolites in plant

kingdom, botanicals constitute a good reservoir for drug discovery to combat MDR

bacteria (Dzotam, et al 2016). Also, the loss of efficacy of several antibiotics and the

scarcity of new antibacterial agents propel the search for substances capable of

restoring the activity of antibiotics.

African medicinal plants have previously shown efficiency against MDR bacteria with

some of them being able to modulate the activity of antibiotics. Some of these plants

include Xanthosoma mafaffa, Moringa oleifera, Passiflora edulis, Anthocleista

schweinfurthii, Nauclea latifolia, Zehneria scabra, Nauclea pobeguinii, Catharanthus

roseus, Croton macrostachys, Paullinia pinnata, Albizia adianthifolia, Alchornea

laxiflora, Laportea ovalifolia , Mangifera indica , Ricinodendron heudelotii , and

Euphorbia prostrata (Dzotam, et al 2016; Djeussi, et al 2015).

In our continuous search for new botanicals to combat MDR bacteria as well as

potentiate the activity of antibiotics, we targeted another African plant, Syzygium

jambos (L.) Alston. (Myrtaceae). This plant is used traditionally to treat abdominal

pain, diarrhea, amenorrhea, pernicious attacks, epilepsy, asthma, bronchitis, diuretics,

rheumatism, small-pox and eye irritation (Seukep, et al., 2016), respiratory disorders,

eczema, malaria, and infectious diseases. Previous studies have reported the
antibacterial effects of extracts of bark, leaves, and seeds of Syzygium jambos against

sensitive phenotypes (Seukep, et al., 2016; Tchinda, 2017).

1.3 Aim and Objectives

The main aim of the study is to estimate the total phenol, the total flavonoids and antioxidant

activities of ethanol extract of Syzygium jambos.

Specific objectives: with the following objectives:

(i) To evaluate antioxidant activity of Syzygium jambos

(ii) To quantify total phenol in ethanol extract of Syzygium jambos

(iii) To ascertain the total flavonoids present in ethanol extract of Syzygium jambos

1.4 Justification of the Study

In recent years, due to toxicological concerns associated with the use of synthetic substances

in food and increasing awareness about natural foods, there has been an increased interest in

the use of natural substances as food preservatives and antioxidants (Fankam, et al., 2017).

Flavonoids, tannins, anthocyanins and other phenolic constituents present in food of plant

origin are potential antioxidants (Voukeng, et al., 2017).

Food rich in antioxidants plays an essential role in the prevention of some cancers and

cardiovascular and Alzheimer’s diseases. Therefore, the search for antioxidants,

hypoglycemic and anticancer agents in vegetables, fruits, tea, spices and medicinal herbs

have attracted increasing attention (Murugan, et al., 2011). There have been a general believe

that the human resources and ethno-medical knowledge endemic plants will be the basis for

the development of raw materials for natural herbal medicines by utilizing local resources.

Until recently, there has been a lack of information about the potential bioactivity of

Syzygium jambos and the content of chemical compounds contained therein (Fankam, et al.,
2017; Murugan, et al., 2011). This study aimed to inquire aboutinvestigate the antioxidant

activities and to perform screen phytochemicals screening ofin ethanolic extracts of

Syzygium S. jambos using ethanol extract.

CHAPTER TWO

LITERATURE REVIEW

2.1 Secondary metabolites in plants

Phytonutrients or phytochemistry originating from the plants results in various activities that

benefit the health of humans and the plants themselves. These compounds are known as

secondary metabolites such as alkaloids, terpenoids, steroids, and flavonoids. Among them,

flavonoids, sometimes called phenolic compounds, exhibit antioxidant properties, which

allow them to produce anti-inflammation, antiallergenic, antivirus, antiaging, and

anticarcinogenic effects (Agati, et al., 2012; Panche, et al., 2016).

Two groups of antioxidants (i.e.,that is natural antioxidants and synthetic antioxidants) are

often employed in medicine and food especially those containing fats and oils to prevent

oxidation. Prominent synthetic antioxidants are butylated hydroxyanisole (BHA) and

butylated hydroxytoluene (BHT), which have been extensively used in the food, cosmetic,

and therapeutic industry. However, these synthetic antioxidants are carcinogenic, highly
volatile, and uinstable at high temperature, potentially causing various fatal diseases.

