Investigation of Antimicrobial activity and Chemical Constituents of Eragrostis cynosuroides by GC-MS

 

M. Barath¹, J. Aravind¹, R. Sivasamy²*

¹Dept. of Biotechnology, Rajalakshmi Engineering College, Chennai, Tamilnadu, India.

²Dept. of Human Genetics and Molecular Biology, Bharathiar University, Coimbatore, Tamilnadu, India.

 

ABSTRACT:

Medicinal plants are rich in various phytoconstituents which make them to be a potential candidate for curing diseases. Even though there are many medicines which are chemically synthesized now are available, these when consumed by the patient cause many adverse effects other than curing the disease. These effects are mostly incurable. The patient suffers by the disease and also by the side effects of the drug. But the traditional way of treating disease in most of the countries is by using various herbal plants. These plants have more chemical compounds which have the ability to cure the disease. When the extract of the plant is applied over the diseased surface or when consumed by the patient, the disease gets cured without causing any side effects. Eragrostis cynosuroides known as (Dharbham -in Sanskrit) is a medicinal plant belonging to family Poaceae, most prominently used in Asian countries. This Plant has an extensive property of acting as anticancerous, antimicrobial and antioxidant agents. And this plant has an ability to absorb ultraviolent radiation as well. These Medicinally useful part of the plants were subjected to analyze Phytochemical properties by GC-MS. The GC-MS results revealed that, compounds like Docosanoic acid, Eicosyne, Eicosenoic acid and Phytol etc., which were obtained, play an important role in the above mentioned antimicrobial activity.

 

INTRODUCTION:

Medicinal plants play a key role in the human health care. Most of the world populations depend on the role of traditional medicine uses plant materials to treat the ailments. The traditional medicine like ayurveda, siddha, amchi and unani always plays a major role in treating diseases. There are many herbs which contain various constituents which make the herb a potable candidate in therapeutic process^1,2. These medical practices originated a long time ago and developed gradually by practical methods without any modern principles. Since very old times, herbal medications have been used for relief of symptoms of disease. Even though there are many chemically synthesized drugs available, plants always plays an important role in health care.

 

The plant Eragrostis cynosuroides is a herbal plant which has many medicinal activities like anti-asthmatic, anti-inflammatory, antipyretic, galactagogue, anti-diuretic, astringent, aphrodisiac, sedative etc. this plant along with other herbs are used to cure, nausea, menorrhagia, jaundice, skin infections etc^3,4,5. This plant is a salt and drought tolerant plant which has deep rhizome that helps to bind tightly with the sand⁶. With this background information, the present study aimed to investigation of Antimicrobial activity and Chemical Constituents of Eragrostis cynosuroides

 

METHODS:

Plant Material:

The Plant Material was collected from Palar river bank, Chengalpattu District, Tamil Nadu, India. The plant material was botanically identified as Eragrostis cynosuroides. There are many synonyms for this plant like, Desmosta chyabipinnata, Eragrostis bipinnata L., Brizabipinnata L. Eragrostisis genus of the Poaceae (grass family) and the type genus of the tribe Eragrostideae commonly known as cane grass. or love grass.

 

Extract preparation:

The leaf of Eragrostis cynosuroides (5 kg) was collected and shade dried ground to fine powder and used for the extraction. The compounds were extracted using three different solvent systems. Briefly, each 25g of dried sample were extracted twice with 500 ml of Acetone (Polar aprotic Solvent), Chloroform (Non-polar solvent) and 95% methanol (Polar protic solvent) at 25°C for 48 h the crude extracts was filtered through with what man No 1 Paper and concentrated using a rotary evaporator under reduced pressure at 40⁰C. The crude compounds were dissolved with DMSO and used for the further analysis.

 

GC-MS Analysis:

The crude extracts of Acetone, Chloroform and Methanol from Eragrostis cynosuroides were subjected to GC-MS analysis. GC-MS analysis was performed with JEOL GCMATE II GC-MS instrument with high resolution 6000 maximum, double focusing instrument with maximum calibrated mass 1500 Daltons. Result prediction on mass-spectrum GC-MS was obtained using the NIST (National Institute Standard and Technology) database. The range of the known components was compared with the range of the unknown components present in the NIST library. The structure, name and molecular weight of the components of the test materials were verified^7,8.

 

Antimicrobial Activity:

Antimicrobial activity is where the plant extract used against various strains of microorganisms which infects the humans in many ways and cause disease. Reports on the antimicrobial activity of the Eragrostis cynosuroides shows that the root extract of this plant show great effect against the growth of bacteria like Staphylococcus aureus Escherichia coli, Klebsiella pneumonia⁹. To determine the antibacterial activity of Eragrostis cynosuroides we used the methanol and chloroform (1:2) extract by disc diffusion method¹⁰. The bacterial strains like E. coli, and Bacillus subtilis were used to for testing antibacterial activity. Streptomycin (10 μg ml 1) used as positive control and DMSO solvent used as negative control.

 

The zone of inhibition is measured in mm. Fungal strains like Aspergillus fumigates, Candida albicans, Candida tropicalis, Pencillium chrysogenum and Aspergillus flavus were tested with the plant extract to determine the antifungal activity of the extract¹¹. In this investigation we used the fungal strain of Aspergillus niger to test the antifungal activity of the plant extract. Extract of the plant has been impregnated into a disc and is used in plates containing the fungi. Blank disc impregnated with the solvent followed by drying off was used as negative control and Nystatin (10 μg disc 1) used as positive control. Then the zone of inhibition obtained is measured in mm.

