Â In the present work, we used two seaweeds Ulva lactuca and Gracilaria corticata, which were collected from Hare Island in the Gulf of Mannar ofTuticorin coast. Ethanol was taken as the solvent for extraction. The crude extract was purified using column chromatography. Antibacterial activityof crude and column purified fractions were tested against Klebsiella, Aeromonas, Staphylococcus, Escherichia and Pseudomonas using well-diffusionmethod. Maximum zone of inhibition (9 mm) was found in the crude extract of G. corticata against Pseudomonas sp. Minimum zone of inhibition (4mm) was found in U. lactuca fraction1 against Escherichia coli. Highest antibacterial activity was obtained in red seaweed, whereas, green seaweedshowed less antibacterial activity. From this study, we can conclude that red seaweeds have more active than green seaweeds. The seaweed powderwas analyzed in Fourier transform infrared spectrometer. It was observed that both seaweed contained phenol and alcohol compounds, which wereresponsible for the antibacterial activity.Keywords: Antibacterial activity, Fourier transform infrared, Seaweeds, Ulva lactuca, Gracilaria corticata.Â

A, Mohaideen

RADHIKA D

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Radhika D

Mohaideen A

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FOURIER TRANSFORM INFRARED ANALYSIS OF ULVA LACTUCA AND GRACILARIA CORTICATA AND THEIR EFFECT ON ANTIBACTERIAL ACTIVITY

