JOURNAL OF CHEMICAL ENGINEERING
1, JANUARY 2012 38
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
 Leiocarpus are said to have medicinal properties that are
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
P 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: shinasharon@yahoo.co.uk.
M Shuabu is with the Department of Natural Sciences, College of Science
and Technology, the Polytechnic of Sokoto State, PMB, 2356, Sokoto,
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
Science and Technology, the Polytechnic of Sokoto State, PMB, 2356,
Sokoto, Nigeria.
T. Izuagie is with the Department of Natural Sciences, College of Science
and Technology, the Polytechnic of Sokoto State, PMB, 2356, Sokoto,
Nigeria..
I. S. Sadiq is with the Department of Natural Sciences, College of Science
and Technology, the Polytechnic of Sokoto State, PMB, 2356, Sokoto,
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
Science and Technology, the Polytechnic of Sokoto State, PMB, 2356,
Sokoto, Nigeria.
VOL. 1, NO.
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
JOURNAL OF CHEMICAL ENGINEERING
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
VOL. 1, NO. 1, JANUARY 2012 39
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
JOURNAL OF CHEMICAL ENGINEERING
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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JOURNAL OF CHEMICAL ENGINEERING VOL. 1, NO. 1, JANUARY 2012 41

Table 1.0 Ethanol, and water extracts of leaves of cassia Phytochemicals Fractions
occidentalis SL1 SL2 SL3 SL4
Phytochemical Ethanol Methanol Water SL5 SL6
compounds Alkaloids - - - - - -
Alkaloids - - -
Flavonoids - - - - - -
Tannins + + +
C. Glycosides - - - - - -
Flavonoids - - - Terpenoids - - + - - -
C. Glycosides + + + Saponins - - - - - -

Saponins + + + Tannins + - - - - -
Anthraquinone + - + - - -
Anthraquinone + + +
Key: C = cardiac
+= Present, - = Absent

Table 1.1 Fractions for leaves of cassia occidentalis

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 7 7 9 R R 8 10 12 R 24 10
E. Coli R 9 10 12 8 9 9 10 R R 18
Shigella Spp R 11 13 14 R R 7 7 R 15 R
Staph aureus R R R R R R R R R R 10
Salmonella Typhi 9 12 13 18 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 R R R R R R R R R 15 R
E. Coli R R 7 12 R R R R R R R
Shigella Spp R R 9 14 R 8 8 11 R 15 R
Staph aureus R R R R R 7 7 R R R R
Salmonella Typhi R 7 9 13 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.