Indian Journal of Sciences ISSN: 2319–3824(Online) An Open Access, Online International Journal Available at http://www.cibtech.org/jps.htm 2016 Vol.5 (3) July-September, pp. 13-17/Powar and Gaikwad Research Article GC-MS ANALYSIS OF BIOACTIVE COMPOUNDS OF CORNICULATUM BARK *P. Powar1 and D. Gaikwad2 1Department of Botany, Rajarshi Chhatrapati Shahu College, Kolhapur, 416004, MS 2Department of Botany, Shivaji University, Kolhapur, 416004, MS India *Author for Correspondence

ABSTRACT India is a rich source of as it surrounded by Arabian Sea to Southwest, Bay of Bengal to Southeast and Indian Ocean to South. The methanolic extracts of outer bark of (Fam. ), was screened by GC-MS method. It was revealed that out of ten bioactive compounds, five are in higher concentration namely 7-Tridecanone, Glycerin, 2 Heptanone-3 methyl alpha-L-Galactopyranoside, methyl 6-deoxy- and 1,6-Anhydro-beta-D-glucopyranase (levoglucosan) while 2-Furan carboxaldehyde, 5 methyl, 4H-Pyran-4 –one, n-Hexadecanoic acid, Benzene, (2,2-dimethoxyethyl), Hexadecanoic acid methyl ester are found in lower concentrations. The antimicrobial properties of these secondary metabolites from other extracts have also been reported. The presence of these compounds in the methanol extracts could be used as antifungal, antibacterial and antioxidant agents. This mangrove can be used as wide range of bioresources that have rich source of photochemical.

Keywords: Aegiceras Corniculatum, Antimicrobial, Bark, GC-MS, Mangroves

INTRODUCTION Mangroves are having salt-tolerant capacity. They are found in both tropical and subtropical intertidal regions of the world. The regions where these plants dominated named as 'mangrove ecosystem'. Mangrove forest shows a tidal wetland tropical forest ecosystem with a very special association of flora and fauna that live in the intertidal zones. Mangroves are rich sources of variety of bioactive compounds. Kokpol et al., (1990) and Bandaranayake (2002) observed that Mangroves have been used in folklore medicines and extracts displays well-known activities against human, animal and plant pathogens. Secondary metabolites like alkaloids, phenolics, steroids and terpenoids have been characterized from mangroves which have toxicological, pharmacological and ecological importance. Indian coastline is about 5,700 km long among them Maharashtra measures 840 kilometers in Arabian Sea commonly known as Konkan. Konkan is widely distributed by six districts namely, Thane, Greater Mumbai, Navi Mumbai, Raigad, Ratnagiri and Sindhudurg. Ratnagiri is a in rich vegetation of mangrove Aegiceras corniculatum commonly known as black mangrove find in large scale in estuaries of Ratnagiri and Sindhudurg region (Jagtap et al., 1994; Bhosale, 2005) traditionally used for rheumatism, arthritis, inflammatory, diabetes and hepatoprotective actions (Roome et al., 2008). Therefore, present study was under takes to find out phytochemical compounds in the bark of stem of Mangrove Aegiceras corniculatum.

MATERIALS AND METHODS Collection of Sample The bark samples of mangroves of Aegiceras corniculatum from Family Primulaceae was collected from estuaries of Ratnagiri Districts in the month of December. Preparation of Powder and Extract Stem bark was shade dried, powdered and extracted with Methanol for 6-8 hours using Soxhlet apparatus. The extract was then filtered through muslin cloth, evaporated dried to get the viscous residue. The methanolic extracts of the plant was used for GC-MS analysis. 1μl of the methanolic bark extract of stem

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Indian Journal of Plant Sciences ISSN: 2319–3824(Online) An Open Access, Online International Journal Available at http://www.cibtech.org/jps.htm 2016 Vol.5 (3) July-September, pp. 13-17/Powar and Gaikwad Research Article was employed for GCMS analysis. The analysis of Mass spectra was done by retention time (RT) and library search of National Institute of Standards and Technology (NIST).

