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3556 Journal of Pharmacognosy and Phytochemistry 2019 83: 3556-3559 E-ISSN: 2278-4136 P-ISSN: 2349-8234 JPP 2019 83: 3556-3559 Received: 13-03-2019 Accepted: 15-04-2019 V Smitha Research Scholar PG Research Department of Botany V.O. Chidambaram College Tuticorin Tamil Nadu India V Vadivel PG and Research Department of Botany V. O. Chidambaram College Tuticorin Tamil Nadu India Correspondence V Vadivel PG and Research Department of Botany V. O. Chidambaram College Tuticorin Tamil Nadu India Phytochemical screening for active compounds in Ceratopteris thalictroides L. Brogn V Smitha and V Vadivel Abstract The plant whole Ceratopteris thalictroides L. Brogn collected from Puthalam Kanyakumari District Tamil Nadu India were analyzed for the presence of different phytochemicals. The aim of our study is to screen the biologically active compounds in plant material C. thalictroides. Phytochemical methods of screening proven the presence of alkaloids steroids coumarin tannins saponins flavonoids quinone anthroquinone phenol protein xanthoprotein carbohydrate glycosides catachin sugar and terpenoids in the extracts of the whole plants. The phytochemical composition of the whole plants indicate their medicinal properties. Keywords: Phytochemical pteridophyte Ceratopteris thalictroides Introduction Like angiosperms plants phytochemical studies of fern plants do not worked out extensively. From phytochemical analyses it has been observed that fern plants contain higher levels of carbohydrate amino acids protein lipid and secondary metabolites and these are economically useful product of any region 1 . Phytochemical characterization of plant material is important as it relates to the therapeutic actions. It is perhaps obvious that different species of plants would have different chemical constituents. Phytochemicals are non-nutritive plant chemicals that have protective or disease preventive properties. Phytochemicals are organic non-nutritive naturally occurring chemicals found in plant foods. They are nonessential nutrients meaning that they are not required by the human body for sustaining life. It is well-known that plant produces these chemicals to protect them but recent research demonstrates that they can also protect humans against diseases. Further the importance of secondary metabolites like phenolic compounds and alkaloids as medicinal value has been highlighted. Several studies 2 3 4 5 were made to evaluate the importance of ferns from chemical and pharmacological aspects. Humans use secondary metabolites as medicines flavourings and recreational drugs. Ceratopteris thalictroides occurs in semi shaded localities mostly rooted in mud occasionally free floating and common in paddy fields ponds 6 7 8 . The fronds of C. thalictroides are used as a vegetable 9 10 . The fronds of C. thalictroides are used as poultice in skin diseases 11 . The uncurled fronds are eaten as a salad or as a substitute for asparagus. The tribal people use the plant as a poultice for skin problems 12 . The whole plant parts are ground into paste and mixed with turmeric. The mixture is applied over the affected places to treat cure skin diseases and wounds 13 14 . In Madagascar C. thalictroides leaves are eaten as salad or cooked as vegetable whereas in Swaziland leaves are eaten as leafy vegetable 15 . Materials and Methods Plant Material The plant material Ceratopteris thalictroides L. Brogn. was collected from Puthalam 8.106488 77.46 Kanyakumari District Tamil Nadu India and was authenticated at Botanical Survey of India Southern circle Coimbatore Tamil Nadu a voucher specimen VS- VV-01 has been deposited in the Department of Botany V.O. Chidambaram College Tuticorin. Preparation of Plant Extract The collected plant material was washed under running tap water followed by sterilized distilled water to remove the soil and dust particles. The damaged parts of the plant was removed cut into small pieces and then shade dried. The shade dried plant pieces were ground to coarse powder with the help of an electric grinder. After that the powdered plant material was stored in air-tight container for further investigation.

