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International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395 -0056

Volume: 03 Issue: 03 | Mar-2016 www.irjet.net p-ISSN: 2395-0072

Screening of Potential Antimicrobial Activity of Indian Medicinal Plant of


Different Solvent Extract : Tinospora cordifolia and Hymenocallis
littoralis
Gyanendra Singh1, Rishi Kumar Saxena2 and Nishtha K Singh3
Mewar University, NH - 79, Gangrar, Chittorgarh, Rajasthan 312901
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Abstract - The present study reports that the screening of with sub terminal style scars, scarlet or orange coloured.
antimicrobial activity of Tinospora cordifolia and The fruit is pea-sized, subglobose drupe and red colored
Hymenocallis littoralis, medicinal plants extracts with on maturity. [1],[2],[3]
various different plant parts such as bulb, root, leaves and
stem against the opportunistic organisms such as Hymenocallis littoralis is a bulbous perennial herb of the
Staphylococcus aureus, Escherichia coli, Pseudomonas family Amaryllidaceae. It ranges in height from 60-70cm
aeruginosa, Salmonella typhimurium and fungi Candida (36 inches). It may be grown aquatically. They need
albicans using agar well diffusion method and Minimum average room humidity and temperature 70-85oF, 21-
inhibitory concentration (MIC). Antimicrobial efficiency of 29C during active growth; minimum winter temperature
Tinospora cordifolia and Hymenocallis littoralis, medicinal of 60/16C. Bright indirect sunlight is required for better
plants were examined using Ethanol, Methanol, Chloroform, growth with enough water content just enough to prevent
Dichloromethane, ethyl acetate as solvents a nd found that the soil from drying out completely. It is commonly
all the plants parts showed significant activity against all employed as an ornamental plant, promoting blood
pathogens. circulation; antineoplastic, cytotoxicity and antiviral.,
antioxidant & anticancer activities. Leaves are 10-12,
The best MIC (zone of inhibition in mm) results were found sessile; lamina sword shaped, 45-75cm long, 2.5-6cm
to be 20mm in case of Staphylococcus aureus by using stem wide, tip suddenly sharpened, basal part gradually
crude extract followed by the Escherichia coli narrowed, deep green, with many veins. Flower are white,
(19mm),Salmonella typhimurium (19mm), Pseudomonas large and sessile; perianth tube cylindrical, slim, varied in
aeruginosa (17mm) and Candida albicans (16mm). Stems length, longer individual up to more 10cm long. The bulbs
extracts from the H. littoralis against Escherichia coli of Hymenocallis is spherica 7-10cm (3-4 inches) in
followed Staphylococcus aureus, Pseudomonas aeruginosa, diameter.[4],[5],[6]
Salmonella typhimurium, Candida albicans produced
highest zone of inhibition at 24mm followed by 21mm, 18 The present study focused on the screening of
mm, 17mm and 17mm respectively. antimicrobial activities of the Indian medical plants such
as Tinospora cordifolia and Hymenocallis littoralis by using
The Spectrum of activity observed in the present study may different solvents for the purpose of extraction on the
be indicative of the present study methanol extracts of these different parts of these respective plants.
plants could be a possible source to obtain new and effective
herbal medicines to treat different diseases or disorders. 2. MATERIAL AND METHODS
Key Words - Tinospora cordifolia, Hymenocallis
1.1 Collection of Plant Materials
littoralis, Antimicrobial, MIC, Agar Well Diffusion.
Bulb, Stem, leaves and flowers of Tinospora cordifolia and
1. INTRODUCTION Hymenocallis littoralis used in this study were collected
Amruthvalli / Guduchi, Tinospora cordifolia, known by the from mumbai region.
common names Heart-leaved Moonseed, Guduchi and
1.2 Plant Extract Preparation
Giloy, is an herbaceous vine of the family Menispermaceae.
Many interesting findings in the areas of immuno Each plant parts (Bulb, Stem, leaves and flowers) of
modulation, anticancer activity, liver disorders, Hymenocallis littoralis (Jacq.) Salisb were carefully cut and
antidiarrhoeal, anti-oxidantactivity, aphrodisiac activity, washed with running tap water and then with clean sterile
anthelmintic activity, antipsychotic activity and water to remove dirt prior to the drying process. Each of
hypoglycemic are reported. Leaves are simple, alternate, the plant parts were cut into small pieces and dried at
exstipulate, long petioles up to 15cm long, bears heart- 45C for 48 hours to remove the moisture content. Finally,
shaped, whitish to mentose with prominent reticulum explants was ground using mortar and pestle into fine
beneath. Flowers are unisexual, small on separate plants. powder sample.
Male flowers clustered, female usually solitary. Fruits are
aggregate of 1-3, ovoid smooth drupelets on thick stalk
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International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395 -0056
Volume: 03 Issue: 03 | Mar-2016 www.irjet.net p-ISSN: 2395-0072

