IDENTIFICATION OF BIOACTIVE COMPOUNDS AND …

1 , (2)2012:263-266 ISSN2229 6441263 IDENTIFICATION OF BIOACTIVE COMPOUNDS AND ANTIMICROBIALACTIVITY OF MARINE CLAMANADARAGRANOSA(LINN.)Ramasamy, , and Research department of zoology and Biotechnology, Sri Pushpam college(Autonomous)Poondi 613 503 Thanjavaur district,Tamilnadu, South inventory of the BIOACTIVE COMPOUNDS produced byAnadara granosaduring full growth has been establishedby whole tissue using organic solvents system and subjecting in to gas chromatography mass spectrum (GC-MS) twelvebioactive COMPOUNDS of which ester, furan, acid and ketone were identified. The most representative BIOACTIVE COMPOUNDS ofAnadara granosawere employed in the antibacterial activity against pathogenic bacteria and the studypertaining to theidentification of active principles and the antimicrobial WORDS:Anadara granosa;bioactivecompounds; Gas chromatography and mass spectrum; granosa,belongs to the family Arcidae.

2 It ispopularly known as blood clam due to the presence ofhaemoglobin which gives red colour to the meat(Narashiman,1988). It is widely distributed in theindopacific, from Eash Africa to Polynesia, North Japan andSouth to Northern and Eastern Australia (Poutiers, 1988) andalso widely distributed along the Indian coast and form afishery of considerable magnitude of Bay of Bengal theocean was a vast potential for a huge number of novelchemicals that may useful for finding drugs with greaterefficacy and specificity for the treatment of many humandiseases (Bergman and Feeney, 1951, Faulkner, 2001).

3 Themarine organism have to with stand extreme variation inpressure, salinity, temperature etc., and these environmentalvaluables have facilitated the organismto produced variedchemicals of unique features. So far more than 10,000compounds have been isolated from marine organisms(Prokschet al., 2002) with hundred of near COMPOUNDS stillbeing discovered every year (Proskchet al., 2002).Extensive Biochemical studies have been carried out inwhole body tissue ofAnandra there are noreports on the Biochemical aspects related withantimicrobial activity against various AND METHODSM arine clamAnadara granosawere collectedfromMuthupet estuary (Latitude 10 20 N, Longitude 79 32 E),South East Coast of India and identified using the standardreference (Sathiyamoorthy, 1952) and also confirmed withzoological survey of India.

4 50 gm sample were dissected outand cleaned in distilled water. Extracts are homogenizedwith organic solvent a mixture of (1:1 v/v) pentane: diethylether. The upper organic phase were separated, dried overanhydrous sodium sulphate, and concentrated at 42 C to 1ml and at 20 C for antibacterial assay and GC-MS strain and culture conditionAntibacterial activity of marine clam was determined againstfour bacterial strain ,Streptococcus faecalis,Escherichia coli, Pseudomonasaeruginosathese pathogens were isolated from patients ofRaja Mirasudar Hospital, Thanjavur, South of bacterial cultureNutrient broth medium was prepared and sterilized in anautoclave at 151lb and the presence of four bacterial specieswere inoculated in the nutrient broth and incubated at 28 2 C for 24 hrs.

5 On this 24 hrs old bacterial broth cultureswere inoculated by using a sterile colony chromatography Mass spectrum (GC-MS)Volatile component were identified by GC-MS using GCClarus 500 PerkinElmer (Turbo mass gold Perkin Elmer)equipped with column Elite 1 (30m mm 1) nmdscarrier gas (Hon rate, ml/min) Helium was used andinfection volumes were 1m. The column temperature wasmaintained initially at 400 C for min followed byincrease to 600 C at a rate of 50 C/min from 60 to 120 C ata rate of 60 C/min and from 120 to 230 C. The electronimpact energy was 70 ev and the ion source temperature asset at 2300 C.

6 Electron impact (EI) Mass spectra wererecorded in the 45-450 a Mu range at 1s activity was tested against four bacteria amongthe bacterial antibacterialactivity of clam extract issummarized inTable 1 the level of activity which was measured byinhibition zone, variedbetween 10 to 16 mm. The zone ofinhibition showed highest activity againstPseudomonasaeruginosa( ). antimicrobial activity of marine clamAnadaragranosa264 TABLE of inhibition (mm) of clams crude extract against 4 pathogenic of inhibitionClams extract (mm)Standard (mm) aeruginosa16170246810121416182022 StaphylococcusaureusStreptococcusfaecali sE.

7 ColiPseudomonasaeruginosaZone of inhi bi tion (mm)PathogenClams of clams extract against four Gas Chromatography and Mass Spectra forAnadara granosaIdentification of BIOACTIVE , (2)2012:263-266 ISSN2229 6441265A typical chromatogram from clam extract obtained in theGC/MS system underprogrammed temperaturecondition isshown in BIOACTIVE COMPOUNDS were detected. Theyare Furancompound, Palmitic acid, Fatty acid esters, Stearic acid,Ketone, Phenol compound and Plasticizer of clam GC-MS analysis showed thatAnadragranosahas a complex fatty acid ester compound . Fromwhich 4, 7, 10, 13, 19.

8 Doca sahexaenoic acid hasantidiabetic, antiasthma, anticancer and anti heart diseasesproperties Retention time phenol, 2-4 bis (1-phenylethyl) has analgesic Anasthedic, Antioxidantantiseptic, antibacterial, antitumor and cancer preventiveproperties (Table2).TABLE composition in clamAnadara of the compoundMolecularformulaMWPeakarea %CompoundnatureActivity** ,3-dihydrobenzo[b] 5-(2-phenylpropionyl) acidAntioxidant acid, acid acid, methyl acid esterNo activity acidNo activity acid, ethyl acid esterNo activity , (1,4-dimethyl-7-(1-methylethyl)-2-azulen yl) activity , [2-(1-methylethyl)Phenyl] activity ,7,10,13,16,19-Docosahexaenoic acid,methyl ester, (all-z) acid esterAntidiabeticAnti asthmaAnticancerAnti heart , 2,4-bis (1-phenylethyl)

9 ,Anesthetic,Antioxidant,Antiseptic,Antib acterial,AntiviralDiureticCancer preventive,Fungicide,RodenticideEmetic, ,2-Benzenedicaryboxylic acid, foulingDISCUSSIONIn the present study, GC-MS analysis and antibacterialactivityAnadara granosashowed 100 per cent success inthe IDENTIFICATION of BIOACTIVE COMPOUNDS responsible forantimicrobial and anticancer activities. Whole tissue extractofAnadara granosashowed broad spectrum of antibacterialactivity (Table 1). The secretion of hypobranchial gland ofmurcid Rapana rapiformis, was found to inhibit eightbacteria (Muruganet al., 1991).

10 Sincerely the hypobranchialgland ofC. ramosusinhibited a broad spectrum of activityagainst ten bacterial strain (Kagoo and Ayyakkannu, 1992).Many classes of BIOACTIVE COMPOUNDS exhibiting antitumor,antileukemia, antibacterial and antiviral activities have beenreported sterol composition of clamAnadara granosais shownin table 2 among 6 sterols identified in this study 4, 7, 10,13, 19 Docosahexaenoic acid is the major sterolcomponent, which were similar to those observed in thebivalveMacomabalthicafrom the Baltic sea (Pazoset al.,1992). These sterol composition ofAnadara granosawasvery similar to that maximusand the pacific oysterCrassostrea giga(9,10) (Idleret al.)