Phytochemical Investigation with Assessment of Cytotoxicity and Antibacterial Activities of Chloroform Extract of the Leaves of Kalanchoe pinnata
Kalanchoe pinnata is used in traditional medicines for the treatment of several health problems in Bangladesh. The study was designed to investigate the cytotoxicity and antibacterial activities of the chloroform extract of the leaves of Kalanchoe pinnata. Qualitative phytochemical analysis was also performed for the identification of several phytochemical compounds such as alkaloids, glycosides, steroids, gums, flavonoids, saponins, reducing sugar, tannins and terpenoids. The study revealed that the plant extract contained all of the phytoconstituents except steroids, flavonoids and terpenoids. The agar disc diffusion technique was used to investigate the antibacterial activities of the plant extract at 500 μg disc-1 against eight bacterial strains. The plant extract showed very low antibacterial activities with zone of inhibition ranging from 0.33±0.02 to 1.67±0.02 mm, while the maximum antibacterial action was recorded against E. coli with zone of inhibition of 1.67±0.02 mm but no sensitivity was found against V. cholerae. The antibacterial activity of the plant extract was compared with the standard drug, Levofloxacin which was used at 10 μg disc-1. Brine shrimp lethality bioassay was also done to find out the cytotoxic activity of the plant sample. The chloroform extract of the plant showed lethality against the brine shrimp nauplii (LC50: 125.89 and LC90: 234.42 μg mL-1. Finally, it was concluded that the chloroform extract of the plant leaves possessed considerable cytotoxic activity with less antibacterial potentials.
to cite this article:
S.K. Biswas, A. Chowdhury, S.Z. Raihan, M.A. Muhit, M.A. Akbar and R. Mowla, 2012. Phytochemical Investigation with Assessment of Cytotoxicity and Antibacterial Activities of Chloroform Extract of the Leaves of Kalanchoe pinnata. American Journal of Plant Physiology, 7: 41-46.
Received: November 04, 2011;
Accepted: January 24, 2012;
Published: February 22, 2012
Medicinal plants were used as the principal health care agent before the discovery
of modern medicines (Shafaei et al., 2011). The
plants and phytochemical compounds are used in folk medicine for the treatment
of different types of diseases (Pareta et al., 2011).
The medicinal plant Kalanchoe pinnata Linn belonging to the family of
Crassulaceae is found in Bangladesh and the local name of the plant is patharkuchi
(Ghani, 2003). Several pharmacological potentials such
as immunosuppressive (Rossi-Bergmann et al., 2006),
anti-inflammatory and analgesic (Joseph et al., 2011),
hepatoprotective (Yadav and Dixit, 2003) and antitumor
promoting activities (Supratman et al., 2001)
were reported. Inflammation of nephrons (Hartalka et
al., 2007), hypertension (Lans, 2006) and allergic
conditions (Cruz et al., 2008) can also be treated
with the plant extract.
The ethanol extract of the leaves of Kalanchoe pinnata extracts possessed
significant antibacterial activity (Biswas et al.,
2011) and the petroleum ether and aqueous extracts of the plant showed cytotoxicity
and antifungal potentials (Chowdhury et al., 2011).
The present study was designed to find out the cytotoxic properties and antibacterial
activities of the chloroform extract of the leaves of Kalanchoe pinnata.
The preliminary phytochemical screening of the plant extract was also done.
MATERIALS AND METHODS
Collection and preparation of plant samples: The leaves of the medicinal
plant were collected in December 2009 at day time from Ramchandrapur, Magura,
Bangladesh. The undesirable materials were separated from leaves and after that,
the leaves were air-dried for three weeks. The plant parts were ground into
fine powder and kept in an air-tight container until use. About 200 g of powder
of the plant material was soaked in 1000 mL of chloroform in a glass container
which was sealed and stored for three weeks followed by continuous shaking.
Finally, the crude mixture was filtered and the obtained filtrate was completely
evaporated to give a gummy concentrate as crude chloroform extract.
Preliminary phytochemical screening: The freshly prepared chloroform
extract of Kalanchoe pinnata was qualitatively tested for the presence
of phytochemical compounds such as alkaloids, glycosides, steroids, gums, flavonoids,
saponins, reducing sugar, tannins and terpenoids. The standard procedures as
preliminary screening (Trease and Evans, 1989) were used
to identify the presence of different phytoconstituents in the plant extract.
