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Int J Pharm Bio Sci 2013 July; 4(3): (B) 380 - 384

This articl can ! "o#nloa"" $rom ###%i&'!s%ntB - 380

sarch rticl *icro!iolo+y

Intrnational Journal o$ Pharma an" Bio Scincs ISS,

0./-2

PI*I, PT56*I6 S6,I,7 , ,TIB6TI

ST9IS 5 T S 5 6IPT B ()

VENKANNA LUNAVATH AND ESTARI MAMIDALA*

Metabolic Disorders and Infectious Diseases Research Lab,

Department of Zoology, Kakatiya University, arangal!"#$##%, &'()*, India

ABSTRACT

Globally, medicinal plants have been used for treatment of several ailments. Thepresent study was performed to evaluate anti microbial activity and preliminaryphytochemical screening of Eclipta alba(L) leaf extracts. The Eclipta alba (L.)belongsfamily Asteraceae. The Eclipta alba were shade dried, powered and was extractedusing solvents ethanol and other solvents. The antimicrobial activity test performed bythe disc diffusion method. !reliminary phytochemical analysis of the plant extractsfractions of "arbon tetrachloride fraction ("T#), "hloroform fraction( "##) andA$ueous fraction(A%#) showed the presence of al&aloaids, tannin, flavonoidsphenolic group and flavonoids. The "T# fraction of E. alba showed high activityagainst E. coliand E. faecalisbacteria.

KEYWORDS: Antimicrobial activity, Eclipta alba, disc diffusion, MIC.

*Corresponding author

hESTARI MAMIDALA

*ta!olic isor"rs an" In$ctious isass sarch a!?#, "T#, "## and A%#) were evaporatedto dryness by using a rotary evaporator at lowtemperature (76").

(ii) Preliminary phytochemicalinvestigatons!hytochemical screening of plant extractswere carried out $ualitatively for the presenceal&aloaids, tannin, flavonoids phenolicgroup and flavonoids were screenedaccording to the common phytochemicalmethods described by 'o&ate (-662) and'o&ate et al(-66@).

(iii) Bacterial species

acterial species selected for the study werethe four pathogens, namely, two Gram5positive Staphylococcus aureus andEnterococcus faecalisand two Gram5negativePsuedomonas aeuruginosa and Escherichiacoli. All the cultures were maintained onueller5>ilton agar at 2". The cells wereinoculated and incubated at 7*" in broth for- hours prior to the screening procedure.

(iv) Minimal inhibitory concentration (MIC)The serial microplate dilution method

developed by 3loff (-66) was used todetermine the minimum inhibitoryconcentration (0") for plant extracts usingtetra1olium violet reduction as an indicator ofgrowth. :esidues of the different extracts werere5dissolved in methanol to a concentration of- mgBml. for each of the four bacteria used,-Cl of each plant extract tested were two5fold serially diluted with - Cl sterile distilledwater in a sterile 6D5well microtitre plates. A

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similar two5 fold serial dilution of gentamicine(.-mgBml) was used as a positive controlagainst each bacterium. =ne hundredmicrolitres of each bacterial culture wereadded to each well. The plates were coveredand incubated overnight at 7* ". To indicatebacterial growth 2 Cl of . mgBml p5

iodonitrotetra1olium violet (0ET) were added toeach well and the plates incubated at 7* " for7 minutes. acterial growth in the wells wasindicated by a red colour, where as clear wellsindicated inhibition of the bacterial growth bythe plant extracts.

RESULTS AND DISCUSSION

(v) Percentage yieldThe yield of the methanol crude extract ofEclipta alba was @gr (-D.2F) . Thepercentage yield of these fractions of themethanolic extract of Eclipta alba were

showed in the table5-. The "T# fractionsobtained highest yield (.F) when comparedto other fractions. .F yield obtained in >?#fraction which is lowest.

Table-1Percentage of yield

S.No Fractions Yield !"

- >?# .

"T# .

7 "## -.@

2 A%# -.

(vi) Phytochemical screening in E. lbaThe phytochemical analysis of Eclipta alba showed the presence of different groups ofsecondary metabolites vi1, al&aloaids, tannin, flavonoids which are of medicinal importanceof the test extracts, a$uous fraction showed positive results for most of the test compounds.The phenolic group and flavonoids were rich in "T#, "## and A%# fractions whencompared to other metabolites (Table 5). Al&aloids are rich in all fractions of E. alba.

