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Research Article
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Cytotoxicity and Antifungal Activities of Ethanolic and Chloroform Extracts of Cucumis sativus Linn (Cucurbitaceae) Leaves and Stems |
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Joysree Das,
Anusua Chowdhury,
Subrata Kumar Biswas,
Utpal Kumar Karmakar,
Syeda Ridita Sharif,
Sheikh Zahir Raihan
and
Md Abdul Muhit
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ABSTRACT
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Cucumis sativus Linn. (Family: Cucurbitaceae) is commonly known as cucumber which has medicinal use in the entire world. The main objective of the present study was to examine the ethanolic and chloroform extracts of leaves and stems of the medicinal plant for cytotoxicity, antifungal activities and phytoconstituents. To determine the cytotoxicity and antifungal activities, brine shrimp lethality bioassay and agar disc diffusion methods were used, respectively. The extracts were subjected to in vitro cytotoxicity studies at 5, 25, 50, 100, 125, 150, 200, 250, 300, 350 and 400 μg mL-1 and antifungal study at the dose of 80 μg disc-1. The antifungal activity of the extracts was compared with standard drug, Griseofulvin at 30 μg disc-1. In brine shrimp lethality bioassay, the LC50 (μg mL-1) and LC90 (μg mL-1) of the ethanolic extract of Cucumis sativus were 35.48 and 141.25 μg mL-1, respectively. The chloroform extract of the cucumber also showed lethality against the brine shrimp nauplii (LC50: 75.86 and LC90: 151.36 μg mL-1). Moreover, the ethanolic extract and chloroform extracts of Cucumis sativus showed moderate antifungal activities against all tested organisms used in this study with zones of inhibition ranging from 4.40±0.18 to 1.67±0.08 mm and 3.45±0.04 to 1.50±0.12 mm, respectively. Thus, the ethanolic extracts of Cucumis sativus showed more potent cytotoxicity and Aspergillus niger was the most susceptible fungal strain to ethanolic extract of Cucumis sativus. The presence of different phytoconstituents e.g., alkaloid, glycoside, steroid, saponin, flavonoid and tannin as confirmed by preliminary phytochemical screening suggested that the identified compounds might play an important role for cytotoxicity and antifungal effects. Finally, it was concluded that ethanolic and chloroform extracts of leaves and stems of Cucumis sativus possessed cytotoxicity and antifungal activities.
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How
to cite this article:
Joysree Das, Anusua Chowdhury, Subrata Kumar Biswas, Utpal Kumar Karmakar, Syeda Ridita Sharif, Sheikh Zahir Raihan and Md Abdul Muhit, 2012. Cytotoxicity and Antifungal Activities of Ethanolic and Chloroform Extracts of Cucumis sativus Linn (Cucurbitaceae) Leaves and Stems. Research Journal of Phytochemistry, 6: 25-30.