Test of antibacterial activity and cytotoxicity of the plant extract:
The cytotoxic activity of the crude extract was investigated by brine shrimp
lethality bioassay (Bauer et al., 1966) and agar
disc diffusion method (Meyer et al., 1982) was
used to determine the growth of inhibition of the bacterial strains.
Statistical analysis: Three replicates of each sample were used for statistical analysis and the results of the experiment were expressed as mean±standard deviation (SD).
Table 1 showed the results of phytochemical screening of the chloroform extract of the plant which revealed the presence of alkaloids, glycosides, gums, saponins, tannins and reducing sugars but the extract did not reveal the presence of steroids, flavonoids and terpenoids.
Cytotoxic activity: The results of investigation of cytotoxic activity (Table 2) showed that the chloroform extract of the plant possessed moderate level of general toxicity in the brine shrimp lethality bioassay (LC50: 125.89 and LC90: 234.42 μg mL-1).
Antibacterial activity: Antibacterial activity of the chloroform extract
was investigated against eight bacterial strains. The results of antibacterial
test showed that the crude extract showed very low level of antibacterial activity
against B. subtilis, B. megaterium, S. aureus, E. coli,
P. aeruginosa, S. typhi and S. dysenteriae with zone of
inhibition between 0.33±0.02 to 1.67±0.02 mm.
|| Results of phytochemical analysis of the chloroform extract
of the leaves of Kalanchoe pinnata
|+: Present and -: Absent
|| Brine shrimp lethality bioassay of the chloroform extract
of the leaves of Kalanchoe pinnata
|| Screening for antimicrobial activity of the chloroform extract
of the leaves of Kalanchoe pinnata
|Data were represented as Mean±SD of triplicate determination,
-: No inhibition
The highest zone of inhibition or antibacterial activity was found against
E. coli (1.67±0.02 mm) while no antibacterial activity was found
against V. cholerae. The zone of inhibition of the plant extract is shown
in Table 3.
Alkaloids, glycosides, gums, saponins and reducing sugars were identified in
the chloroform extract of the leaves of Kalanchoe pinnata while Chowdhury
et al. (2011) reported the presence of alkaloid, glycoside, gums,
saponins, reducing sugar and tannins in the petroleum ether extract of the leaves
of Kalanchoe pinnata. They also reported that the aqueous extract of
the same plant contained alkaloids, glycosides, steroids, saponins, tannins.
On the other hand, the ethanol extract of the medicinal plant possessed alkaloids,
glycosides, steroids, gums, flavonoids, saponins, reducing sugars and tannins
(Biswas et al., 2011). The reducing sugars, tannins
and glycosides present in the extract may be responsible for antibacterial activity
and cardiac stimulatory potentials (Sofowora, 1993; Hagerman
et al., 1998).
From literature review, it is evident that the phytochemical compounds play
an important role in the therapeutic effects of the plants such as flavonoids
for free radical scavenging potentials and antinociceptive activities (Zakaria
et al., 2006). Alkaloids and tannins have also significant effect
in inhibition of pain perception (Uche and Aprioku, 2008;
Rahman et al., 2011; Ramprasath
et al., 2006). Moreover, tannins could prevent infectious diseases
(Agbafor et al., 2011).
The obtained results also showed the cytotoxic activity against brine shrimp
nauplii with LC50: 125.89 and LC90: 234.42 μg mL-1.
In comparison with the cytotoxic activities of the extract of petroleum ether
(LC50: 25.12 and LC90: 177.83 μg mL-1)
and aqueous extract (LC50: 25.12 and LC90: 173.78 μg
mL-1) of the leaves of the plant reported by Chowdhury
et al. (2011) and that of ethanol extract (LC50: 100 and
LC90: 204.17 μg mL-1) by (Biswas
et al., 2011), the chloroform extract possessed less cytotoxic potentials.
In addition to cytotoxic properties, the chloroform extract showed very low
antibacterial activities showing zone of inhibition ranging from 0.33±0.02
to 1.67±0.02 mm while significant antibacterial activity was found in
the ethanol extract of the leaves of the plant with zone of inhibition between
6.0±0.35 to 8.2±0.22 mm (Biswas et al.,
The results obtained from this study suggested that the chloroform extract of the leaves of Kalanchoe pinnata might contain anticancer and antibacterial compounds. However, more research work is needed to isolate the active chemical compounds with their chemical structures elucidation.
The authors are grateful to the authority of BGC Trust University for extending their cordial support to perform this research work.
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