 

RESULTS AND INTERPRETATION:

Identification of Phytocomponents in DH001:

GC-MS chromatogram analysis of the acetone extract of Eragrostis cynosuroides (figure 1) showed nine peaks which indicating the presence of nine Phytochemical constituents. on comparison of the mass spectra of the constituents with the NIST library, the nine compounds were characterized and identified [Table 1].

 

 

Figure 1. DH001- Acetone extract

Table 1- source: GC-MS / Molecular formula - Pubchem. ncbi.

 

Identification of Phytocomponents in DH002:

GC-MS chromatogram analysis of the chloroform extract of E. cynosuroides (figure 2) showed ten peaks. Each peak represents one chemical constituent. So ten peaks represent ten compounds. On comparing the mass spectra of the constituents with the NIST library, the ten compounds were characterized and identified [Table 2].

 

 

igure 2. DH002- chloroform extract

 

Table 2- Source: GC-MS / Molecular formula - Pubchem.ncbi

 

 

Identification of Phytocomponents in DH003:

GC-MS chromatogram analysis of the methanol extract of E. cynosuroides (figure 3) showed twelve peaks which indicating the presence of twelve Phytochemical constituents. On comparison of the mass spectra of the constituents with the NIST library, the twelve compounds were characterized and identified [Table 3].

 

Figure 3. DH003- methanol extract

 

Table 3- Source: GC-MS / Molecular formula - Pubchem.ncbi

 

Antimicrobial Activity:

The zone of inhibition obtained by using the discs impregnated with the plant extract is calculated to find the antimicrobial efficiency of the plant extract.  The antibacterial activity of the plant extract against E. coli and B. subtilis was shown in the figure 4 and figure 5. The antifungal activity was shown in the figure 6. And it’s corresponding value of inhibition zone was given in the table 4.

 

Figure 4: Antibacterial activity (E. coli)

NC-Negative Control, PC-Positive Control

 

PC

Extract

NC

Figure 5: Antibacterial activity (B.subtilis)

NC-Negative Control, PC-Positive Control

 

Figure 6: Antifungal activity (A. niger)

NC-Negative Control, PC-Positive Control

 

Table 4: Table showing the anti-microbial activity

Microbial culture	Zone of inhibition	Negative control	Positive control
E. coli	8 mm	0 mm	10 mm
B. subtilis	5 mm	0 mm	10 mm
Aspergillis niger	5 mm	0 mm	0 mm

 

DISCUSSION:

The plant extract of Eragrostis cynosuroides has been reported to have many therapeutic activities. So the methanol extract of the plant is obtained and checked for various phytoconstituents present in it and also the plant’s antimicrobial activity. The GC-MS has been used to determine all the phytoconstituents present in it and disc diffusion method is used to determine the antimicrobial activity of the plant extract. The result showed that the acetone extract yielded 9 compounds, the chloroform extract yielded 10 compounds and the methanol extract yielded 12 compounds. The antimicrobial activity which was tested using the disc diffusion method showed that the extract was very effective against the bacteria E. coli and against the fungi A. niger. B. Subtilis was also found to be affected by the plant extract as it showed inhibited growth but not much when compared to the others. By all the results obtained, the plant extract of Eragrostis cynosuroides has been proved to have various phytoconstituents with antimicrobial activity.

 

REFERENCE:

1.       Al-Kouthayri, G.R., and A. A. Hassan. Survey of major weeds in Hadramout Valley, Yemen. Arab Journal of Plant Protection 16(1); 1998: 19-26.

2.       Fowler, G. 2002. Distribution Map. USDA, APHIS, PPQ, Center for Plant Health Science and Technology, Raleigh, NC.

3.       Kirtikar, K.R., Basu, B.D. Indian Medicinal Plants. 4th Vol. Dehradun, India: International Book Distributors Book Sellers and Publishers. 1918.

4.       Joshi, S.G., 2003. Medicinal Plants. New Delhi, India: Oxford and IBH Publishing Co. Pvt Ltd., pp-318

5.       Alikhan, A. Khanun, A., 2004. Ethnomedicine and human welfare.1st ed. Hyderabad, India: Ukaaz Publication

6.       Ashish Pandey, Satish Kumar Sharma, Lalit Singh, Tanuja Singh. An overview on Desmostachyabippinata. Journal of Drug Discovery and Therapeutics1 (7); 2013: 67-68.

7.       Bagavathi Perumal Ezhilan, Ramasamy Neelamegam. GC-MS analysis of phytocomponents in the ethanol extract of Polygonumchinense L. Pharmacognosy Research 4(1); 2012: 11-14.

8.       Ramasamy Sivasamy, Jayaraman Angayarkanni and Muthuswamy Palaniswamy. A novel filarial topoisomerase II inhibitor produced by native isolate Micrococcus luteusB1252. African Journal of Biotechnology. 10(71); 2011: 16069-16077.

9.       Hashmi H and Rashid A. Isolation of fungi from roots of Parthenium hysterophorus and Desmostach yabipinnata and antibacterial activity of their root extracts. Journal of Biological Sciences 1: 2001; 350-351.

10.     Taylor R.S.L., N.P. Manandhar, J.B. Hudson and G.H.N. Towers. Screening of selected medicinal plants of Nepal for antimicrobial activities. Journal of Ethnopharmacology. 546; 1995: 153-159

11.   Zain ME, Awaad AS, Al Othman MR and Al-Dosary SK. Antibacterial, antifungal activity and Phytochemical analysis of some desert plants against human pathogenic bacteria and fungi. Life Science Journal 11(7);2014: 343-349.

 

 

 

Received on 02.02.2016          Modified on 14.02.2016

Accepted on 28.02.2015        © RJPT All right reserved