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3557 Journal of Pharmacognosy and Phytochemistry The coarse powder was subjected to extraction in 250ml each of petroleum ether benzene chloroform ethyl acetate ethanol and methanol solvents separately. The coarse powder 10g of the plant material was weighted and put into the brown glass bottles. Then the solvents were added to it. Then the bottles were sealed with aluminium foil and kept in laboratory shaker at room temperature and the bottles were shaken for one week. Finally the extract was filtered through many layers of muslin cloth for coarse filtration. The coarse filtrate was than filtered through Whatman number 1 filter paper. The obtained filtrate was evaporated in a vacuum rotary evaporator under reduced pressure at 40ºC until the filtrate was reduced to one-third of the starting filtrate volume and the concentrated extracts were further evaporated to get dry extracts. A part of dry extracts were re-dissolved in dimethyl sufloxide DMSO and were stored in stopper glass bottles and another part was kept as such in air-tight bottles at 0ºC for further analysis. Phytochemical Screening The different solvent extracts of Ceratopteris thalictroides were used for screening the presence of alkaloids steroids coumarin tannins saponins flavonoids quinone anthroquinone phenol protein xanthoprotein carbohydrate glycosides catachin sugar and terpenoids according to standard procedures of Harborne 16 Brindha et al. 17 Trease and Evans 18 and Sofowara 19 . Screening for Alkaloids Dragendroff’s test 2ml of the extract was mixed with 8ml of 1 HCl warmed and filtered. Then the filtrates were treated with Dragendroff’s reagent solution of Potassium Bismuth Iodide. Formation of red precipitate indicates the presence of alkaloids. Screening for Steroids Liebermann Burchard test Extracts were treated with chloroform and filtered. The filtrates were treated with few drops of acetic anhydride boiled and cooled. Concentrated sulphuric acid was added. Formation of brown ring at the junction indicates the presence of phytosterols. Screening for Coumarin 2ml of the extracts was taken in test tubes. The mouth of the tube was covered with filter paper treated with 3ml of 1N NaOH solution. Test tube was placed for few minutes in boiling water and then the filter paper was removed and examined under the UV light for yellow fluorescence indicated the presence of coumarins. Screening for Tannins 50mg of various solvent extract powder was dissolved in 10ml distilled water and filtered. 1 aqueous iron chloride FeCl 3 solution was added to the filtrate. The appearance of intense green purple blue or black colour indicated the presence of tannins in the test samples. Screening for Saponin 50mg of the various solvent extract powder was boiled in distilled water in a test tube in boiling water bath and filtered. 10ml of the filtrate was mixed with 5ml of distilled water and was shaken vigorously to the formation of stable persistent froth. The frothing was mixed with 3 drops of olive oil and shaken vigorously for the formation of emulsion thus a characteristic of saponins. Screening for Flavonoids Shinoda Test To the extract solution 5ml added few fragments of magnesium ribbon and concentrated HCl drop wise. Appearance of red or orange red colour indicates the presence of flavonoids. Screening for Quinone 1ml of the extract was mixed with 1ml of concentrated H 2 SO 4 . Appearance of red colour shows the presence of Quinone. Screening for Anthroquinone Borntrager’s test 50mg of extract powder was taken into a dry test tube and 5ml of chloroform was added and shaken for 5min. The extract was filtered through Whatman No 1 filter paper and the filtrate was shaken with equal volume of 10 ammonia solution. A pink violet or red colour in the ammoniacal layer lower layer indicates the presence of anthroquinone. Screening for Phenols The extract powder 50mg was dissolved in 5 ml of distilled water. To this few drops of 10 ferric chloride solution was added. Appearance of blue or green colour indicates the presence of phenol compounds. Screening for Protein The extract powder 50mg was dissolved in 10ml of distilled water and filtered through Whatman No. 1 filter paper. To the filtrate 1ml of 40 NaOH was added. Then 1 or 2 drops of 2 copper sulfate solution was added. Appearance of violet colour indicates the presence of proteins. Screening for Xanthoprotein One ml each of the various extracts were treated separately with few drops of concentrated HNO 3 and NH 3 solution. Formation of reddish orange precipitate indicates the presence of xanthoproteins. Screening for Carbohydrates Molisch Test To 2ml of extracts 3 drops of α-naphthol 20 in ethanol was added. Then 1ml of concentrated sulphuric acid was added along the side of the test tube. Reddish-violet ring at the junction of the two layers indicated the presence of carbohydrates. Screening for Glycosides Borntrager’s test Extract powder 50mg was mixed with concentrated H 2 SO 4 5ml. then it was heated for 3 minutes thereafter it was filtered after that filtrate was mixed with 0.5ml of 10 NaOH and allowed to stand for 3 minutes. Appearance of reddish brown precipitate indicates the presence of glycosides. Screening for Catachin To the extracts a few drops of Ehrlich’s reagent and concentrated hydrochloric acid were added. Appearance of pink colour indicates the presence of catechin. Screening for Reducing Sugar For the presence of reducing sugars in the extract Fehling test was performed. An amount of 50mg of the extract powder was taken and added it to the equal volume of boiling Fehling solutions A and B in a test tube. A brick- red precipitates indicates the presence of reducing sugar