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Vol 8, Issue 2, 2015 ISSN - 0974-2441FOURIER TRANSFORM INFRARED ANALYSIS OF ULVA LACTUCA AND GRACILARIA CORTICATA AND THEIR EFFECT ON ANTIBACTERIAL ACTIVITYRADHIKA D*, MOHAIDEEN ADepartment of Zoology PG and Research, V.O.C. College, Tuticorin, Tamil Nadu, India. Email: drradhi24@gmail.comReceived: 20 November 2014, Revised and Accepted: 02 January 2015ABSTRACTIn the present work, we used two seaweeds Ulva lactuca and Gracilaria corticata, which were collected from Hare Island in the Gulf of Mannar of Tuticorin coast. Ethanol was taken as the solvent for extraction. The crude extract was purified using column chromatography. Antibacterial activity of crude and column purified fractions were tested against Klebsiella, Aeromonas, Staphylococcus, Escherichia and Pseudomonas using well-diffusion method. Maximum zone of inhibition (9 mm) was found in the crude extract of G. corticata against Pseudomonas sp. Minimum zone of inhibition (4 mm) was found in U. lactuca fraction1 against Escherichia coli. Highest antibacterial activity was obtained in red seaweed, whereas, green seaweed showed less antibacterial activity. From this study, we can conclude that red seaweeds have more active than green seaweeds. The seaweed powder was analyzed in Fourier transform infrared spectrometer. It was observed that both seaweed contained phenol and alcohol compounds, which were responsible for the antibacterial activity.Keywords: Antibacterial activity, Fourier transform infrared, Seaweeds, Ulva lactuca, Gracilaria corticata.INTRODUCTIONSeaweeds are multicellular macroalgae used as potential renewable resource in the field of medical and commercial environment, which are also used as food, feed and fertilizer in many parts of the world. Biostimulant properties of seaweeds are explored for use in the development of novel antibiotics. Many metabolites isolated from marine algae have bioactive efforts [1]. Bioactive natural products are widely distributed in the plant kingdom and extract from different plants, as well as red, green and brown macro, and micro algae can be used as natural products [2]. Marine algae represent an inexhaustible reservoir of raw materials used in pharmaceutical, medicine, food industries and cosmetics [3]. In recent years, research on the chemistry of seaweeds has experienced a tremendous increase due to the need for compounds possessing bioactivities of possible pharmaceutical applications or other potential economic properties [4]. Marine algae serve as an important source of bioactive natural substances [5]. Special attention has been paid to antibacterial activities related to marine algae against several pathogens [6]. The extracts and active constituents of various marine algae have been shown to have antibacterial activity against Gram-positive and Gram-negative bacteria [7]. The antimicrobial compounds derived from the marine algae consist of a diverse group of chemical compounds [8]. In the present study, antibacterial activities of Ulva lactuca was investigated against some fish pathogenic bacteria.METHODSCollection of seaweedLive samples of the seaweed U. lactuca and Gracilaria corticata was collected by handpicking during low tide from Hare Island in the Gulf of Mannar of Tuticorin coast (08°46’ 2.15”N latitude; 78°11’ 16.05”E longitude). Shade-dried seaweeds were ground to a fine powder using an electric mixture grinder. The powdered samples were then stored in refrigerator for further use.Preparation of extracts by soxhlet extraction methodThe powdered samples were extracted by using soxhlet apparatus. Ethanol was taken as the solvent for extraction. A total of 25 g of the sample and 250 ml of the solvent were taken for extraction the apparatus was run for 4 hrs and syrupy extracts were collected. The extracts obtained were concentrated by evaporation. Then, the extract was stored in cold storage for further use.Column chromatographyColumn chromatography is one of the most useful methods for the separation and purification of crude extract. The ethanolic crude extracts were applied in a silica gel column (230-400 mesh), packed with chloroform and eluted with a mixture of chloroform and methanol in the following proportions 9:1, 8:2 and 7:3. After purification, the fractions were stored at a temperature of −20°C until the determination of antibacterial activity.Well diffusion methodThe antibacterial activity of fraction and crude extracts of U. lactuca and G. corticata against five species of bacteria namely Klebsiella pneumonia, Staphylococcus aureus, Aeromonas hydrophilla, Escherichia coli and Pseudomonas sp. were determined by well diffusion method. The circular well of 3 mm diameter and 25 µl holding capacity was prepared using well cutter. Streak plate method was performed to seed pathogenic bacterial culture on the agar plates. Using the loop which had been flamed, cooled and dipped in the inoculums, continuous