vol1-no1-5

JOURNAL OF CHEMICAL ENGINEERING VOL. 1, NO. 1, JANUARY 2012

38

Abstract— Leaves of Cassia occidentalis were extracted with

ethanol and water. The extracts were used to carryout

antimicrobial screening in vitro on staphylococcus aureus,

pseudomonas aeruginosa, Escherichia coli, salmonella typhi,

shigella spp. Chromatograhic separation was carried out on the

active extracts, and the efficacy of the resulting fractions was

tested against the susceptible organism. Some of the extracts

indicated significant inhibitory activity against the tested

organisms. General phytochemical screening was done on the

ethanol, water extracts and fractions. Ethanol extract revealed

the presence of Tannins, Saponins, Cardiac glycoside, Terpenoids

and Anthraquinones while the fraction revealed the presence of

Tannins, Terpenoid and Anthraquinones. This result might

explain the ethnobotanical use of the plant for the treatment of

dysentery, gastro internal disorder, constipation and Typhoid

fever.

.

Keywords: Phytochemistry ,Antimicrobial ,cassia occidentalis ,

plant ,Herbal

I. INTRODUCTION

lants serve as the basis of traditional medicine system for

thousands of years in Nigeria, India, China, Indonesia

[1]-[2]. In Yola, North eastern Nigeria. Butyrospermum

paradoxum, detatarium microcapum and Anogeneissus

Manuscript received January 27, 2012

I. S. Sadiq is with the Department of Natural Sciences, College of Science

and Technology, the Polytechnic of Sokoto State, PMB, 2356, Sokoto,

Nigeria. TEL: +2437035511295, email: [email protected].

M Shuabu is with the Department of Natural Sciences, College of Science


Nigeria.

A. B. Bello is with the Department of Natural Sciences, College of

Science and Technology, the Polytechnic of Sokoto State, PMB, 2356,

Sokoto, Nigeria.

S. G. Tureta is with the Department of Natural Sciences, College of


Sokoto, Nigeria.

T. Izuagie is with the Department of Natural Sciences, College of Science


Nigeria..

I. S. Sadiq is with the Department of Natural Sciences, College of Science


Nigeria.

S. Nasiru is with the Faculty of Veterinary Medicine, Usmanu Danfodiyo

University, Sokoto, Nigeria.

M. B. Kamaru is with the Department of Natural Sciences, College of


Sokoto, Nigeria.

Leiocarpus are said to have medicinal properties that are

effective in the management of diarrhea, dysentery, wound

infections, cough [3]-[4]. In Sokoto state, north western

Nigeria vernonia amygdalina leaves and securidaca

longipendunculata have also been reported to have medicinal

properties in the herbal remedies for leprosy, cough, gastro

internal disorder and toothache [5], [6]. The medicinal flora in

the tropical eco-region has a large abundance of plants that

provide raw materials for addressing medical disorders,

pharmaceutical and chemotherapeutic properties[7].

The active principle of many drugs found in plants is

phytochemical |6]. The medicinal value of these

phytochemicals is because of the presence of chemical

substance that produces definite physiological action on the

human body|8]. Some of the valuable one include:- Alkaloids,

tannins, saponins, glycosides, flavonoids, phosphorus and

calcium for cell growth, replacement, body building|8].

There are reports of antibiotics resistance of human

pathogens, to available antibiotics [9]-[10]. The Biomolecules

of these plants origin appears as alternative for the control of

these human pathogen [11] and their uses have been shown to

have scientific basis, chemicals compounds found in the

various species have different medicinal effects, ( e.g.

Alkaloids intercalate with DNA) |11]-[12].

The plant under investigation is Sanga – Sanga popularly

known by the Hausa speaking people, belongs to family

leguminosae, genus: cassia, species: occidentalis|13].

C occidentalis is a small tree that grow 5 – 8cm of south

America, including the Amazon. It is in the some genus as

Senna and sometimes called coffee senna.” Seeds pods long

are sometimes roasted and made a coffee-like beverage[14].

The cassia genus comprises some species of trees, shrubs,

vines and herbs with numerous species growing in the south

American rainforest, and tropics. Many species have been

medicinally, and these tropical plants have a history in natural

medicine as purgative and laxatives |15]. The main plant

chemicals in C. occidentalis include: achrosine, emodin,

anthraquinones, anthrones, apigenin, sitosterols, tannins and

xanthones. Toxicity studies on the aerial parts, leaves, roots of

C. occidentalis reported that various leaf and root extracts

given to mice (administered orally and injected at up to

500mg/kg) cause mortality [2]-[3]. Health practitioners today,

employed the uses in the remedy for bacteria/and fungal

infections. It is used as a broad spectrum internal and external

antimicrobial to treat bacterial and fungal infections. Also used

Phytochemistry and Antimicrobial Activities of

Cassia Occidentalis Used for Herbal Remedies

I. S. Sadiq, M. Shuaibu, A. B. Bello, S. G. Tureta A. Isah, T. Izuagie, S. Nasiru and M. B. Kamaru

P


39

for liver disorders (jaundice, lepatitis, cirrhosis, detoxification,

injury/failure, bile stimulant, etc). Also used in treatment for

intestinal worms, internal parasites, skin parasites. Moreso,

used as a cellular protector and preventative to cell damage

(immune, liver, kidney, cancer) |2].it is the intention of this

research work to identify the relevant phytochemical

compounds of the plant part that produces the antimicrobial

effect and to determine the antimicrobial activity of the plant

extract to correlate to its medicinal use.

