Mr.Vishal Tagalpallewar 1 *, A.S.Kediya
2 A.G.Moon
3 ,Dr.N.H.Indurwade
4
Dept. of Pharmaceutical Chemistry,Dr.R.G.Bhoyar Institute of Pharmaceutical Education &
Research,Wardha-.442001
1
Email id vishaltagalpallewar@gmail.com
ABSTRACT:
Lemon grass commonly known as Cymbopogon flexuousus , belonging to family Graminae distributed throughout the India. The plant is used to obtain lemon grass oil and it can be used as flavoring agent. It is employed as a source of citral from which β-ionine is prepared to synthesize vitamin A. The present study is aimed towards antibacterial and antioxidant activity of essential oil of Cymbopogon flexuousus . Cup plate agar diffusion method was used for determination of antibacterial activity. For determination of antioxidant activity Nitric oxide radical inhibiting activity and 2,2-diphenyl-1-pickryl hydrazyl (DPPH) method using ascorbic acid as standard was employed. The essential oil was tested against pathogenic strains of Gram positive ( Staphylococcus aureus ) and Gram negative ( Pseudomonas aeruginosa) against standard drug for determination of zone of inhibition. The essential oil exhibited a good antibacterial activity against gram positive and gram-negative organisms with maximum zone of inhibition of 26mm against S.aureus
. The essential oil also exhibits antioxidant activity by Nitric acid method and DPPH method. By nitric acid method the extract shows antioxidant activity with IC
50
value 121

8.25
 g/ml where as with
DPPH method the IC
50
value was found to be 602.17

5.11
 g/ml.
Keywords: β-ionine, DPPH, zone of inhibition, IC
50.
INTRODUCTION
Lemon grass commonly known as Cymbopogon flexuousus , belonging to the family Graminae is distributed throughout the India. The plant is used to obtain lemon grass oil and it can be used as

flavoring agent, antiseptic and deodorant. It is employed as a source of citral from which β-iodine is prepared to synthesize vitamin A1. The present study is aimed towards antibacterial and antioxidant activity of essential oil of Lemon grass leaves.

MATERIALS AND METHODS:

Collection of plant material:

The leaves of Cymbopogon flexuousus were collected from Wardha of Maharashtra.

Extraction of essential oil:
 The fresh leaves were cleaned, cut into small pieces and used for extraction of essential oil by hydro distillation method using Clevenger apparatus for 8 hours. The oil recovered was dried over anhydrous sodium sulphate and kept in the refrigerator at 4
0
C and used for further study
2
.

Nausea and vomiting are greatly avoided by buccal drug delivery.

Used in case of unconscious and less co-operative patients.
ANTIMICROBIAL STUDY:
Microorganisms:
Two pathological strains of Gram positive ( Staphylococcus aureus ATCC 25923 and Bacillus subtilis - MTCC 441) and Gram negative ( Escherichia coliATCC 25922 and Pseudomonas aeruginosa -ATCC 27853) organism were used for the antibacterial activity 3 . All the microorganisms were obtained from the laboratory stock of the Department of Pharmaceutical microbiology.
Media 4 :
Nutrient agar (13 gm in 1000 ml distilled water) was used in the study.
Antimicrobial agents:
Penicillin (100µl/ml) and streptomycin (100µl/ml) were used as standard for antibacterial activity of gram positive and gram-negative bacteria respectively.
Determination of antibacterial activity 5, 6 :
Cup plate agar diffusion method was used to determine zone of inhibition of essential oil of Lemon grass. The essential oil of concentration of 100 µl/ml was prepared by using Dimethyl sulfoxide
(DMSO). By using nutrient agar medium, inoculated plates was prepared in triplicate, in each plate

three bore of 6mm size was done for each essential oils, standards and solvent control (DMSO) for each organism. The plates were incubated at 37
0
C and determined the zone of inhibition after24 hours.
ANTIOXIDANT STUDY:
1.
Nitric oxide radical inhibiting activity 7 :
Method : The reaction mixture (6ml) containing Sodium nitroprusside (10 mm, 4 ml), Phosphate buffer solution (1 ml) & essential oil (1ml) prepared in DMSO (1000 μg/ml) was incubated at 25 0 C for 180 min. After incubation, 0.5 ml of reaction mixture containing nitrate was removed, 1 ml of
Sulphanilic acid reagent (0.33 % in 20 % Glacial acetic acid) was added, mixed well & allowed for stand for 5 min for completion of diazotisation then 1 ml of Napthyl ethylene diamine dihydrochloride (NEDD) was added, mixed & allowed to stand for 30 min in diffused light. A pink colored chromophore was formed. The absorbance of these solutions was measured at 540 nm against corresponding blank solution. IC50 value is the concentration of the samples required to inhibit 50% nitric oxide radical.
2.
Diphenyl picryl hydrazyl (DPPH) method 8 :
Method:
The assay was carried out in 96 well microtitre plate. To 200 μl of DPPH solution, 10 μl of each test sample or the standard solution was added separately in wells of microtitre plate. The final concentration of the test & the standard solution used are 1000 μg/ml, 500μg/ml, 250 μg/ml,
125 μg/ml, 62.5 μg/ml, 31.25 μg/ml & 15.625 μg/ml. The plates were incubated at 37 0
C for 30 min
& the absorbance of each solution was measured at 490 nm, using Elisa reader against the corresponding test & standard blanks, & the remaining DPPH was calculated. IC50 is the concentration of the sample required to scavenge 50% of DPPH free radicals.
RESULTS:
The experiments were repeated in triplicate and the average values were taken as shown in table 1, which shows the inhibitory activity of the essential oil against the selected strain of organisms.
Antibacterial activity of essential oil is respectable with that of standard drugs. The pure oil showed maximum activity against S. aureus with zone of inhibition 24mm and least activity against B. subtilis with zone of inhibition 20mm, where as DMSO does not show any zone of inhibition
By Nitric acid method the extract shows antioxidant activity with IC
50
value 121

8.25
 g/ml where as with DPPH method the IC
50
value was found to be 602.17

5.11
 g/ml.

Table 1: Antibacterial activity of essential oil of Lemon grass leaves:
Sr.
No.
1.
Organism
Tested
S. aureus
Diameter of Zone of Inhibition (in mm)
E. oil
26
Standard
30 *
2.
B. subtilis 20 28 *
3.
E. coli
4.
P. aeruginosa
25
22
34 **
32 **
*Penicillin (100 µl/ml), **Streptomycin (100 µl/ml).
DMSO does not show any zone of inhibition.
Table 2: Antioxidant activity of essential oil:
Essential oil Antioxidant activity (IC
50
)

Cymbopogon flexuousus
Nitric acid Method
121

8.25
 g/ml
DPPH Method
602.17

5.11
 g/ml.
CONCLUSION:
The essential oil of Lemon grass leaves exhibited a good activity against both gram positive and gram-negative organisms. The oil also exhibits antioxidant activity by Nitric oxide radical inhibiting activity and 2,2-diphenyl-1-pickryl hydrazyl (DPPH) method.
FUTURE PLAN:
To evaluate the essential oil of Lemon grass leaves for in-vivo antibacterial and antioxidant activity.
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