horizontal streaks were made in the solid agar plates. Different fractions of the seaweed extracts were then added to the wells. The clear labels of sample were marked on the plate. The plates were then incubated at 37°C for 24 hrs.Fourier transform infrared (FT-IR) analysisInfrared reflectance vibrational spectra were carried out on powdered samples using a spectrometer with instrument resolution of about (1/cm), in the wave number region (4000-400/cm) at room temperature [9].RESULTWhen the fractions of U. lactuca were tested against different pathogens like K. pneumonia, A. hydrophilla, S. aureus, E. coli and Pseudomonas sp. highest zone of inhibition was found in A. hydrophilla in Fig. 2 fraction. Similarly, when the crude extract of U. lactuca was tested against the same bacterial pathogen highest zone of inhibition was found in E. coli. The fractions of G. corticata were tested against different pathogens like K. pneumonia, A. hydrophilla, S. aureus, E. coli and Pseudomonas sp. Research ArticleAsian J Pharm Clin Res, Vol 8, Issue 2, 2015, 209-212 Radhika and Mohaideen 210highest zone of inhibition was found in K. pneumonia in F2 fraction. Similarly, when the crude extract of G. corticata was tested against the same bacterial pathogen highest zone of inhibition was found in Pseudomonas sp.FT-IR analysisThe FT-IR spectrum was used to identify the functional group of the active components based on the peak value in the region of infrared radiation. The crude powder of U. lactuca was passed into the FT-IR and the functional groups of the components were separated based on its peak ratio. The results of FT-IR analysis showed different peaks at 620.15 and 848.99 functional group is alkyl halides, 928.18 functional group is carboxylic acids, 1086.14 and 1144.12 functional group are aliphatic amines, 1384.42 functional group is nitro methane, 1428.88 functional group is aromatics, 1634.33 functional group is amides, 2295.04 functional group is nitriles and 3406.60 functional group are alcohols, phenols were present in U. lactuca sample (Table 2). Similarly, the crude powder of G. corticata was passed into the FT-IR and the functional groups of the components were separated based on its peak ratio. The results of FT-IR analysis showed different peak value they are 3321.46 functional groups are alcohols, phenols, 2925.49 functional group is alkanes, 2084.83 functional group are allenes, ketenes, is ocyanates, isothiocyanates, 1647.84 functional group isamides, 1471.30 functional group isaromatics, 1116.62 functional group isaliphatic amines, 874.41 functional group are primary and secondary amines, 750.98, 712.56, 657.10 and 617.26 functional group is alkylhalides were present in G. corticata sample (Table 4).DISCUSSIONSeaweed extracts showed various bio potential activities such as antibacterial [10]. Antibacterial activity of red, brown and green algae against both Gram-positive and Gram-negative bacteria has been established by several scientists [11]. Marine environment contains a source of functional materials, including polyunsaturated fatty acids, polysaccharides, essential minerals, vitamins, antioxidants, enzymes and bioactive peptides [12]. FT-IR is a valuable tool for measuring many chemical constituents in plants and seaweeds and it is used to reveal some qualitative aspects regarding the organic compounds [13]. In the present study, FT-IR is used to identify the functional group in the seaweeds. Alcohols are commonly found to have antimicrobial properties against both Gram-positive and Gram-negative bacteria [14]. Phenolic compounds exhibited good antimicrobial activities [15,16]. In the present study both seaweed contained phenol and alcohol compounds, which were responsible for the antibacterial activity. In the present study, U. lactuca is compared to G. corticata the G. corticata is more active.Fig. 1: Fourier transform infrared spectrum of Ulva lactucaTable 2: FT-IR absorption frequencies (cm−1), intensity estimation and functional group of seaweeds U. lactucaIR frequency (cm−1)Bond Functional groupsU. lactuca IR frequency (cm−1)3500-3200 O–H stretch, H–bondedAlcohols, phenols3406.602300-2200 C≡N stretch Nitriles 2295.041670-1600 C=O stretch Amides 1634.331500-1400 C–C stretch Aromatics 1428.881400-1300 N=O bend Nitromethane 1384.421250-1020 C–N stretch Aliphatic amines 1144.121250-1020 C–N stretch Aliphatic amines 1086.14950-910 O–H bend Carboxylic acids 928.18850-550 C–Cl stretch Alkyl halides 848.99690-515 C–Br stretch Alkyl halides 620.15U. lactuca: Ulva lactuca, FT-IR: Fourier transform infraredTable 1: The zone of inhibition (mm) of Ulva lactuca against different bacterial pathogensFractions K. pneumonia A. hydrophilla S. aureus E. coli Pseudomonas sp.F1 5±0.05 5±0.81 4±0.07 4±0 5±0.81F2 4±2.45 6±4.09 5±0.06 4±0.70 5±0.81F3 6±0.85 4±0.84 4±0.88 6±1.63 6±1.63Crude 6±2.44 6±1.63 5±4.08 7±4.89 6±2.44K. pneumonia: Klebsiella pneumonia, A. hydrophilla: Aeromonas hydrophilla, S. aureus: