Phytochemical and Anti-Microbial Studies on the Leaves Extracts of Cardiospermum halicacabum Linn

Abstract

The phytochemical screening and In-vitro anti-microbial activity leaves of cardiospermum halicacabum linn (Sapindaceae) were investigated. The preliminary phytochemical analysis revealed the presence of alkaloids, carbohydrates, proteins and saponins. The extracts exhibited marked anti-microbial activity against both Gram +ve and Gram-ve bacteria. When the concentration of the extracts was increased the zone of inhibition also increased.

Full text

Advances in Biological Research 6 (1): 14-18, 2012
ISSN 1992-0067
© IDOSI Publications, 2012
DOI: 10.5829/idosi.abr.2012.6.1.56393
Corresponding Author: T. Deepan, Research Labortary, GIET School of Pharmacy,
Rajahmundry-533294, Andhra Pradesh, India.
14
Phytochemical and Anti-Microbial Studies on the Leaves
Extracts of Cardiospermum halicacabum Linn.
T. Deepan, V. Alekhya, P. Saravanakumar and M.D. Dhanaraju
11 2 1
GIET school of pharmacy, Rajahmundry, Andhrapradesh, India
1
Anna university, Trichy, Tamilnadu, India
2
Abstract: The phytochemical screening and In-vitro anti-microbial activity leaves of cardiospermum
halicacabum linn (Sapindaceae) were investigated. The preliminary phytochemical analysis revealed the
presence of alkaloids, carbohydrates, proteins and saponins. The extracts exhibited marked anti-microbial
activity against both Gram +ve and Gram-ve bacteria. When the concentration of the extracts was increased the
zone of inhibition also increased.
Key words: Cardiospermum halicacabum Linn Phytochemical Studies Anti-Microbial Activity
INTRODUCTION Cardiospermum halicacabum (Linn), family
Plant-based drugs have been used worldwide in
traditional medicine for the treatment of various diseases.
Approximately 60% of world’s population still relies on
medicinal plants for their primary healthcare. According to
a survey by NCI, USA, 61% of the 877 small-molecule new
chemical entities introduced as drugs worldwide during
1981-2002 were inspired by natural products [1] Plant
species still serves as a rich source of many novel
biologically active compounds, as very few plant species
have been thoroughly investigated for their medicinal
properties [2]. Thus, there is renewing interest in
phytomedicine during last decade and nowadays many
medicinal plant species are being screened for
pharmacological activities [3].
When we reviewed the number of articles published
on the antimicrobial activity of medicinal plants in
PubMed during the period between 1966 and 1994, we
found 115; however, in the following decade between
1995 and 2004, this number more than doubled to 307.
Many focus on determining the antimicrobial activity of
plant extracts found in folk medicine [4], essential oils [5]
or isolated compounds such as alkaloids [6], flavonoids
[7], sesquiterpene lactones [8], diterpenes [9], triterpenes
[10] or naphtoquinones [11] among others. Some of these
compounds were isolated or obtained by bio-guided
isolation after previously detecting antimicrobial activity
on the part of the plant [12].
Sapindaceae, is a deciduous, branching, herbaceous
climber, which is distributed through out the plains of
India. The whole plant has been used for several centuries
in the treatment of rheumatism, stiffness of limbs, snake
bite; its roots for nervous diseases, as a diaphoretic,
diuretic, emetic, emmenagogue, laxative, refrigerant,
stomachic and sudorific; its leaves and stalks are used in
the treatment of diarrhea, dysentery and headache and as
a poultice for swellings. Phytochemical constituents such
as flavones, aglycones, triterpenoids, glycosides and a
range of fatty acids and volatile ester have been reported
from the various extracts of this plant [13]. Most likely
confused with: Physalis spp. (ground cherry), Clematis
occidentalis, Clematis virginiana, Campsis radicans,
Adlumia fungosa [14]. The aim of the study was to show
that leaves of cardiospermum halicacabumlinn
linn…have antimicrobial activity and preliminary
phytochemical analysis was also evaluated
MATERIALS AND METHODS
Materials: Dried leaves powder of C. halicacabum linn,
ethanol (SD Fine Chemicals, Mumbai, India), chloroform
water (SD fine, Mumbai, India. ), beef extract (Merck Ldt,
India), acid hydrolysate casein (biomark laboratories,
Maharashtra, India), starch (rajlal and ons ndore, India),
agar-agar (Marine Chemicals, Kerala, India), sodium
hydroxide (s.d. Fine Chem.), hydrochloric acid (Merck Ltd,
India), ciprofloxacin, distilled water.

