STUDIES ON GC-MS PROFILING OF SOME SEAWEEDS OF MAHIM BEACH DIST. PALGHAR, MAHARASHTRA USING VARIOUS SOLVENTS

STUDIES ON GC-MS PROFILING OF SOME SEAWEEDS OF MAHIM BEACH DIST. PALGHAR, MAHARASHTRA USING VARIOUS SOLVENTS

Viraj Chabake ¹, Sakshi Chaubal ^{* 2} and Tanmay Patil ¹

Department of Botany ¹, The Institute of Science, Mumbai - 400032, Maharashtra, India.

Department of Botany ², Mah. Govt’s Elphinstone College Mumbai - 400032, Maharashtra, India.

ABSTRACT: Objective: To investigate the bioactive constituents of Ulva lactuca, Sargassum prismaticum, and Sarconema filiforme using gas chromatography-mass spectrometry (GC-MS) analysis. Methods: 10 gm sample of each seaweed was successfully extracted with HPLC grade methanol and pet. ether with continuous shaking in a dark room for 36 h. The concentrated extracts were subjected to GC MS analysis. Results: The GC MS analysis of methanol and pet ether extracts of U. lactuca, S. prismaticum and S. filiforme show various types of phytocompounds screened under different R.T. Major compounds were 9-Octadecenoic acid (Z)-, phenyl-methyl ester, Stannane,1,3-dithian-2-ylidenebis[tributyl,Octadecane, 3-ethyl-5-(2-ethylbutyl) in methanol extract and Decane, 6-ethyl-2-methyl-, Tricosane and Sulfurous acid, 2-ethylhexyl hexyl ester in pet. ether. These bioactive compounds are considered as biologically and pharmacologically important. Conclusion: The three different extracts contain diverse bioactive compounds that were identified and characterized spectroscopically. Hence, the identification of different biologically active compounds in the extracts of seaweeds justifies further biological and pharmacological studies.

Bioactive compounds, Crude extract, GC-MS analysis, Seaweeds etc.

INTRODUCTION: Seaweeds are one of the most significant existing marine resources in the world. These macroalgae have been a good source of food, fodder, manure, and medicine in the east as well as in the west, since centuries ^{1, 2}. Seaweeds have been classified into three major divisions, namely Chlorophyta (Green algae), Phaeophyta (Brown algae), and Rhodophyta (red algae) based on their photosynthetic pigments and reserve food ³. Seaweeds are a significant marine resource as they are able to produce a great variety of secondary metabolites that could lead to the development of new functional ingredients ⁴.

Seaweeds comprise of large number of structurally diverse bioactive compounds such as alkaloids, glycosides, saponins, tannins, flavonoids, steroids etc., which are of immense medicinal value and use in a broad spectrum of biological activities ⁵.

All over the world, seaweeds have been studied and found to have enormous therapeutic potential as they possess anti-oxidant, neuroprotective, anti-fungal, anti-viral anti-inflammatory, anticancer, anti-HIV, antimutagenic, scavenging free radicals and antimicrobial activities. Seaweeds are also good source of vitamins such as A, B_l, B₁₂, C, D, E, riboflavin, niacin, pantothenic acid and folic acid as well as minerals such as Ca, P, Na, and K ⁶. Variation in nutrient content and bioactive compounds is observed in seaweeds in different seasonal cycles and with different environmental factors like ecological distribution, salinity, temperature, light, etc. ^{7, 8, 9, 10}

In spite of their significance towards mankind, seaweeds are relatively ignored in India. India has a total geographical area of about 329 million hectares with a vast coastline of over 7000 km ^{11, 12}. Along the coast of India, many of the rocky beaches, coral reefs, lagoons, estuaries, and mudflats provide ideal habitats for the growth of marine algae ¹³. The marine algae resources are abundant around coastlines of Mumbai, Palghar, Raigad, Ratnagiri, and Sindhudurg in Maharashtra ¹⁴. The distribution of seaweed resources along the Indian coast was first mapped by Thivy ¹⁵.

Large numbers of bioactive compounds have been isolated from the seaweeds so far; still, there is a need to explore and analyze many more seaweeds of different places which might show variation in biochemical content and could be used to produce various food supplements, industrial products, manure and compounds of pharmaceutical interest to cure different ailments.