Staphylococcus aureus, E. coli: Escherichia coli, U. lactuca: Ulva lactucaAsian J Pharm Clin Res, Vol 8, Issue 2, 2015, 209-212 Radhika and Mohaideen 211Fig. 2: Fourier transform infrared spectrum of Gracilaria corticataTable 3: The zone of inhibition (mm) of G. corticata against different bacterial pathogensFractions K. pneumonia A. hydrophilla S. aureus E. coli Pseudomonas sp.F1 4±0.50 5±0.52 4±0.50 4±0.62 4±0.42F2 7±0.15 6±0.24 4±0.35 5±0.60 6±0.17F3 5±0.55 4±0.84 5±0.82 4±1.81 5±1.63Crude 8±0.44 7±1.68 6±3.48 6±0.89 9±0K. pneumonia: Klebsiella pneumonia, A. hydrophilla: Aeromonas hydrophilla, S. aureus: Staphylococcus aureus, E. coli: Escherichia coli, G. corticata: Gracilaria corticataTable 4: FT-IR absorption frequencies (cm−1), intensity estimation and functional group of seaweeds G. corticataIR frequency (cm−1)Bond Functional groupsG. corticata3500-3200 O–H stretch, H–bondedAlcohols, phenols3321.463000-2850 C–H stretch Alkanes 2925.492270-1950 X=C=Y Allenes, ketenes, isocyanates, isothiocyanates2084.831670-1600 C=O Stretch Amides 1647.841500-1400 C–C stretch Aromatics 1471.301250-1020 C–N stretch Aliphatic amines 1116.62910-665 N–H wag Primary, secondary amines874.41850-550 C–Cl stretch Alkyl halides 750.98850-550 C–Cl stretch Alkyl halides 712.56690-515 C–Br stretch Alkyl halides 657.10690-515 C–Br stretch Alkyl halides 617.26G. corticata: Gracilaria corticata, FT-IR: Fourier transform infraredACKNOWLEDGMENTSThe authors are thankful to University Grants Commission (UGC), New Delhi for financial assistants and the management of V. O. Chidambaram College, Thoothukudi for providing the necessary facilities, to carry out this work.REFERENCES1. Oh KB, Lee JH, Chung SC, Shin J, Shin HJ, Kim HK, et al. Antimicrobial activities of the bromophenols from the red alga Odonthalia corymbifera and some synthetic derivatives. Bioorg Med Chem Lett 2008;18(1):104-8.2. Soobrattee MA, Neergheen VS, Luximon-Ramma A, Aruoma OI, Bahorun T. Phenolics as potential antioxidant therapeutic agents: Mechanism and actions. Mutat Res 2005;579(1-2):200-13.3. Badea V, Balaban DP, Rapeanu G, Maria C, Badea CF. The antibacterial activity evaluation of Cystoseira barbata biomass and some agents upon bacteria from oropharyngeal cavity. Rom Soc Bio Sci 2009;14:4851-7.4. Rajasekar T, Priyadharshini P, Deivasigamani B, Kumaran S, Edward GG, Sakthivel M, et al. Isolation of bioactive compound from marine seaweeds against fish pathogenic bacteria Vibrio alginolyticus (VA09) and characterisation by FTIR. J Coast Life Med ;1(1):6-13.5. Vijayabaskar P, Shiyamala V. Antibacterial activities of brown marine algae (Sargassum wightii and Turbinaria ornata) from the Gulf of Mannar biosphere reserve. Adv Biol Res 2011;5:99-102.6. Siddhanta AK, Mody KH, Ramavat BK, Chauhan VD, Garg HS, Goel AK, et al. Bioactivity of marine organisms: Part VIII – Screening of some marine flora of western coast of India. Indian J Exp Biol 1997;35(6):638-43.7. Paul G, Yusuf S, Sharma S. Unmasking of the Brugada syndrome phenotype during the acute phase of amiodarone infusion. Circulation 2006;114(11):e489-91.8. Afifah SN, Darah I, Fariza SS, Nordin MK, Aili ZN. Antimicrobial activity of various extracts of a tropical Chlorophyta macroalgae, Halimeda discoidea. J Appl Sci 2010;10:3007-13.9. Oulahal N, Adt I, Mariani C, Carnet-Pantiez A, Notz E, Degraeve P. Examination of wooden shelves used in the ripening of a raw milks mear cheese by FTIR spectroscopy. Food Control 2009;20:658-63.10. Ravikumar S, Ramanathan G, Subhakaran M, Inbaneson SJ. Antimicrobial compounds from marine halophytes for silkworm disease treatment. Int J Med Sci 2009;1(5):184-91.11. Kolanjinathan K, Ganesh P, Govindarajan M. Antibacterial activity of ethanol extracts of seaweeds against fish bacterial pathogens. Eur Rev Med Pharmacol Sci 2009;13(3):173-7.12. Kim SK, Wijesekara I. Development and biological activities of marine derived bioactive peptides: A review. J Funct Foods 2010;2:1-9.Asian J Pharm Clin Res, Vol 8, Issue 2, 2015, 209-212 Radhika and Mohaideen 21213. Lammers K, Arbuckle-Keil G, Dighton J. FT-IR study of the changes in carbohydrate chemistry of three New Jersey pine barrens leaf litters during simulated control burning. Soil Biol Biochem 2009;41:340-7.14. Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12(4):564-82.15. Barros L, Calhelha RC, Vaz JA, Ferreira IC. Antimicrobial activity and bioactive compounds of Portuguese wild edible mushrooms methanolic extracts. Eur Food Res Technol 2007;225:151-6.16. Kostic DA, Velickovic JM, Mitic SS, Mitic MN, Randelovic SS. Phenolic content, and antioxidant and antimicrobial activities of Crataegus oxyacantha L (Rosaceae) fruit extract from Southeast Serbia. Trop J Pharm Res 2012;11(1):117-24.

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FOURIER TRANSFORM INFRARED ANALYSIS OF ULVA LACTUCA AND GRACILARIA CORTICATA AND THEIR EFFECT ON ANTIBACTERIAL ACTIVITY

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