Therefore, a natural antioxidant is required as an alternative that is appropriate for dietary

intake, shows no hazardous effects to the human body, and is obtainable at a low cost (Lobo,

et al., 2010).

2.2. Extraction of phenolic compounds using solvents

Scientists have studied and analysed the impact of different types of solvents, such as

methanol, hexane, and ethyl alcohol, for the purpose of antioxidant extraction from various

plants parts, such as leaves and seeds. In order to extract different phenolic compounds from

plants with a high degree of accuracy, various solvents of differing polarities must be used

(Koffi, et al., 2012). Moreover, scientists have discovered that highly polar solvents, such as

methanol, have a high effectiveness as antioxidants.

Anokwuru et al. (2011) reported that acetone and N,N dimethylformamide (DMF) are highly

effective at extracting antioxidants, while Koffi et al. found that methanol was more effective

in at a large amount of phenolic contents from walnut fruits when compared to ethanol

(Koffi, et al., 2012). It has been reported that ethanolic extracts of Ivorian plants extracted

higher concentrations/amount of phenolics compared to acetone, water, and methanol

(Ballard, et al., 2012).

Multiple solvents have been commonly used to extract phytochemicals, and scientists usually

employed a dried powder of plants to extract bioactive compounds and eliminate the

interference of water at the same time. Solvents used for the extraction of biomolecules from

plants are chosen based on the polarity of the solute of interest. A solvent of similar polarity

to the solute will properly dissolve the solute (Koffi, et al., 2012).

Multiple solvents can be used sequentially in order to limit the amountquantity of analogous

compounds in the desired yield. The polarity, from least polar to most polar, of a few
common solvents is as follows: Hexane < Chloroform < Ethylacetate < Acetone < Methanol

< Water. (Ammar, et al., 2017)

2.3 Antioxidants present in plants

Many antioxidant compounds can be found in fruits and vegetables including phenolics,

carotenoids, anthocyanins, and tocopherols. Approximately 20% of known plants have been

used in pharmaceutical studies, impacting the healthcare system in positive ways such as

treating cancer and harmful diseases (Naczk, et al., 2006).

Plants are able to produce a large number of diverse bioactive compounds. High

concentrations of phytochemicals, which may protect against free radical damage,

accumulate in fruits and vegetables. Plants containing beneficial phytochemicals may

supplement the needs of the human body by acting as natural antioxidants (Ammar, et al.,

2017). Various studies have shown that many plants are rich source of antioxidants. For

instance, vitamins A, C, E, and phenolic compounds such as flavonoids, tannins, and lignins,

found in plants, all act as antioxidants. The consumption of fruits and vegetables has been

linked with several health benefits, a result of medicinal properties and high nutritional value

(Koffi, et al., 2012).

Antioxidants control and reduce the oxidative damage in foods by delaying or inhibiting

oxidation caused by reactive oxygen species (ROS), ultimately increasing the shelf-life and

quality of these foods. Beta carotene, ascorbic acid, and many phenolics play dynamic roles

in delaying aging, reducing inflammation, and preventing certain cancers (Boots, et al.,

2006). Increasing the consumption of fruits and vegetables has been recommended by many

agencies and health care systems throughout the world (Ammar, et al., 2017).

2.4 Antioxidants present in Syzygium jambos

The chemical composition and antioxidant activity of S. jambos fruits have been also studied

recently and tannins extracted from S. jambos fruit showed a very good DPPH radical

scavenging activity and ferric reducing/antioxidant property (Bajpai, et al., 2005). According

to nutritionists, the fruit of S. jambos is rich in carbohydrates, minerals and vitamins, with

glucose and fructose as principal sugars. The fruit is also rich in minerals such as manganese,

zinc, iron, calcium, sodium and potassium.

The ripe S. jambos fruit has many therapeutic properties such as liver stimulation, digestive,

carminative, coolant and hypoglycemic effects. Its leaves contain essential oils with a

pleasant odour. The oil contains terpenes, dipentenes, sesquiterpenes, ellagic acid,

isoquercitin, quercetin, kaempferol and myricetin in different concentrations (Abhishek, et

al., 2011). The barks, leaves and seeds extracts of S. jambos have been reported to possess

anti-inflammatory, and antidiarraheal effects. The leaves have been extensively used to treat

diabetes and constipation, fever, gastropathy, strangury and dermopathy (Benherlai, et al.,

2007).