Mahim beach, Dist. Palghar is a rocky beach. It shows ample collection of several seaweeds. Despite wealth of valuable raw products, seaweeds of this beach remained largely unexplored. Ulva lactuca, Sargassum prismaticum, and Sarconema filiforme were abundant at the Mahim beach and thus were selected for the present study of some bioactive compounds contained in them. Ulva lactuca belonging to the division Chlorophyta is a potential source of polysaccharides, proteins, pigments, cellulose, amino acids, lipids, fatty acids, and minerals, etc. ^{16, 17} It has the capacity to rapidly proliferate and stated to exhibit a wide range of applications in food and health industry. Significant therapeutic properties like antibacterial, antifungal, antioxidant etc., are reported from Ulva sp. collected all over the world. Sarconema filiforme belonging to division Rhodohyta has considerable pharmacological, antimicrobial, antifungal, and cytotoxic potentials ¹⁸.

Few sps. of Sargassum (Phaeophyta) are reported to be good source of phytochemical contents such as carotenoids, terpenes, phenolic compounds, fatty acids, lipids, and amino acids ¹⁹. GC MS technique was selected for the characterization as it is the most appropriate technique to analyze and identify bioactive components of alcohols, esters, fatty acids, amino acids, steroids, long-chain hydro-carbons, essential oils, etc. Crude seaweed extract was made using solvents of different polarities.

Studies related to bioactive compounds of seaweeds of Mahim beach, Dist Palghar are reported for the first time.

MATERIALS AND METHODS:

Collection of Seaweeds: Fresh seaweed Ulva lactuca, Sargassum prismaticum, and Sarconema filiforme were collected from intertidal regions of Mahim beach, dist. Palghar during the month of December 2017. The collected seaweed sample was cleaned with the seawater until unwanted impurities and adhering sand particles were removed. It was then cleaned thoroughly with tap water to remove the salty surface materials. The seaweed sample was shade dried for 7-8 days. The dried seaweed samples were powdered using mixer and passed through 0.5 mm sieve. It was stored in refrigerator for further study.

Seaweed Extraction: Seaweed extracts were obtained using solvents of different polarities. The seaweed powdered sample (10 gm) was extracted successively with HPLC grade methanol and pet ether using the cold extraction method. The sample was kept in a dark room for 36 hrs with continuous shaking. The extract was collected and filtered using Whatman no.1. It was evaporated to dryness by a rotavap. The final residue obtained was stored at 4 ºC until further use. The volatile bioactive compounds present in methanol and pet. ether extracts of the seaweeds were identified by GC-MS characterization.

GC-MS Instruments: GC MS analysis of a different solvent extract of Ulva lactuca, Sargassum prismaticum, and Sarconema filiforme was carried out with Agilant 7890 system and gas chromatography interfaced to a mass spectrometer (GC-MS) employing the following conditions: HP 5 column [(5%-Phenyl)-methylpolysiloxane) (30mm × 0.25mm × 0.25μm] thickness, Helium gas (99.999%) was used as a carrier gas at a constant flow of 1ml/min, and injection volume of 1μl was employed (split ratio of 10:1) operating in electron impact mode at 70eV; injector temperature 250 ºC; Ion-source temperature 280 ºC. The oven temperature was programmed from 80 ºC (isothermal for 1 min), with an increase of 10 ºC /min, to 200 ºC then 5 ºC/min to 280 ºC ending with 9 min, isothermal at 280 ºC. Mass spectra were taken at 70eV; a scan-interval of 0.5 seconds and fragments vary from 45-550 Da. Total GC running time was 35 min.

Identification of Chemical Constituents: The resulting peaks were analyzed with the database of the National Institute Standard and Technology (NIST) a library, which has more than 62,000 patterns. The spectrum of the unknown component was compared with the spectrum of the known components stored in it. The Name, Molecular weight and structure of the components of the seaweed sample was determined ²⁰.

