ISSN 2348 – 0319 International Journal of Innovative and Applied Research (2014), Volume 2, Issue (7): 57- 60 Journal home page: http://www.journalijiar.com INTERNATIONAL JOURNAL OF INNOVATIVE AND APPLIED RESEARCH RESEARCH ARTICLE Preliminary in Vitro Phytochemical Screenings of the Barks of Parmentiera cereifera Seem Md. Reyad-ul-ferdous1,2*, Md. Asif Hassan2, Nawfel Abdullah3, Mridul sarker3, Faizul Hafiz3and Tasnim Iffat3 1. Department of Pharmacy, Progati Medical Institute, Dhaka-1207, Bangladesh. 2. Department of Pharmacy, North South University, Dhaka-1229, Bangladesh. 3. Department of Pharmacy, East West University, Dhaka-1219, Bangladesh. ……………………………………………………………………………………………………… Abstract: In present study, the bark and fruit extracts of Parmanteira cereifera were subjected to the Preliminary phytochemical investigation suggested the presence of various potential phytochemicals like flavonoids, saponins, tannins, triterpenoids as well as steroids. The presence of these compounds exhibit potential biological activities such as antioxidant, analgesic, cardioprotective, hepatoprotective, antidiabetics, antidiarrhoeal, anticancer, antiasthmatic activities and so on. This is only preliminary studies to isolate potential compounds which may be use as a lead compound for several biological activities. Key Words: Parmentiera cereifera; phytochemical, saponins, tannins, triterpenoids and steroids. ……………………………………………………………………………………………………… 1. Introduction Greek " Phyto" means plant and phytochemicals are usually related to plant pigments. So, fruits and vegetables that are bright colors - yellow, orange, red, green, blue,and purple-generally contain due to the presence of severals phytochemicals and the most nutrients. We can benefit from all of the phytochemicals and nutrients found in plant foods by eating 5-9 servings of fruits and vegetables a day and eating more whole grains, soy and nuts etc. More than 900 different phytochemicals are discovered from plants. In ancient era plants are extensively used for nutrition and remedies of several diseases. Recent research reported to get your Phytos eat 5-9 servings of colorful fruits and vegetables every day. A phytochemical is a natural bioactive compounds found in plant foods, vegetables, nuts, roots, barks that works as nutrients and dietary fiber to protect against disease such as slow the aging process, cancer, heart disease, stroke, high blood pressure, cataracts, osteoporosis, and urinary tract infections. Pronounced "fight-o-chemicals," phytochemicals fight to protect your health. They can have complementary and overlapping mechanisms of action in the body, including stimulation of t he i m mu ne system, antioxidant effects, modulation of detoxification enzymes, antibacterial andantiviral and modulation of hormone metabolism activities ( Aiyelaagbe and Osamudiamen, 2009) and (Egwaikhide et al., 2007). P. cereifera called Candle tree; is a small tree with rough bark belonging to the family Bignoniaceae. The leaves are oblong and acuminate and the flowers are cauliflorous, nocturnal, white, slightly fragrant and calyx spathaceous. The fruits as well as seeds of this tree are berry smooth, pale yellow, candle-like, pendent, edible and used as fodder source. It is native to Panama and cultivated for ornamental uses in many tropical countries (Van Steenis, 1977; Madulid, 2000). As a part of our continuing studies on medicinal plants in Bangladesh, the organic soluble materials of barks of P. cereifera were evaluated for determining phyto-constituents for the first time (Md. Reyad-ul-ferdous et al., 2012). 2. Materials and Methods 2.1. Plant material Plant materials: The Bark of P. cereifera was collected from Mirpur Botanical Garden, Dhaka, Bangladesh, in November 2011. A voucher specimen collected from Bangladesh National Herbarium, Dhaka, Bangladesh (Accession no.36569). The Bark were washed with water to remove all unwanted plant materials and sand, air dried under light exposure (27°C-30°C for 7 days), pulverized in a mill and stored in an airtight container for further use. The air dried and powdered Bark (500 gm) of P. cereifera was macerated in 2.5 L of methanol for 7 days and then filtered through a cotton plug followed by Whatman filter paper number 1. The extract was concentrated with a rotary evaporator at low temperature (40-45 ºC) and reduced pressure. The concentrated methanolic extract (ME) 57 ISSN 2348 – 0319 International Journal of Innovative and Applied Research (2014), Volume 2, Issue (7): 57- 60 was partitioned by modified Kupchan method (Van Wagenen et al., 1993) and the resultant partitionates i.e., pet- ether (PESF), carbon tetrachloride (CTCSF), chloroform (CSF), and aqueous (AQSF) soluble fractions were used for the experimental processes. 