URL: https://scialert.net/abstract/?doi=rjphyto.2012.25.30
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Received: October 14, 2011;
Accepted: October 21, 2011;
Published: February 13, 2012
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INTRODUCTION
Cucumis sativus Linn. (Family: Cucurbitaceae) is widely distributed
all over the world particularly in Asia, Africa and South America (Minaiyan
et al., 2011). Acylated flavone C-glycosides such as isovitexin
2-O-(6-(E)-p-coumaroyl) glucoside, isovitexin 2-O-(6-(E)-p-coumaroyl)glucoside-4-O-glucoside,
isovitexin 2-O-(6-(E)-feruloyl) glucoside-4-O-glucoside
and isoscoparin 2-O-(6-(E)-p-coumaroyl) glucoside were identified
from the leaves of Cucumis sativus plants (Abou-Zaid
et al., 2001). The ethanolic extract of the seeds of the plant showed
a significant activity against tapeworms and the results were comparable to
the effect of piperazine citrate (Elisha et al.,
1987). The ethanol extracts of powder fruit of Cucumis sativus produced
significant antidiabetic and hyperlipidemic effects in streptozotocin induced
rats (Karthiyayini et al., 2009). The hydroalcoholic
and buthanolic extracts of Cucumis sativus seeds were also effective
on controlling the loss of body weight in diabetic rats compared to controls
(Minaiyan et al., 2011). Moreover, sphingolipids
isolated from the crude methanol extract of cucumber (Cucumis sativus)
stems showed the antibacterial activity (Tang et al.,
2010). The crude extracts of Cucumis sativus fruits possessed moisturizing
property due to the presence of cucurbitacins (Prashant
et al., 2005). Abiodun and Adeleke (2010)
reported that the seeds of the plant served as good sources of proteins, fat
and minerals and the calcium content in the seeds of Cucumis sativus
was estimated to 2.03%. In addition to this, the preliminary phytochemical screening
of the aqueous extract of Cucumis sativus fruits showed the presence
of flavonoids and tannins which were responsible for antioxidant and analgesic
activities (Kumar et al., 2010). In the present
study, the cytotoxic and antifungal activities of the ethanolic and chloroform
extracts of Cucumis sativus leaves and stems were investigated. A preliminary
phytochemical screening was also carried out to identify the presence of various
types of phytoconstituents present in the extracts.
MATERIALS AND METHODS
Collection of plant materials: The leaves and stems of Cucumis sativus
were collected from Chandanaish in Chittagong of Bangladesh in the month of
July 2011 at day time. Fresh leaves and stems were cleaned with running tap
water and dried under the sun light. Then the plant parts were cut into small
pieces and homogenized to fine powder and preserved in airtight containers.
Preparation of plant extracts: The powdered materials were macerated with ethanol. Then the mixture was filtered and the filtrate was evaporated under reduced pressure in a rotary evaporator to obtain the crude ethanolic extract of Cucumis sativus. The same method was also employed to give the crude chloroform extract of the plant using chloroform as a solvent. Fungal strains: The antifungal activity of the plant extracts were investigated against six fungal strains such as Aspergillus niger, Blastomyces dermatitides, Candida albicans, Pityrosporum ovale, Trichophyton spp. and Microsporum spp. All the fungal strains were collected from Bangladesh Council of Scientific and Industrial Research (BCSIR), Chittagong, Bangladesh.
Phytochemical screening: Qualitative phytochemical analysis of the ethanolic
and chloroform extracts of the leaves and stems of Cucumis sativus was
carried out to identify the presence of various phytoconstituents (Trease
and Evans, 1989).
Cytotoxicity study and antifungal assay: The cytotoxic activities of
the ethanolic and chloroform extracts of Cucumis sativus were carried
out by brine shrimp lethality bioassay (Meyer et al.,
1982). On the other hand, agar disc diffusion method was used for antifungal
test of the crude extracts where potato dextrose agar medium was used to perform
the antifungal activity (Bauer et al., 1966).
Their antifungal activities were investigated against six fungal strains and
the results were compared with the standard drug, Griseofulvin (30 μg disc-1).
Statistical analysis: Three replicates of each sample were used for statistical analysis and the values were reported as Mean±SD (Standard Deviation). Online calculator was also used for the calculation of standard deviation.
RESULTS
Preliminary phytochemical analysis: Ethanolic extract of the leaves
and stems of Cucumis sativus showed the presence of alkaloid, glycoside,
steroid, saponin and tannin. Gum, flavonoid and reducing sugars were not found
in ethanolic extract of Cucumis sativus. On the other hand the chloroform
extract of the plant revealed the presence of alkaloid, glycoside, steroid,
flavonoid, saponin and tannin except gum and reducing sugars. Table
1 showed the results of phytochemical analysis of the ethanolic and chloroform
extracts of Cucumis sativus.
Cytotoxic assay: The cytotoxicity activity of the ethanolic and chloroform
extracts obtained from the plant against the brine shrimp nauplii under brine
shrimp lethality bioassay was shown in Table 2 and 3.