Advan. Biol. Res., 6 (1): 14-18, 2012
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Plant Collection and Identification: Leaves of the plant C. plate. oth aqueous and alcoholic extracts (sample) in
halicacabum linn collected 20 km away from erambalur, different concentrations were placed. Then the Petridish
Tamilnadu, India. This collection of the plant was incubated at 37°C for 24 hrs [20, 21]. Ciprofloxacin
authenticated by Dr. Kosiba B.S., M.S, (Govt.Regd. no: (50mg/ml) was used as a standard drug to compare
1209) Assistant siddha medical officer, Government different concentrations (50mg/ml,75mg/ml and 100mg/ml)
district head quarters, perambalur-621 212. of the sample.
Extraction of Plant Material: The leaves of RESULT AND DISCUSSION
cardiospermum halicacabum Linn were dried in the
shade. Then the dried leaves were powdered to get a The antimicrobial activity of the leaf extract was
course powder. About 25g of dry powder was extracted assayed by Cup plate or cylinder plate method method
with 200ml of ethanol (95%) or 200ml distilled water and against 3 bacterial species S. aureus, B. substilis and E.
10ml chloroform by hot continuous percolation method coli [19]. Table 2 shows the microbial growth inhibition of
using soxhlet apparatus. The extractions were continued both aqueous and methanol extracts of the screened
for 72 hrs at 50°C. The extract were filtered and plantspecies. The aqueous extract showed antimicrobial
concentrated to a dry mass by using vacuum distillation activity (Cardiospermum helicacabum linn…).
method. The extract stored in desiccators [15]. The methanol extracts of the investigated plants
Qualitative Phytochemical Evaluation: The shade dried negative K.pneumoniae. Similar results were also reported
powder and various extracts of the leaves of C. byParekh, J., et al. (17), Babu Ananth, D. The studied
halicacabum linn were subjected to chemical tests for its plant were most active against E. coli. The extracts of
active constituents. Identification of the chemical Preliminary phytochemical analysis revealed the presence
constituents was carried out on the same extracts used in of alkaloids (+ve test for alkaloids and saponins. The
pharmacological tests according to the methodology other secondary metabolites like saponin, protein, lignin,
proposed by Makonnen et al. [16] and parekh and Sumitra etc. were present in trace amounts in some of the plant
[17]. (Table 1). It is not surprising that there are differences in
Cup Plate or Cylinder Plate Method: Cup plate method thephytochemical properties and varying among
using Mueller-Hinton agar medium was employed to concentration.. The drying process may havecaused
study the antibacterial activity of the extracts against S. conformational changes to occur in some of thechemical
aureus, B. substilis and E. coli [19]. 0.2ml of known constituents found in these plants. The potential for
concentration 100µg (What was that conc.??) of developing antimicrobials from higher plants appears
inoculums was spread on the surface of the medium. Cups rewarding as it will lead to thedevelopment of a
(cavities) were made by using sterile borer (4mm) and phytomedicine to act against microbes.Plant-based
0.2ml of ciprofloxacin was poured into the cups of agar antimicrobials have enormous therapeutic potential as
showed maximum antimicrobial activity against Gram-
the antimicrobial effects of plant species, due to
Table 1: Phytochemical screening of leaves extract of C. halicacabum linn
S. No Chemical test Aqueous extract Alcoholic extract
1 Alkaloids + +
2 Carbohydrates + +
3 Fixed oils and fats - -
4 Saponins + +
5 Tannis and phenolic compounds - -
6 Proteins and free amino acids + +
7 Gums and mucilage - -
8 Flavinoids - -
9 Lignin + +
10 Phytosterol + +
11 Glycosides - -

0
5
10
15
20
25
30
E.Coli S.aureus B.substilis
Treatement
Zone of inhibition (mm)
50mg/ml 75mg/ml 100mg/ml standard
0
10
20
30
E.Coli
S.aureus
B.substilis
Zone of inhibition (mm)
Treatment
50mg/ml
75mg/ml
100mg/ml
standard
Advan. Biol. Res., 6 (1): 14-18, 2012
16
Table 2: zone of inhibition
Zone of inhibition(mm)
-----------------------------------------------------------------------------------------------------------------------------
Cardiospermum helicacabum linn. S. aureus, B. substilis E. coli
Aqueous 50µg 16 17 20
75µg 20 19 22
100µg 24 21 24
Alcoholic 50 µg 15 12 15
75 µg 17 15 17
100µg 19 18 19
Fig. 1: Zone of inhibition of various concentrations of aqueous extract of C. halicacabum linn
Fig. 2: Zone of inhibition of various concentration of alcoholic extract of C. halicacabum linn.
they can serve the purpose with lesser sideeffects that are aqueous extract compared to alcoholic (Fig 2 and 3). In
often associated with synthetic antimicrobials (Iwu et al., conclusion, ( Cardiospermum helicacabum linn…) broad
1999). However, the present study of antimicrobial spectrum of activity against a panel of bacteria
evaluation shows zone of inhihbition was more in responsible for the most common bacteria.

Advan. Biol. Res., 6 (1): 14-18, 2012
17
Extraction Procedure: 25 g of air-dried powder was taken 5. Alma, M.H., A. Mavi, A. Yildirim, M. Digrak and
in 200 ml of methanol in a conical flask, plugged with T. Hirata, 1725-1729. Screening chemical composition
cotton wool and then kept on a rotary shaker at 190-220 and In vitro antioxidant and antimicrobial activities
rpm for 24 h. After 24 hours the supernatant was collected of the essential oils from Origanum syriacum L.
and the solvent was evaporated to make the final volume growing in Turkey. Biol. Pharm. Bull., pp: 26.
one fourth of the original volume (12) and stored at 4 ¡C in 6. Klausmeyer, P., G.N. Chmurny, T.G. McCloud,
airtight bottles. K.D. Tucker and R.H. Shoemaker, 1732-1735.
Phytochemical Screening: Preliminary phytochemical Aniba panurensis. J. Nat Prod., pp: 67.
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for Wagneros), carbohydrates (+ve test result for Antimicrobial and cytotoxic activity of 18 prenylated
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result for Biuret test) and saponins. The other secondary Phytomedicine, 11: 666-672.
metabolites like steroids, cardiac glycosides, etc. were 8. Lin, F., M. Hasegawa and O. Kodama,
present in trace amounts in the extracts (Table 1). Purification and identification of antimicrobial
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than that of the standard drug (50mg/ml). But it was too 9. El-Seedi, H.R., N. Sata, K.B. Torssell and
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