Table-#Phytochemical screening for presence of different phytoconstituents

in Eclipta alba leaves e!tract fractions

Sl. No. $%ytoc%emicals test &'F CTF CFF A(F

I Test or Al)aloids

a. ayers Test * * * *

b. 9agners Test * * * *

c. >agers Test ** ** ** ***

+ ;ragendorffs Test * * * *

II Test for Carbo%ydrates

A olishs Test * * ** ***

#ehlings Test - - ** ***

" arfoeds Test - - ** ***

; enedicts Test - - * **

III Test for lycosides

a. orntragers Test * * * **

Legals Test * * ** ***

I Test for Saonin

A #oam Test * *** *** -

Test for $roteins and Amino acidsA illons Test - ** * -

iurets Test - ** ** -

" Einhydrin Test - ** * -

I Test for $%ytosteroids

A Libermann H urchards Test - - *** **

II Test for fi/ed oils and fats

A 4pot Test * ** - **

4aponification Test - * - ***

III Tests for $%enolic Comounds and Flavanoides

A #erric chloride Test - * * -

Gelatin Test - * - -

" Lead acetate Test - * - **

; Al&aline :gt. Test - ** ** *

3 agnesium Test - ** * *

""" Prominently Present# "" Moderately Present# " $lightly Present# % bsent

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(vi) ntibacterial activityThe 0" values and total activity of the four fractions of methanol crude extract of Eclipta albaplant against all the tested bacteria are presented in Table 7. E. alba A%# fractions also hadnoteworthy 0" values of . mgBml and .D7 mgBml against S. aereus and E. faecalisrespectively

Table 0

MIC values m2ml" of t%e four lant e/tracts fractions of t%e study3 P. emblica# E. alba# &. cordifoliaand C. occidentalis aainst four bacteria' Escherichia coli 4c", Enterococcus faecalis 4f",Pseudomonas aeuruginosa $a" and$taphylococus aureus Sa".entamicin5as used as a ositivecontrol.

$lant secies Ec Ef Pa $a

E. alba>?#"T#"##A%#

[email protected]@[email protected]

[email protected]

.D7.7--.@-.@

[email protected]

Gentamicin (CgBml) . -.D . .7

+n%he!ane fraction# C&+Carbon tetra chloride fraction# C+Chloroform fraction and ,+-ueous fraction.

0n this study the highest total activity was obtained on Eclipta alba (Table52) from >?#fraction across Enterococcus faecalis (3f), Pseudomonas aeuruginosa (pa) andStaphylococus aureus (4a) bacteria. "T# fraction of E. alba showed high total activityagainst E. coli(-2D mlBmg) and E. faecalis(-2D mlBg) bacteria.

Table-6Total activity ml2" of four lant e/tracts fractions of t%is study3 P. emblica# E. alba# &. cordifoliaand C occidentalis aainst four bacteria' Escherichia coli 4c", Enterococcus faecalis 4f",Pseudomonas aeuruginosa $a" and$taphylococus aureus Sa".

$lant secies Ec Ef Pa $a

E. alba>?#"T#

"##A%#

2-2D

---@

---2D

-@

--*

--

--72

---@

+n%he!ane fraction# C&+Carbon tetra chloride fraction# C+Chloroform fraction and ,+-ueous fraction.

DISCUSSION

The primary and secondary metabolites wereanaly1ed in methanolic extracts. 0n an earlierstudy the extracts of the leaves of E. albatested positive for al&aloids, phenolics,saponins and tannins, but no anthra$uinonesand flavonoids were detected ("aldentey et

al.,2). 0n Gurat and !unab, E. alba is usedexternally for ulcers and as an antiseptic forwounds in cattle and is reported to treat manymicrobial infections in rural areas (9arrier,-662). The results from the current studiesrevealed that the wedelolactone may be themain constituent responsible for antimicrobialactivity. There are various reports that crudeextract from E. alba showed antibacterial,antifungal and anti viral activity ('osuge etal.,

-6@+ 9iart et al., 2+ 'arthi&umar et al.,*). 0n this study the "T# fraction of E.alba showed high activity against E. coliand E. faecalisbacteria.

CONCLUSION

;ifferent extracts from the aerial parts ofEclipta alba showed antimicrobial activityagainst four microbial species. ost of theantimicrobial compounds of Eclipta alba aresoluble in butanol and ethyl acetate. Theantimicrobial compounds present in Ecliptaalba may serve as an affordable and newsource for the treatment of infectious diseases.

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