RESULTS AND DISCUSSION The mass spectra of Aegiceras corniculatum exhibits ten important peaks out of which four peaks were having larger percent area (Table 1 and Figure 1) with 7-Tridecanone, Glycerine, 2-Heptonone-3 methyl, and α-L-Galactopyranoside methyl 6-deoxy- metabolites showing percent area 37.26%, 11.82%, 10.09% and 9.68% respectively. There are six peaks shows lower percent area 1,6-Anhydro-beta-D-glucopyranase (levoglucosan), 2-Furan carboxaldehyde, 5 methyl, 4H-Pyran-4 –one, n-Hexadecanoic acid, Benzene, (2,2-dimethoxyethyl) and Hexadecanoic acid methyl ester with percent area 8.33, 6.98, 5.51, 4.92, 3.42 and 2.00 respectively. The various phytochemicals which contributes to the pharmaceutical activity of the plant Aegiceras corniculatum showed in Table 2, the phyto-constituents like 2-Furan carboxaldehyde, 5 methyl and 4H- Pyran-4 -one shows antimicrobial and anti-inflammatory potentials, n-Hexadecanoic acid and Hexadecanoic acid methyl ester are known for their Antioxidant, Hypocholesterolemic, Nematicide, Pesticide, Lubricant, Flavor. Glycerin was common to pharmaceutical and cosmetic industry as it used as base material. Mangrove plants have been used in folklore medicines and extracts from mangrove have verified activity against human, animal and plant pathogens. Mangroves are the rich sources of variety of bioactive compounds. Mangroves are growing in stress conditions so secondary metabolites like alkaloids, phenolics, steroids and terpenoids have been reported from mangrove and have toxicological, medicinal and biological importance (Bandaranayake, 2002 and Kokpol et al., 1990). Several reports prove that mangroves have antimicrobial properties associated with isothiocyanates (Dornberger et al., 1975 and Iwu et al., 1991), thiosulfinates (Tada et al., 1988) and glycosides (Murakami et al., 1993; Rucker et al., 1992). Polyacetylenes also have special attention due to organic polymer with repeating unit. Estevez- Braun et al., (1994) isolated C17 polyacetylene compounds form Bapleurum salicifolium a plant native to the canory Islands. The compound 8S-Heptadeca-2(z), 9(z) diene-4, 6-diyne-1, 8-diol, was inhibitory to Gram negative bacteria, Staphyllococcus aureas and Bacillus subtilis but not to gram positive bacteria and Yeasts. The mature of A. marina (Abeysinghe and Wanigatunge, 2006) and Aegiceras corniculatum (Poompozhil and Kumarasamy, 2014) contain alkaloids, steroids, triterpenoids and flavonoids. The bark of A. corniculatum is also rich in Polyphenols, flavonoids and tannins (Powar and Gaikwad, 2014). Phytochemical investigation of stylosa Griff. (Rhizophoraceae) showed one new acetylated flavanol, 3, 7, 0-diacetyl (-) epicatechin (Anjaneyulu et al., 2002). The bark of A. corniculatum have very good antifungal and antibacterial activities against various strains of fungi, Alternaria alternata and Fusarium moniliforme (Powar et al., 2011) and bacteria, Bacillus megaterium, Staphyllococcus aureus, Erwinia carotovora and Xanthomonas citri (Powar et al., 2009). This activity of A. corniculatum might be due to presence of antimicrobial compounds like 2-Furan carboxaldehyde, 5 methyl and 4H-Pyran-4 –one. The main chemical compounds isolated from the bark of Bauhinia variegate are quercitroside, isoquercitroside, rutoside, myrcitol glycoside and kamferolglycoside (Gupta et al., 1980). Gupta et al., (1978) isolated 5, 7-dihydroxyflavanone 4’-02-1 rhamnopyranosyl-β-D-glucopyranoside (I) from the stem. The stem is rich source of β-sitosterol, lupeol, Kempferol-3-glucoside and 5, 7-dimethyl ether 4’ rhamnoglucoside. The stem barks of Bauhinia variegate yields four substances viz. Hentriacontane, Octacosanol, β-sitosterol and Sigmasterol (Anandaprakash, 1978). Glycerin in combinations with other phytochemicals dimethicone, petrolatum, antioxidants, fatty acids, lecithin are useful in healing of skin and cures dermatitis (Lorancini et al., 2014). Phytochemical analysis of the methanolic bark extract of mangrove A. corniculatum revealed that presence of variety of phytoconstituents Glycerine, 2-Furan carboxaldehyde, 5 methyl, 4H-Pyran-4 –one, n-Hexadecanoic acid and Hexadecanoic acid methyl ester. These active chemical constituents have good