II. MATERIALS AND METHODS

Sample Collection And Treatment

Fresh samples of plant leaves were sampled randomly from

different location in Sokoto states and was identified and

authenticated by the Botany Unit Department of Biological

Sciences of Usmanu Danfodiyo University Sokoto, a vouncher

specimen was deposited. The samples were air dried in the

laboratory and crushed using wooden pestle and mortal. The

crushed samples were sieved through 20-mesh sieve and the

fine powder was stored in a polythene bag for further use. The

powdered samples were used for the analyses.

Extraction:

100g of the powdered leaves was extracted with ethanol,

methanol and water using percolation process for 48hours.

Each extract was filtered and then evaporated to dryness using

rotatory evaporator. The resulting extract was used for the

following test. Each extract was subsequently weighed and the

percentage yield determined.

Column Chromatographic Separation

The lower end of a glass column 10cm long and 1.5cm in

internal diameter was plugged with glass wool. The material

was poured on the glass wool and air bubbles released was

trapped with the flat end of a packed rod. The column was

packed with wet silica gel by pouring the silica gel into the

column in a stepwise manner. The side of the column was

taped gently with a glass rod for compaction of the particles.

As silica gel settles, the column outlet was adjusted. Two (2g)

of each sample was drawn onto the adsorbent and eluted with

solvent mixture of ethanol and methanol in ratio (50:50)

v/v|16].

The fractions obtained was labeled as ; SL1, SL2, SL3,

SL4, SL5, and SL6 respectively.

Phytochemical Screening

The crude extract (ethanol) and the fractions obtained were

subjected to phytochemical analysis to determine the presence

of certain phytochemicals such as alkaloids, saponins, tannins,

flavonoids, steroids and cardiac glycosides using standard

methods [18], [19].

Antibacterial Species

Test organisms; this are cliinical isolates obtained from

patient attending the Usmanu Danfodiyo University Teaching

Hospital (UDUTH) Sokoto, Nigeria. These microorganisms

include staphylococcus aureus, Salmonella typhi, Escherichia

coli, shigella species and Pseudomonas aeruginosa. These

clinical isolate have been identified by standard procedure.

Preparation of Antibacterial Medium

28g of the nutrient agar was weighed and transferred into a

conical flask containing 100ml of distilled water. The mixture

was sterilized by steaming for 15 minutes. The medium was

allowed to cool, transferred into a sterilized petri dish up to the

mark, and allowed to solidify |18].

Antimicrobial Activity Test Using The Agar Well Diffussion

Method

Antibacterial activity of the ethanol, water extract and the

fractions was carried as reported |9], using agar well diffusion

method (ditch method). Four holes were made on sterilized

nutrient agar contain in a sterilized Petri dish. Each plate was

inoculated with bacterial species and the prepared

concentration of the extracts ( 30mg/ml 60mg/ml, 90mg/ml

and 120mg/ml) was transferred into the 6mm ditch holes

respectively. The plates were incubated at 37oC for 24 hours.

Clear zone of inhibition were measured after 24 hours of

incubation. The effect were compared with that of the standard

antibiotic Amoxilin(AMX),Septrin (SXT) and Ampiclox (A).

III. RESULTS AND DISCUSSION

Antibacterial activity

See table 1.2-1.3 for the presentation of zone of inhibition

(mm) for the leaves, and the fractions (SL1, SL2, SL3, SL4,

SL5 and SL6) respectively. General phytochemical analysis of

the six fractions revealed that SL1 and SL3 contained different

component.

The result show that these extracts are effective against all

of the test organisms. The highest activity (zone of inhibition

in diameter is about 18mm) was demonstrated by the ethanol

extract of Cassia occidentalis leaves against Salmonella typhi

while the lowest activity was 7mm by the water extract against

Shigella spp

On the other hand the ethanol and water extract were not

active against E.coli. at all concentration. The water extract

showed inhibition at lower concentration (30 and 60mg/ml)

against E. coli and Salmonella typhi.

These result. Suggested that antibacterial activity of Cassia

occidentalis leaves of ethanol and water extract against test-

organism were increase when used in higher concentration.

Like wise the fraction SL1 and SL3 were also active against

shigella and Salmonella typhi at higher concentration .

