Screening of Secondary Metabolites in Methanolic Leaf and Bark Extracts of Gardenia Resinifera and Gardenia Latifolia B
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Biosci. Biotech. Res. Comm. Vol. 4, No. 1, June, 2011(23-28) Screening of secondary metabolites in methanolic leaf and bark extracts of Gardenia resinifera and Gardenia latifolia B. Jhansi Lakshmi and *K. Jaganmohanreddy Department of Botany, Kakatiya University, Warangal – 506009 (A.P.), India *Corresponding Author ABSTRACT Secondary metabolites are plant natural products in broad sense cover all the chemical compounds which occurs in nature. These are also known as special metabolites. Secondary metabolites derived as a result of plant metabolism. The research work aims to investigate for screening of different secondary metabolites in methanolic extracts of Gardenia resinifera and Gardenia latifolia. Different chemical tests clearly revealed that these two species found to contain Steroids, Phenolic compounds and Flavonoids in leaf and bark. Key words: Gardenia resinifera, Gardenia latifolia, Secondary metabolites, leaf and bark INTRODUCTION: Medicinal plants are gaining global owing to the fact that matrix or sap at the detached end of ear and stem. This the herbal drugs are cost-effective, easily available and substance known as Dikamali or cumbi-gum. Cumbi most importantly, with negligible side effects. In gum is antispasmodic, expectorant, diaphoretic, addition, plants are valuable source of a wide range of carminative, antihelmintic, relieves constipation, pain secondary metabolites, which are used as treat worms. Gum is antimicrobial, anthelmintic; used in pharmaceuticals, agrochemicals, flavors, fragrances, skin diseases. According to Ayurveda it increases colors, biopesticides and food additives. Over 80% of appetite, astringent to bowels, relieves pain of the approximately 30,000 known natural products are of bronchitis, vomiting and constipation. Gum contains plant origin (Philipson, 1990; Balandrin and Klocke, flavonoids-gardenins, wagonin derivatives, de- 1988). In 1985, 3500 new chemical structures have Metangeretin, nevadensin, hexacosylp-coumarate. been identified and 2600 derived from the higher plants. Worldwide, 121 clinically useful prescription drugs are G. latifolia is a small deciduous tree or large shrub. Root derived from plants (Payne et al., 1991). Even today, used as a remedy for indigestion in children. Fruits used 75% of the world's population relies on plants for in affections of the mammary glands. Pounded pulp is traditional medicine. Plants will continue to provide applied to forehead in fever. Stem and fruit used for novel products as well as chemical models for new stomach pain. Fruit extract is used in treating snake- drugs in the coming centuries (Cox and Balick, 1994). bite, sores of hand and feet, stomach ache and wounds. The advent of chemical analysis and the To treat caries, stem bark crushed and boiled in water is characterization of molecular structures have helped in applied to affected areas. Bark is used in skin diseases. precisely identifying these plants and correlating them The bark and wood gave beta sestorol, hederegenin, with their activity under controlled experimentation. Me-esters of oleanic and gypsogenic acids. Root gave Despite advancements in synthetic chemistry, there is gardenins. Saponins from bark decreased formation of need of biological sources for a number of secondary histamine and may find use in asthma (market drug is expectorant and weak spasmolytic, but was not found metabolites including pharmaceuticals (Pezzuto, 1995). effective in asthma). The stem bark contains and Akinpelu and Okoya (2006). hederagenin, D-mannitol, sitosterol and siaresinolic, episiaresinolic, oleanolic andspinosic acid (Moorthy et. Rubiaceous plants elaborate a wide range of chemical al, 2007) substances. This family is well known since time immemorial, as a source of alkaloids, which form the MATERIAL AND METHODS major bulk of chemical substances investigated so far. Plant material: The plant material of G.resinifera and G.latifolia were collected from Kakatiya arboretum Gardenia resinifera Roth. and Gardenia latifolia Ait. are nearer to Kakatiya University, Warangal, A.P. related to Rubiaceae family, which parades a long list of Methanolic Extraction: Plant extracts were prepared by plants of medicinal importance. G.resinifera is shrub or cold percolation method. The plant materials were dried small tree. It bears small leaves and secretes gummy under shade and ground into fine powder using electric Received 10th Feb. accepted after revision 20th June 2011 bender. 