1.6 Minimum Inhibitory concentration


1.3 Solvent Extractions Minimum Inhibitory concentration of different extracts
were evaluated modified by [9]. The minimum inhibitory
The powdered plant materials were extracted using concentration is defined as the lowest concentration able
different solvents Ethanol, Methanol, Chloroform, to inhibit any visible bacterial growth on the culture plates
Dichloromethane, Ethyl acetate by maceration technique or in the tubes based on turbidity. This was determined
to obtain the crude extracts. 1 gram of dried samples was from readings on the culture plates after incubation. The
ground using pestle and mortar, before adding 10 ml of most commonly employed methods are the tube dilution
methanol. The homogenate was filtered through four method and agar dilution methods. Serial dilutions are
layers of miracloth and centrifuged at 4700 rpm for 5min made of the products in bacterial and fungal growth
at 25C. Supernatent was taken for further studies or were media. The test organisms are then added to the dilutions
stored under refrigeration (-20C) condition for further of the products, incubated, and scored for growth. MIC is
analysis. generally regarded as the most basic laboratory
measurement of the activity of an antimicrobial agent
1.4 Test Microorganisms against an organisms. The antimicrobials present in the
plant extract are allowed to diffuse out into the medium
Total 4 bacteria in which Gram positive bacteria, namely and interact in a plate freshly seeded with the test
Staphylococcus aureus and Gram negative bacteria namely organisms. The resulting zones of inhibition will be
Escherichia coli, Pseudomonas aeruginosa, Salmonella uniformly circular as there will be a confluent lawn of
typhimurium and fungal pathogen such as Candida growth. The diameter of zone of inhibition can be
albicans were used from our laboratory. All bacteria were measured in millimeters.
maintained on Nutrient Agar (NA) at 37C and fungi on
Potato Dextrose Agar (PDA) & Saborauds dextrose agar
(SDA) at 28C. 3. RESULT AND DISCUSSION

1.5 Agar Well Diffusion Assay For a long period of time, plants have been a valuable
source of natural products for maintaining human health,
Antimicrobial activities of different extracts were especially in the natural therapies. In the present study,
evaluated by the agar well diffusion method [7] modified five solvents extracts viz Ethanol, Methanol, Chloroform,
by [8]. Nutrient agar (NA) and Sabourauds Dextrose Agar Dichloromethane, Ethyl acetate were taken for
(SDA) plates were poured innoculated with 24 hrs and 48 antimicrobial activity by agar well diffusion method where
hrs old broth culture of bacterias and fungi respectively in comparision with other solvents, Methanol crude
under aseptic conditions. The plates were covered and extract of Tinospora cordifolia showed best activity against
allowed to cool. As soon as the agar was partly solidified, Escherichia coli, Staphylococcus aureus, Pseudomonas
the plates were inverted and left for 2 hrs. Then, wells aeruginosa, Salmonella typhimurium and Candida albicans.
were made at the centre of the plate by using a 6 mm cork
borer that was sterilized with alcohol and flame. Stock The best MIC (zone of inhibition in mm) results were
solution of each plant parts extract was prepared at a found to be 20 mm in case of Staphylococcus aureus by
concentration of 1 mg/10ml in methanol. About 100 l of using stem crude extract followed by the Escherichia coli
different concentrations of plant solvent extracts were (19mm), Salmonella typhimurium (19mm), Pseudomonas
added by sterile micropipettes into the wells and allowed aeruginosa (17mm) and Candida albicans (16mm). Stem
to diffuse at room temperature for 2 hrs. The 5 solvents crude extract were shown best activity against Escherichia
viz., ethanol, methanol, chloroform, dichloromethane, coli, Staphylococcus aureus, Pseudomonas aeruginosa
ethyl acetate at different volumes were used as control Salmonella typhimurium and Candida albican are followed
whereas Gentamycin and Caspo at same concentration by roots, leaves and bulbs crude extract of Tinospora
with plant extract was used as the reference. The plates cordifolia are given in Table 1.
were incubated at 37C for 18-24 hrs for bacterial
pathogens and 28C for 48 hours fungal pathogens.
Diameter of the inhibition zone (mm) was measured, the
readings were taken in three different fixed directions and
the average values were recorded.