RESULTS: The volatile bioactive compounds present in methanol and pet. ether extract of Ulva lactuca, Sargassum prismaticum, and Sarconema filiforme were identified by GC-MS analysis. The methanol and pet. ether extracts of seaweeds indicated the presence of different types of phytocompounds Table 1-2 screened under different retention times (R.T). A total of (9, 4 and 9 of methanol extract) and (4, 7, and 11 of pet. ether extract) were observed with retention times for U. lactuca, S. prismaticum and S. filiforme respectively, as shown in Fig. 1–6. Chemical constituents were identified using spectral database NIST 11 software installed in the GC–MS. The compounds prediction is based on Pubchem ²⁰. The compound name with R.T, molecular formula, molecular weight and concentration % in the methanol and petether extract of seaweeds Ulva lactuca, Sargassum prismaticum and Sarconema filiforme are presented in Table 1-2. 9-Octadecenoic acid (Z)-, phenylmethyl ester are most abundant (R.T 31.81, 31.52%) in methanol U. lactuca extract followed by Stannane, 1,3-dithian-2-ylidenebis[tributyl- (R.T 14.82, 70.59%) in S. prismaticum and Octadecane, 3-ethyl-5-(2-ethylbutyl)- (R.T 31.62, 36.50%) present in S. filiforme. Crude extract of petetherindicates most abundant compound as Decane, 6-ethyl-2-methyl- (R.T 31.86, 41.07%), Tricosane  (R.T 31.66, 25.84 %) and Sulfurous acid, 2-ethylhexyl hexyl ester (R.T 19.42, 31.96%) respectively.

TABLE 1: COMPOUNDS IDENTIFIED IN THE METHANOL EXTRACT OF ULVA LACTUCA, SARGASSUM PRISMATICUM AND SARCONEMA FILIFORME BY GC-MS ANALYSIS

TABLE 2: COMPOUNDS IDENTIFIED IN THE PET. ETHER EXTRACT OF ULVA LACTUCA, SARGASSUM PRISMATICUM AND SARCONEMA FILIFORME BY GC-MS ANALYSIS

FIG. 1: GC–MS CHROMATOGRAM OF THE CRUDE METHANOL EXTRACT OF ULVA LACTUCA

FIG. 2: GC–MS CHROMATOGRAM OF THE CRUDE METHANOL EXTRACT OF SARGASSUM PRISMATICUM

FIG. 3: GC–MS CHROMATOGRAM OF THE CRUDE METHANOL EXTRACT OF SARCONEMA FILIFORME

FIG. 4: GC–MS CHROMATOGRAM OF THE CRUDE PET. ETHER EXTRACT OF ULVA LACTUCA

FIG. 5: GC–MS CHROMATOGRAM OF THE CRUDE PET. ETHER EXTRACT OF SARGASSUM PRISMATICUM

FIG. 6: GC–MS CHROMATOGRAM OF THE CRUDE PET. ETHER EXTRACT OF SARCONEMA FILIFORME

DISCUSSION: Marine algae have been rich source of carotenoids, polyunsaturated fatty acids ²¹, polysaccharides ²², flavonoids, phenolic acids ²³, sterols ²⁴ Phlorotannins ²⁵ etc. Present GC–MS analysis of crude extracts of methanol and pet. ether of Ulva lactuca, Sargassum prismaticum and Sarconema filiforme revealed various compounds. Major compounds were 9-Octadecenoic acid(Z)-, phenylmethyl ester, Stannane, 1,3-dithian-2-ylidenebis [tributyl, Octadecane, 3-ethyl-5-(2-ethylbutyl) in methanol extract and Decane, 6-ethyl-2-methyl-, Tricosane and Sulfurous acid, 2-ethylhexyl hexyl ester in pet. ether. Some other important bioactive compounds mentioned in Table 1-2 are rich source of structurally novel and biologically active metabolites and can be used as potential natural antioxidants in different food and pharmaceutical products ²⁶. Babu et al., (2014) reported 17 different components in ethanolic extract of Ulva lactuca. Major components Phytol (10.51%), Hexadecanoic acid, ethyl ester (20.24%) and (E)-9- Octadecenoic acid ethyl ester (9.59%) are considered to have significant medicinal properties. Kumar et al., (2013) found 12 compounds having antibacterial activity in methanolic extract of Sargassum tenerrimum by GC MS analysis. Red algae Halimeda opuntia and Sarconema filiforme are described as potent algal species with considerable antimicrobial, anti-plasmid, and cytotoxic activity ²⁹.

CONCLUSION: The present study helps to predict the formula and structure of bioactive compounds in Ulva lactuca, Sargassum prismaticum, and Sarconema filiforme. Further work related to verifying their efficacy may lead to the improvement of drug formulation.

ACKNOWLEDGEMENT: Nil

CONFLICTS OF INTEREST: Nil

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    How to cite this article:

    Chabake V, Chaubal S and Patil T: Studies on gc-ms profiling of some seaweeds of Mahim Beach Dist. Palghar, Maharashtra using various solvents. Int J Pharm Sci & Res 2021; 12(3): 1644-50. doi: 10.13040/IJPSR.0975-8232.12(3).1644-50.

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