Table 1: |
Results of phytochemical analysis of the ethanolic and chloroform
extracts of Cucumis sativus |
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+: Indicates the presence and -: Indicates the absence of
the phytoconstituents |
Table 3: |
Brine shrimp lethality bioassay of chloroform extract of
Cucumis sativus |
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Table 4: |
Screening of the ethanolic and chloroform extract of Cucumis
sativus for antifungal activity |
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Data were represented as Mean±SD of triplicate determination.
-: No inhibition, SD: Standard deviation |
The LC50 (μg mL-1) and LC90 (μg
mL-1) of the ethanolic and chloroform extracts of Cucumis sativus
were deduced respectively (LC50: 35.48 and LC90: 141.25
μg mL-1; LC50: 75.86 and LC90: 151.36
μg mL-1). The best-fit line slope was used for calculation of
LC50 (μg mL-1) and LC90 (μg mL-1).
Antifungal assay: The results of antifungal screening assay of the ethanolic and chloroform extracts of the leaves and stems of Cucumis sativus against the tested strains were shown in Table 4. The ethanolic extract of Cucumis sativus showed the antifungal activity with zone of inhibition ranging from 4.40±0.18 to 1.67±0.08 mm. On the other hand, antifungal activity of chloroform extract of the plant was also observed with moderate zone of inhibition ranging from 3.45±0.04 to 1.50±0.12 mm. Finally, it was concluded that the crude ethanolic extract showed more antifungal activity than that of chloroform extract. DISCUSSION
No report was published earlier on the cytotoxicity, antifungal potentials
and phytochemical screening of the ethanolic and chloroform extracts of leaves
and stems of Cucumis sativus. The present study is the first report about
the cytotoxicity, antifungal potentials and phytochemical screening of the ethanolic
and chloroform extracts of leaves and stems of Cucumis sativus. In the
present study, the ethanolic extract of leaves and stems of Cucumis sativus
showed moderate antifungal action against all tested organisms used in this
study. The antifungal activity profile of the plant extracts against all tested
strains indicated that Aspergillus niger was the most susceptible fungus
while Microsporum spp. was the least sensitive strain of all the tested
fungus in this study. This indicated that both ethanolic and chloroform extracts
of leaves and stems of the medicinal plant have antifungal components. The preliminary
phytochemical screening of the ethanolic extract of leaves and stems Cucumis
sativus possessed phytoconstituents such as alkaloid, glycoside, steroid,
saponin and tannin except gum, flavonoid and reducing sugars while alkaloid,
glycoside, steroid, flavonoid, saponin and tannin were found in the crude chloroform
extract. Kumar et al. (2010) reported that the
aqueous extract of Cucumis sativus fruits revealed the presence of glycosides,
steroids, flavonoids, carbohydrates and tannins. Alkaloids and saponins were
not identified in the aqueous extract of fruits of the plant. The most common
phytochemicals found among the ethanolic, chloroform extracts of leaves and
stems and aqueous extract of fruits of the plant were glycoside, steroid and
flavonoid. The present study also confirmed the cytotoxic potentials of the
plant. LC50 values were 35.48 and 75.86 μg mL-1 while
LC90 values were 141.25 and 151.36 μg mL-1 of the
ethanolic and chloroform extracts of the plant, respectively (Table
3). Thus, it was concluded that the more potent cytotoxicity was exhibited
by the ethanolic extracts of leaves and stems of Cucumis sativus.
CONCLUSION It was concluded that the ethanolic and chloroform extracts of Cucumis sativus leaves and stems had cytotoxic and antifungal activities. These results claimed further investigation to isolate the main phytochemical compounds. The present study of in vitro cytotoxicity and antifungal evaluation of the plant was a preliminary investigation for future research work. So, further phytochemical and pharmacological studies on Cucumis sativus are strongly recommended to elucidate the exact chemical compounds and mechanisms involved. ACKNOWLEDGMENT Authors are thankful to the authority of BGC Trust University Bangladesh for providing the valuable support for the research work.
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