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Indian Journal of Plant Sciences ISSN: 2319–3824(Online) An Open Access, Online International Journal Available at http://www.cibtech.org/jps.htm 2016 Vol.5 (3) July-September, pp. 13-17/Powar and Gaikwad Research Article antimicrobial, anti-inflammatory, antiallergic and antioxidant properties. These secondary metabolites from other plants parts have also been reported earlier. It could be concluded that the presence of these compounds in the methanolic bark extract of Aegiceras corniculatum might be contents the antifungal, antioxidant, anti-inflammatory pesticide and nematocidal bioactive phytochemicals. Further study is essential to isolate individual phytochemical constituents and subjecting it to the biological activity will give productive results. This plant may be used as one of the good sources of pharmaceutical industry. This mangrove can be used as wide range of bioresources that have rich source of phytochemicals. Thus, the barks of this plant can be successfully used for the synthesis of anti-inflammatory, antimicrobial, analgesic drugs with further careful investigations and it will also enhances knowledge about use of traditional medicine.

Table 1: Relative Percentage Composition of Bark Extract of Mangrove Aegiceras Corniculatum Sr. Retention Constituents Molecular Molecular Concentration No. Time Formula Formula % (Min.) 1 11.227 7-Tridecanone C13H26O 198 37.26 2 9.739 Glycerin C3H8O3 92 11.82 3 12.400 2 Heptanone-3 methyl C8H16O 128 10.09 4 19.698 alpha-L-Galactopyranoside, C7H14O5 178 9.68 methyl 6-deoxy- 5 23.589 1,6-Anhydro-beta-D- C6H10O5 162 8.33 glucopyranase (levoglucosan) 6 9.390 2-Furan carboxaldehyde, 5 C6H6O2 110 6.98 methyl 7 14.038 4H-Pyran-4 -one C6H8O4 144 5.51 8 31.283 n-Hexadecanoic acid C16H32O2 256 4.92 9 15.916 Benzene,(2,2-dimethoxyethyl ) C10H14O2 166 3.42 10 30.827 Hexadecanoic acid methyl ester C17H32O2 270 2.00

Table 2: Activity of Phyto-Components Identified in the Methanolic Extracts of the Bark of Aegiceras Corniculatum by GC-MS Sr. Name of the Compound Nature Activity No. Compound 1. Glycerin Polyol coumpound Skin healing, cures dermatitis (Lorancini et al., 2014) 2 2-Furan Aldehyde **Antimicrobial, Preservative carboxaldehyde, 5 compound (Ravikumar et al., 2012)

methyl 3 4H-Pyran-4 -one Flavonoid fraction **Antimicrobial, Anti inflammatory 4 n-Hexadecanoic acid Palmitic acid Antioxidant, Hypocholesterolemic, Nematicide, Pesticide, Lubricant, Flavor, Hemolytic 5- Alpha reductase inhibitor (Rajeswari et al., 2013) 5 Hexadecanoic acid Palmitic acid Antioxidant, Hypocholesterolemic methyl ester Nematicide, Pesticide, Lubricant, Flavor, Hemolytic 5-Alpha reductase inhibitor (Rajeswari et al., 2013) **Source: Dr.Duke's phytochemical and ethnobotanical databases [Online database].

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Indian Journal of Plant Sciences ISSN: 2319–3824(Online) An Open Access, Online International Journal Available at http://www.cibtech.org/jps.htm 2016 Vol.5 (3) July-September, pp. 13-17/Powar and Gaikwad Research Article

Figure 1: GC Ms Spectra of Methanolic Extract of Aegiceras Corniculatum

ACKNOWLEDGEMENT One of the authors is very much thanking to head department of Botany and CFC department Shivaji University, Kolhapur, MH –India for providing all needful things for this piece of research work.