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3558 Journal of Pharmacognosy and Phytochemistry Screening for Terpenoids Salkowski test 5ml of various solvent extract was mixed in 2 ml of chloroform followed by the careful addition of 3ml concentrated sulfuric acid H 2 SO 4 . A layer of the reddish brown colouration was formed at the interface thus indicating a positive result for the presence of terpenoids. Result and Discussion Preliminary phytochemical screening of plants is important in the detection of bioactive principles which is a new source of therapeutically and industrially valuable compounds that may lead to the discovery of new drugs. In the present study the presence of sixteen phytochemicals were screened in the petroleum ether benzene chloroform ethyl acetate ethanol and methanol extracts of the whole plants of Ceratopteris thalictroides and the results are shown in Table 1. Table 1: Phytochemical compounds detected in the plant extracts. S. No. Compounds Petroleum Ether Benzene Chloroform Ethyl Acetate Ethanol Methanol 1 Alkaloids - - + + + + 2 Steroids + + + + + + 3 Coumarin - + + + - + 4 Tannins - - + - - + 5 Saponins + + - + + + 6 Flavonoids + + - - - + 7 Quinone + + + + - + 8 Anthroquinone + - + - + - 9 Phenol + + + + + + 10 Protein - + + - - + 11 Xanthoprotein - + + - - + 12 Carbohydrate + + - + + - 13 Glycosides + - - + + + 14 Catachin + + + + - + 15 Sugar + + - - - - 16 Terpenoids - - + - - + Presence or absence of certain important bioactive compounds in an extract is determined by colour reactions of the compounds with specific chemicals which act as dyes. This procedure is a simple preliminary pre-requisite before going for detailed phytochemical investigation. In India traditional communities like tribal and rural populations are frequently using the crude extracts of local plants for medicinal and other purposes. Crude extracts and medicines manufactured on the principles of natural compounds even by pharmaceutical companies may lead to large scale exposure of humans to natural products. The first step towards this goal is the biological and phytochemical screening of plant extracts from traditional preparations used in popular medicine. Hence in the present study the crude extracts obtained by petroleum ether benzene chloroform ethyl acetate ethanol and methanol solvents were screened for the presence of phytochemicals. The petroleum ether extract showed the presence of steroids saponins flavonoids quinine anthroquinone phenol carbohydrate glycosides catechin and reducing sugar. The benzene extract showed the presence of steroids coumarin saponins flavonoids quinine phenols protein xanthoprotein carbohydrate catechin and reducing sugar. The chloroform extract showed the presence of alkaloids steroids coumarin tannins quinine anthraquinone phenol protein xanthoprotein catechin and terpenoids. The ethyl acetate extract showed the presence of alkaloids steroids coumarin saponins quinon phenol carbohydrate glycoside and catechin. The ethanol extract showed the presence of alkaloids steroids anthroquinone phenols carbohydrate and glycosides. The methanol extract showed the presence of alkaloids steroids coumarin tannins saponins flavonoids quinone phenol protein xanthoprotein glycosides catechin and reducing sugar. Among the phytochemicals steroids and phenol were detected in all the presently investigated solvent extracts. This research findings highlights that due to the abundance of various phytochemicals in the Ceratopteris thalictroides methanolic plant extract it holds the great potential to treat various human diseases and has profound medical applicability. The presence of these important phytochemicals in C. thalictroides signals their therapeutic potential. They may function in stimulating digestion act as anti- inflammatory reducing swelling and pain antioxidant and venotonics antibacterial and antifungal diuretic property that enhance the elimination of waste products and toxins and enhancing mood to give a sense of well-being 20 . Besides the present study was also conducted with an objective to identify the best extraction solvent which can be used to extract the maximum amount of the phytochemicals from the Ceratopteris thalictroides dried plant leaves. Among all the solvents the highest number of compounds were detected in methanol extract 13 compounds. Conclusion A comparative study has been conducted with an aim to achieve the best extraction solvent for the extraction of phytochemicals from Ceratopteris thalictroides plant. The results from this study demonstrate that using methanol as extraction solvent results in the maximum extraction of phytochemicals. Besides Ceratopteris thalictroides have known to possess many phytochemicals which play an important role in fighting against pathogens. The phytochemical screening demonstrated the presence of the secondary metabolites i.e. alkaloids steroids coumarin tannins saponins flavonoids quinone anthroquinone phenol glycosides catachin and terpenoids in extracts of Ceratopteris thalictroides. The phytochemical analysis of the plants is also important in pharmaceuticals companies for the novel drugs for the treatment of various diseases. Further studies are needed to strengthen the disease fighting ability of the Ceratopteris thalictroides.

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