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International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395 -0056
Volume: 03 Issue: 03 | Mar-2016 www.irjet.net p-ISSN: 2395-0072

Table 1: Antimicrobial activity of Methanol crude Khan, 2012 [12] studied that water, ethanolic and ethyl
extract of Tinospora cordifolia. acetate extracts of Tinospora average inhibitory zone of
14mm, 17mm and 12.5mm respectively which indicates
that ethanolic extract shows best result having zone of
inhibition more than that of tetracycline (12mm against all
pathogens). In case of Tinospora stem and root best result
was shown by hot aqueous extract with a zone of
inhibition of 16mm and 17mm respectively.

Five different Solvents samples and their crude extracts


(Ethanol, Methanol, Chloroform, Dichloromethane, Ethyl
acetate) of different parts such as Roots, stem, leaves and
bulbs from Hymenocallis littoralis were used to screen the
antimicrobial activity, Meanwhile, five microbes were used
to test all the samples, namely Gram positive bacteria
(Staphylococcus aureus), Gram negative bacteria
(Salmonella typhiimurium, Pseudomonas aeruginosa,
Escherichia coli and fungal strain (Candida albicans).

Overall, Stem crude extract showed pronounced inhibition


of microbes growth of the cultures compared to Roots,
Leaves and bulbs crude extract with different range on
inhibition zone (Table 2). Most probably, the highest
amount of secondary metabolite was presented such as an
alkaloid in mature plant was the main reason for these
results obtained.

Table 2: Antimicrobial activity of Methanol crude extract


of Hymenocallis littoralis.

Fig. 1 : Well Agar Diffusion Assay of Methanol Extract of


Tinospora cordifolia

Similar studies in the past have documented by Kumari,


2012 [10] that the antimicrobial activity of different
extracts of Tinospora cordifolia in various micro-
organisms. Methanolic extract of Tinospora cordifolia was
found to have antimicrobial activity against E coli
(MTCC1), Staphylococcus aureus (MTCC98), Salmonella
typhimurium (MTCC737). The largest zone of inhibition
(18 mm) was found against Staphylococcus aureus and the
least activity was observed for Salmonella typhimurium (8
mm).

In another study by Singh et al (2012) [11] have concluded


that maximum antibacterial activity of hot and cold
methanol extracts was exhibited against Staphylococcus
aureus when compared with standard drug Ciprofloxacin
19. In case of Staphylococcus aureus for methanolic extract
the zone of inhibition was 20mm, 14mm and 18mm for
1gm/ml, 500mg/ml, 250mg/ml respectively followed by
Staphylococcus aureus, Pseudomoas aeruginosa and then
Escherichia coli.

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International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395 -0056
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4. CONCLUSION

Different parts of the plant such as Roots, stem, leaves and


bulbs from Hymenocallis littoralis showed pronounced
inhibition of microbes growth. Stems of the Hymenocallis
littoralis as well as Tinospora cordifolia extracts produced
highest inhibition activity against Staphylococcus aureus
and E. coli, followed by the Roots, leaves and bulbs
respectively against Pseudomonas aeruginosa, Salmonella
typhiimurium, Candida albicans . Present study proves that
the methanolic extract of different parts of Tinospora
Fig. 2 : Well Agar Diffusion Assay of Methanol Extract of cordifolia and Hymenocallis littoralis can be used for
Hymenocallis littoralis medicinal purposes. Revealed that T. cordifolia and H.
littoralis is an excellent drug, which could be a good
Stems extracts from the H. littoralis against Escherichia coli remedy for various ailments of animals as well as human
followed Staphylococcus aureus, Pseudomonas aeruginosa, beings.
Salmonella typhiimurium, Candida albicans. produced
highest zone of inhibition at 24mm followed by 21mm, 18 REFERENCES
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International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395 -0056
Volume: 03 Issue: 03 | Mar-2016 www.irjet.net p-ISSN: 2395-0072

[11] Singh S. and Singh P., Effectiveness of Tinospora


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