REFERENCES Abeysinghe PD and Wanigatunge RP (2006). Evaluation of antibacterial activity of different mangrove plant extracts. Ruhuna Journal of Science 1 104-112. Anandaprakash RL (1978). Natural constituents of Bauhinia variegata. Journal of Research into Indian Medicine, Yoga and Homeopathy 13 96-97. Anjaneyulu ASR, Anjaneyulu V and Rao VL (2002). New Beyerane and Isopimarane Diterpenoids from Rhizophora mucronata. Journal of Asian Natural Products Research 4 53-61. Bandaranayake WM (2002). Bioactivities, bioactive compounds and chemical constituents of mangrove plants. Wetland Ecology and Management 10 421-452. Bhosale LJ (2005). Field Guide to Mangroves of Maharashtra, published by (Shivaji University, Kolhapur (MS), India). Dornberger K, Bockel V, Heyer J, Schonfeld C, Tonew M and Tonew E (1975). Studies on the isothiocyanateserysolin and sulforaphan and from Cardariadraba. Pharmaizie 30 792-796. Duke J (1994). Dr. Duke's Phytochemical and Ethnobotanical Databases (Online). USDA-ARS-NGRL- Beltsville Agricultural research institute, Maryland. Available: http//www.ars-grin.gov/cgi- bin/duke/ethanobot.pl (Accessed 25 July 2016). Estevez-Braun A, Estevez-Reyes R, Moujir LM, Ravelo AG and Gonzalez AG (1994). Antibiotic activity and absolute configuration of 8S-heptadeca-2(Z), 9(Z)-diene-4, 6-diyne-1, 8-diol from Bupleurum salicifolium. Journal Natural Product 57 1178-1182. Gupta AK, Vidyapati TJ and Chauhan JS (1980). Chemical examination of Bauhinia variegate. Phytochemistry 64 879-882. Gupta YK, Gupta MN and Roy AN (1978). Reaction of sunflower varieties to Alternaria alternate. Indian Phytopathology 31 507-508. Iwu MM, Unaeze N C, Okunji CO, Corely DG, Sanson DR and Tempesta MS (1991). Antimicrobial aromatic isothiocyanates from the essential oil of Hippocratea welwitschii roots. International Journal of Pharmacognosy 29 154-158. Jagtap TG, Untawale AG and Inamdar SN (1994). Study of mangrove environment of Maharashtra coast using remote sensing data. Indian Journal of Marine Sciences 23 90-93.

Centre for Info Bio Technology (CIBTech) 16

Indian Journal of Plant Sciences ISSN: 2319–3824(Online) An Open Access, Online International Journal Available at http://www.cibtech.org/jps.htm 2016 Vol.5 (3) July-September, pp. 13-17/Powar and Gaikwad Research Article Kokpol U, Miles DH, Payne AM and Chittawong V (1990). Chemical constituents and bioactive compounds from mangrove plants. In: Att-ur- Rahman (edition), Studies in Natural Products Chemistry, (Elsevier Science Publishers B. V, Amsterdam, Netherland) 7 175-195. Lorencini M, Brohem C, Dieamant G, Zanchin N and Maibach H (2014). Active ingradients against human epidermal ageing. Research Review 15 100-115. Murakami A, Ohigashi H, Tanaka S, Tatematsu A and Koshimizu K (1993). Bitter cyanoglucosides form Lophiraalata. Phytochemistry 32 1461-1466. Poompozhil S and Kumarasamy D (2014). Studies on phytochemical constituents of some selected mangroves. Journal of Academia and Industrial Research 2(10) 590-592. Powar PS and Gaikwad DK (2014). Quantification of phytochemicals from some mangrove bark Indian Journal of Advances in Plant Research 1(1) 5-9. Powar PS, Gaikwad DK and Chavan PD (2009). Antibacterial activity of mangrove bark. Bioinfolet 6(4) 365-367. Powar PS, Gaikwad DK and Chavan SR (2011). Antifungal activity of mangroves bark. International Journal of Pharma and Bio Sciences 2(4) 484- 488. Rajeswari G, Murugan M and Mohan VR (2013). GC-MS analysis of bioactive components of Hugonia mystax L. bark (Linaceae). Journal of Pharmceutical Biomedical Sciences 29 818-824. Ravikumar VR, Gopal V and Sudha T (2012). Analysis of Phytochemical Constituents of Stem Bark Extracts of Zanthoxylum Tetraspermum Wight & Arn. Research Journal of Pharmaceutical, Biological and Chemical Sciences 3(4) 391-402. Roome T, Dar A, Ali S, Naqvi S and Chuodhary MI (2008). A study on antioxidant, free radical scavenging, anti-inflammatory and hepatoprotective actions of Aegiceras corniculatum (stem) extracts. Journal of Ethanopharmacology 118(3) 514-521. Rucker G, Keherbanum S, Sakulas H, Lawong B and Goeltenboth F (1992). Acetylenic glucosides from Microglos sa pyrifolia. Planta Medica 58 266-269. Tada M, Hiroe Y, Kiyohara S and Suzuki S (1988). Nematicidal and antimicrobial constituents from Allium grayi Regel and Allium fistulosum L.var. caespitosum. Agricultural and Biological Chemistry 52 2383-2385.

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