2.2. Preparation of extract The powdered crude material (200 g) was extracted thrice in distilled water (5.5 L; 27°C-30°C) on shaker (Stuart Scientific Orbital Shaker, UK) for 48 hours. The extract was filtered using a Buchner funnel and Whatman No.1 filter paper. The filtrate of aqueous extract obtained was quickly frozen at -40°C and dried for 48 h using a freeze dryer (Savant Refrigerated vapor Trap, RV T41404, USA) to give a yield of 30 g of dry extract. The resulting extract was reconstituted with distilled water to give desired concentrations used in this study. 3. Experimental Procedure 3.1. Phytochemical investigation of the plant extract A small portion of the dry methanolic extract was used for several phytochemical tests. Following phytochemicals investigation was conducted in accordance with the methods with little modifications. In present investigation we found several compounds table-1. Phytochemicals are identified by characteristic color changes (Harborne, 1998 and Egwaikhide et al., 2007). 3.1.2. Anthraquinone (Borntrger’s Test) About 0.5 g of the extract was taken into a dry test tube and 5 mL of chloroform was added and shaken for 5 min. The extract was filtered and the filtrate was shaken with equal volume of 10% ammonia solution. A pink violet or red color in the ammonical layer indicates presence of anthraquinone. 3.1.3. Tannins Small quantity of extract was mixed with distilled water and heated on H2O bath. It was filtered and Ferric chloride was added to the filtrate. A dark green color indicates presence of tannins. 5ml of extract and a few drops of 1% lead acetate were added. A yellow precipitate was formed, indicates the presence of tannins. 3.1.4. Flavonoids About 0.2 g was dissolved in diluted NaOH and HCl was added. A yellow solution that turns colorless indicates the presence of flavonoids. 3.1.4. Saponins About 0.2 g of plant extracts was taken and 5 mL of distilled water was added and then boiled. Frothing persistence shows presence of saponins. 3.1.5. Steroids (Libermann- Burchard Reaction) 1ml plant material in 10 mL chloroform filtered. Two hundred milliliter of acetic anhydride was added to 2 mL filtrate with 2 mL H2SO4. The color changes from violet to blue or green in some samples indicating the presence of steroids.1 ml of the extracts was dissolved in 10ml of chloroform and equal volume of concentrated H2SO4 was addwd by sides of the test tube. The upper layer turn red and H2SO4 layer showed yellow with green fluorescence. Which indicate the presence of steroids compounds in barks of P. cereifera. 3.1.6. Phlobatanins About 0.5 g of plant extract was dissolved in distilled water and filtered. The filtrate was boiled with 2% HCl solution and Red precipitate shows that indicate the presence of phlobatanins. 3.1.7. Terpenoids (Salkowski Method) About 0.5 g of each extract in 2 mL of chloroform filtered. Concentrated H2SO4 carefully added to form a layer. A reddish brown color was formed to show positive results for the presence of terpenoids. 58 ISSN 2348 – 0319 International Journal of Innovative and Applied Research (2014), Volume 2, Issue (7): 57- 60 3.1.8. Triterpenoids 10 mg of the extract was dissolved in 1 ml of chloroform; 1ml of acetic anhydride was added following the addition of 2 ml of conc. H2SO4. Formation of reddish violet colour indicates the presence of triterpenoids. 3.1.9. Cardiac Glycoside About 0.5g of each was treated with 2 mL of glacial acetic acid containing a drop of FeCl3 solution. This was under layered with 1 mL of conc. H2SO4 a brown ring obtained at the interface indicated the presence of de-oxy sugar characteristics of cardenolides. Table-1: Phytochemical investigation of bark of candel tree (P.cereifera) Name of the Test Color Results Anthraquinone (Borntrger’s pink violet or red color - Test) Tannins dark green color - Flavonoids yellow solution that turns colorless + Saponins Frothing persistence + Steroids (Libermann Burchard The upper layer truns red and H2SO4 Reaction) layer showed yellow with green - fluorescence Phlobatanins Red precipitate - Terpenoids (Salkowski Method) Formation of reddish violet colour + Triterpenoids H2SO4. Formation of reddish violet + colour Cardiac Glycoside brown ring obtained - ‘+’ = presence. ‘-’ =absence. 4. Results and discussion In present study, among these test we found several potential phytochemicals such as flavonoids, saponin, Terpenoids, Triterpenoids. Flavonoid acts as hepatoprotective through free radical scavenging activity. This antioxidant properties can slow aging, reduces various biological mutation and anti-inflammatory effect probably by inhibitory on enzyme and metabolism of arachidonic acids. Thrombolytic activity may exhibit due to presence of terpenoids, flavonoids triterpenoids etc as