2.5 Empirical Review

Abdullah and Jamil, (2020) studied pPhytochemical screening contents and antioxidant

activities of mMethanol extracts from eight Syzygium species, The dried leaves, twigs, and

heartwoods of Syzygium species were ground into powdered form and extracted using cold

extraction method with MeOH solvent. The extracts were filteredfiltered, and the solvent was

removed under vacuum using a rotary evaporator to obtain the MeOH extracts. The

preliminary phytochemical screening of the MeOH crude extracts were conducted in order to

determine the presence of secondary metabolites in all parts of eight Syzygium plants (S.

filiforme var. filiforme, S. papillosum, S. griffithii, S. pseudoformosum, S. claviflorum var.

claviflorum, S. glaucum, S. syzygioides, and Eugenia Sp. 57). Analysis of different parts of

the plant extracts (leaves, twigs, and heartwoods) revealed the presence of alkaloids,

flavonoids, phenols, steroids, terpenes, and tannins except saponins.

Previous studies reported that several Syzygium plant species have antibacterial activity

properties (Mawan et al. 2018). S. polyanthum infusion extract of antibacterial exhibited

activity against Bacillus cereus, Salmonella thypi, Trichophyton mentagrophytes, and

Candida albicans (Kusuma et al. 2011) and the methanol extracts from S. polyanthum fruit

have antibacterial activity against E. coli(Mawan et al. 2018).

Fitri et al. (2017) reported that the ethanol extract of S. polyanthum leaves has antibacterial

activity against Shigella dysenteriae. Several other Syzygium species were reported to have

antibacterial action such as the ethanol, ethyl acetate and water extracts from S. cumini can

inhibit the growth of E. coli, Salmonella typhi, Pseudomonas aeruginosa, and S. aureus (Kaur

et al. 2017; Mohamed, Ali and El-Baz 2013; Shafi et al. 2002).

Methanol and water extracts in S. jambolanum has been reported to inhibit the growth of

several Salmonella typhimurium, Pseudomonas aeruginosa, Klebsiella pneumoniae, E. coli,

Bacillus subtilis, S. aureus (Chandrasekaran and Venkatesalu 2004; Mohanty and Cock

2010). Moreover, it has been shown that the leaf extract of Syzygium jambos L. is potent

against the pathogenic Propionibacterium acnes (Sharma et al. 2013b). Djoukeng et al. (2005)

reported that the leave extracts of S. guineense have significant antibacterial activity against

E. coli, Bacillus subtilis dan Shigella sonnei.

Gavillán-Suárez et al. (2015) studied chemical profile and in vivo hypoglycemic effects of

syzygium jambos, costus speciosus and tapeinochilos ananassae plant extracts used as

diabetes adjuvants in puerto rico. In his methodology, Phytochemicals in methanolic and

aqueous extracts were analyzed by thin layer chromatography (TLC). Alkaloids

(Bromocresol green, λ = 470 nm), flavonoids (AlCl3, λ = 415 nm), saponins (DNS, λ = 760

nm), tannins (FeCl3/K4Fe(CN)6, λ = 395 nm) and phenolics (Folin-Ciocalteau, λ = 765 nm)

were quantified. Male C57BLKS/J (db/db) and C57BL/J (ob/ob) genetically obese mice were

orally gavaged with aqueous extracts of lyophilized plant decoctions for 10weeks. Their

results shows that T. ananassae had significantly greater amounts of flavonoids and tannins,

while S. jambos showed the greatest concentration of phenolics and C. speciosus exhibited

higher amounts of alkaloids. C57BLKS/J db/db treated with plant extracts show better

glucose modulation when the extracts are administered in complement with an insulin

injection. Finally, C57BL/J ob/ob mice on T. ananassae and S. jambos treatments show better

blood glucose modulation over time.

List of plate
Plate 1: Syzygium jambos (Agroforestry Database 4.0)
Plate 2: Syzygium jambos (Agroforestry Database 4.0)
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