The ethanol and water extract s of the leaves of Cassia

occidentalis used in traditional Nigeria folk medicine were

analyzed for their antibacterial activity against five tested

organism (bacterial strains). The percentage yield of the

powdered sample of ethanol extract gave 12.5g and that of


40

water gave 7.5 g. The fractions when concentrated and

evaporated at 40 oC gave yield of 6.0mg, 5.2mg, 4.5mg,

4.2mg, 3.6ng and 3.0mg. The Rf values of compounds

separated were found to be 0.75, 0.82, 0.69 respectively. The

phytochemical constituent for the ethanol and water extract

revealed the presence of tannins, cardiac glycosides, Saponins,

and Anthraquinone respectively, while the fractions revealed

the presence of tannins, terpenoids and anthraquinones

respectively. These phytoactive compound are well known for

their wide pharmacological activities ranging from

antibacterial and antifungal |21]. The result of this study show

that the ethanol and water extracts inhibited the growth of

various species of gram-negative bacteria, the ethanol and

water extract show slightly better killings action which means

that either ethanol or water extract could be used often,

however staphylococcus aures was resistant to both ethanol

and water extract. That probably could be due to cell

membrane permeability or due to genetic factors.

On the other hand the hydrophobic character of phenolic

compound can potentially impair cellular function and

membrane integrity |17]. The capacity of phenolic compounds

to chelate transition metals also lower the reactivity of metals

ion by forming an inert metals-ligand complex. Chelation of

transition metals, such as iron and copper, reduces

bioavailability for bacterial growth|19]. The diameter of

inhibition zone around the most active extract were

comparable with the standard antibiotics used as a positive

control. The completely gram-negative bacterial were resistant

to Amoxilin except salmonella typhi.

These result were found to be in accordance with the use of

a decoction of fresh leaves of Cassia occidentalis to promote

its usage in the treatment for typhoid fever, gastrointestinal

disorder, and as body wash in folk medicine .

IV. CONCLUSION

The result of this study indicated that the parts of plant

contain some major bioactive compound that inhibits the

growth of microorganism thereby proving very effective

source of derived drugs. Since this plant proved very active.

This agrees with the report that the leave are used for the

remedy of dysentery, constipation and gastro internal disorder

and treatment of typhoid fever. Therefore, it is recommended

that the different component detected in the plant should be

isolated and then be mixed in different proportion and the

mixture tested against the susceptible microorganisms in order

to arrived at the most potent structure.

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[2] Bin-Hafeez, B. and Hussaini A.S (2001) Protective effect of Cassia

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[10] Kubmarawa D. Ajoku, Enwerem, M. and Okorie, D. (2007).

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M. Hassan and R. Atiko (2008). The phytochemistry and

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Antimicrobial Agents. Microbes and infections. 4:613-620.

[13] Mitsuyama, J., Hiruma,, R, Yamaguchi, A. and T Sawai (1987).

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Edition Bradford Publishing House, Germany. PP 1 – 10.

[17] Raccach, M,(1984). The antimicrobial activity of phenolic antioxidant,

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41

Table 1.0 Ethanol, and water extracts of leaves of cassia

occidentalis

Phytochemical

compounds

Ethanol Methanol Water

Alkaloids - - -

Tannins + + +

Flavonoids - - -

C. Glycosides + + +

Saponins + + +

Anthraquinone + + +

+= Present, - = Absent

Table 1.1 Fractions for leaves of cassia occidentalis

Phytochemicals

Fractions

SL1 SL2 SL3 SL4

SL5 SL6

Alkaloids - - - - - -

Flavonoids - - - - - -

C. Glycosides - - - - - -

Terpenoids - - + - - -

Saponins - - - - - -

Tannins + - - - - -

Anthraquinone + - + - - -

Key: C = cardiac

Antibacterial activity

See table 1.2-1.3 for the presentation of zone of inhibition (mm) for the leaves, and the fractions (SL1, SL2, SL3, SL4, SL5 and SL6)

respectively. General phytochemical analysis of the six fractions revealed that SL1 and SL3 contained different component.

Table 1.2 Antibacterial Activities of Ethanol and water Extracts of Cassia .Ocidentalis zone of inhibition (mm) at various concentration

(mg/ml).

ETHANOL WATER ANTIBIOTIC DISC

Bacterial Spp. 30 60 90 120 30 60 90 120 AMX SXI A

Pseudomonas .A

E. Coli

Shigella Spp

Staph aureus

Salmonella Typhi

7 7 9 R

R 9 10 12

R 11 13 14

R R R R

9 12 13 18

R 8 10 12 R 24 10

8 9 9 10 R R 18

R R 7 7 R 15 R

R R R R R R 10

8 9 15 17 20 R R

Table 1.3 Inhibition Zone (mm) of Cassia occidentalis fraction (SL1 and SL3) at various concentration (mg/ml).

ETHANOL WATER ANTIBIOTIC DISC

Bacterial Spp. 30 60 90 120 30 60 90 120 AMX SXI A

Pseudomonas .A

E. Coli

Shigella Spp

Staph aureus

Salmonella Typhi

R R R R

R R 7 12

R R 9 14

R R R R

R 7 9 13

R R R R R 15 R

R R R R R R R

R 8 8 11 R 15 R

R 7 7 R R R R

R 9 11 13 18 R R

A =aeruginosa E= Escherichia Spp. =Specie

R- Resistance Any Zone of inhibition greater than 6mm shows activity or sensitivity.

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