50g of dried powder was soaked in 300ml BBRC ISSN : 0974 - 6455 A Soc Sci Nat India Publication 23 Lakshmi and Jaganmohanreddy Biosci. Biotech. Res. Comm. Vol. 4, No. 1, June, 2011(23-28) hexane for 48h with intermittent shaking. The plant presence of these secondary metabolites. Extracts extracts were filtered through whatman No.1 filter paper were tested negative for Alkaloids and Saponins. into pill vials. The filtrates were dried until a constant dry weight of each extract was obtained. The residues were Plants are known to produce secondary metabolites stored at 40 C for further use. The remaining plant that complement their structural barrier in their efforts residue was dried and soaked in 300ml of methanol as (De Lucca et al., 2005). A number of secondary above and the extract was collected as described earlier metabolites are associated with different potentials in (Nair et al, 2005). various plants (Ojekale et al., 2006 ; Smith et al., 2001 ; Cowan,1999 ; Huynch et al., 1996). Several researchers Secondary Metabolite Screening contributed similar type of investigations in the different The qualitative phytochemical analysis of G.resinifera plant species namely Grewia tilifolia, Tridax and G.latifolia was tested as follow. procumbens, Senna uniflora and Dysoxylum species Chemical Tests (Badani et al., 2002 ; Suseela et al., 2002 ;Vijai et al., I) Test for Alkaloids: A small quantity of methanolic 2004 : Parcha et al., 2004). On the otherhand, Aggarwal extract of the plant species was taken and it was et al., (2002) reported various chemical constituent from evaporated to dryness. The residue was then dissolved Achyranthes species (Achyranthes aspera, A.bidentata in 1% HCl and tested with Mayer's and Dragen dorff's and fauriei) including steroids, long chain compounds, reagents. When a precipitate forms or when the solution saponins, organic acids and their esters, become turbid, the reaction was considered as positive carbohydrates, alkaloids, triterpenes, polyphenols, for the presence of alkaloids. anthraquinones, flavonoids and amino acids. II) Test for Steroids: In both species of Phyllanthus burchellii and A) Leibermann-burchard's test: Methanolic extract Phyllanthus pavulus are rich in alkloids and saponins. were treated with 50% sulphuric acid and a few drops of However, in the P.pavulus extracts tannin, flavonoids acetic anhydride. The development of reddish brown and anthraquinones were in appreciable amounts. ring indicates the positive reaction of steroids. Anthocynoides was absent in this species (Mdlolo et al., B) Salkowski test: To 1ml of methanolic extract a few 2008). These secondary metabolites are known to drops of sulphuric acid and chloroform were added. The exhibit medicinal activity as well as physiological development of wine red colour indicates the presence activity. The phytochemical screening revealed the of steroidal nuclei. presence of saponins, cardenolides and tannins in the leaf of Flabellaria paniculata while anthrquinones, III) Test for flavonoides cyanogenic glycoside and flavonoides were found A) Flavonoides test: 1ml of methanolic algal extracts a absent (Abo and Olugbuyiro, 2004). few drops of sulphuric acid and magnesium turnings were added. The development of pink or magenta Parkia bicolor and Parkia biglobosa contained cardiac colour, scarlet colour and deep cherry red colour glycosides, though it gave a much more positive test for indicates the positive reaction for Flavonols, Flavones that of P.biglobosa. Tannins were also indicated in the and Flavonones respectively. two leaf extracts. There was complete absence of B) Shinoda test: 1ml of methanolic algal extract pinch saponins and anthraquinones in both extracts of sodium hydroxide and few drops of sulphuric acid (Ajaiyeoba and Edith, 2002). In Mallotus philippinensis were added. Development of yellow brown colour hexane extract revealed the presence of phenolic indicates the positive reaction for Flavonoides. groups, chloroform extract showed the presence of alkaloids, phenolic groups, steroids and methanol IV) Test for Phenolic compounds- Small quantities of extract showed the presence of flavones, phenolic alcoholic and aqueous extracts in water are tested for groups, saponins, steroids, sugars, tannins and the presence of phenolic compounds with dilute ferric triterpenes (Moorthy et al., 2007). Psidium guajava chloride solution (5%), 1% solution of gelatin containing methanolic extracts revealed the presence of tannins 10 % sodium chloride, 10% lead acetate and bromine while that of Mangifera indica showed the presence of solutions. alkaloids, saponinss and tannins (Akinpelu and Onakoya, 2006). These compounds are known to be RESULTS AND DISCUSSION: biologically active. Tannins have been found to form The chemical constituents or phytochemicals of the leaf irreversible complexes with proline-rich proteins and bark extracts of G.resinifera and G.latifolia were resulting in the inhibition of the cell protein synthesis. identified by qualitative chemical tests. Phytochemical screening revealed the presence