Meenaa Venkataramani et al. / Journal of Pharmacy Research 2012,5(5),2895-2899 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Preliminary phytochemical screening and GC-MS profiling of Hiptage benghalensis (L.)kurz Meenaa Venkataramani* and Sasikumar Chinnagounder1 *PG and Research Department of Biotechnology, Nehru Memorial College (Autonomous), Puthanampatti, Tiruchirapalli, Tamilnadu. India 1Head, PG and Research Department of Biotechnology, Nehru Memorial College (Autonomous), Puthanampatti, Tiruchirapalli, Tamilnadu. India Received on:11-02-2012; Revised on: 17-03-2012; Accepted on:19-04-2012 ABSTRACT The present study is focused on the preliminary phytochemical screening of various leaf extracts of Hiptage benghalensis and also in analyzing the phytocomponents using Gas chromatography-Mass Spectroscopy. The preliminary phytochemical screening revealed the presence of Coumarins, Sterols, Lignin, Saponins, Flavanoids, Alkaloids, Tannins, Terpenes and Protein in various leaf extracts of the plant under study and the GC-MS investigation of the chloroform fraction of the plant retrieved a total of 55 components which have been reported and discussed below. Keywords: Phytochemical screening, Gas Chromatography–Mass Spectroscopy, Hiptage benghalensis. INTRODUCTION The Indian subcontinent is a vast repository of medicinal plants that are used also to quench thirst. According to some researches the therapeutic actions of in traditional medical treatments(1).Chemical principles from natural sources this plant may be due to the presence of mangiferin (13), which is known to be have become much simpler and have contributed significantly to the develop- anti-inflammatory, hepatoprotective, antioxidant, and antimicrobial (14),(15). ment of new drugs from medicinal plants (2-3). Biologically active compounds The pharmacological potentials of H.benghalensis Kurz and the various from natural sources have always been of great interest to scientists working phytocomponents attributing to it still remain unexplored. Hence the present on infectious diseases. Research to find out scientific evidence for claims of investigation focuses on the preliminary phytochemical screening of aque- plants used for Indian Ayurvedic system of medicine has been intensified. In ous, ethanol, methanol and chloroform leaf extracts of H.benghalensis col- recent years the thirst to investigate on the bioactive components of the lected from Kolli hills, Tamilnadu and analysis of the phytoconstituents by plants has been incredibly focused on. Chromatographic separation with Gas chromatography and Mass Spectroscopy. mass spectrometry for the chemical characterization and composition analy- sis of botanicals has been growing rapidly in popularity in recent years. MATERIALS AND METHODS: Even though global herbal resources have a great potential as natural drugs Plant Material and are of great commercial importance, they are very often procured and Aerial parts of the selected plant were collected from the Eastern Ghats, processed without any scientific evaluation, and launched onto the market Tamilnadu. The taxonomic identification and authentication of the plant was without any mandatory safety and toxicology studies (4-5). The lack of phar- certified (PARC/2012/1238) by Prof.Dr.P.Jayaraman, Director, Institute of macological and clinical data on the majority of herbal medicinal products is Herbal Botany, Plant Anatomy Research Centre, Chennai. a major impediment to the integration of herbal medicines into conventional medicinal practice. For valid integration, pharmacological studies must be Extraction of Plant material conducted on those plants lacking such data (6,7,8). The list of many such The collected leaves were dried, coarsely powdered using electrical grinder undocumented, unexploited and uncharacterized plants of medical impor- and were subjected to cold and hot extractions. A definite quantity of the tance have been brought to light by many researchers and one such unex- powder was weighed (1:3) and soaked in the respective solvents for 2 days at plored plant is Hiptage benghalensis. room temperature with intermittent shaking and on the 3rd day, extracts were separated using Whatman No.1 filter paper. The hot extraction of the plant Hiptage benghalensis (L) Kurz. belongs to the family Malphigiaceae. It is a was carried over subsequently and with sufficient volume of various organic large, woody climber found in India, Burma, Malaysia, Thailand, China and solvents in the order of increasing polarity. The collected extracts were used the Philippines (9-11). The bark, leaves and flower of H.benghalensis are aro- for further analysis. All chemicals and solvents used for different studies matic. They are useful in conditions of burning sensation, wounds, ulcers, were of analytical grade. inflammations, leprosy, scabies, cough and rheumatism (12). The plant has tremendous therapeutic potential with every part of the plant being used Preliminary Phytochemical Screening of various extracts medicinally. The leaves of H.benghalensis (L.) Kurz are used in treating skin Preliminary phytochemical screening of various extracts was carried out as diseases in Burma and the bark is used to heal wounds in Indonesia. In India, per the standard textual procedure (16). H.benghalensis (L.) Kurz is widely used to treat cough, asthma, leprosy and Test for Saponins: *Corresponding author. The substance was shaken well with water. Meenaa Venkataramani Test for Tannins: PG and Research Department of Biotechnology, The substance was mixed with basic lead acetate solution. Nehru Memorial College (Autonomous), Puthanampatti, Tiruchirapalli, Tamilnadu. India. Test for Steroids: (Liebermann burchard test) The sample was taken in a test tube. Added few drops of glacial acetic acid, Journal of Pharmacy Research Vol.5 Issue 5.May 2012 2895-2899 Meenaa Venkataramani et al. / Journal of Pharmacy Research 2012,5(5),2895-2899 , 18-JAN-2012 + 10:26:22 Vm chloroform extract 18 01 12 Scan EI+ 17.40;191 31.24 TIC 100 21.36;43 24.79;88 57 1.34e9 23.36 32.90 43 57 18.45 29.40 43 57 34.46 22.12 57 % 43 27.67 71 16.28 29.69 43 21.12 25.67 69 5.39 43 91 9.09 14.89 32.20 43 6.15 19.16 22.67 7.74 41 12.01 164 99 43 222 43 43 128 1 Time 5.56 7.56 9.56 11.56 13.56 15.56 17.56 19.56 21.56 23.56 25.56 27.56 29.56 31.56 33.56 Chromatogram (x-axis = Retention time; y-axis = % intensity/% abundance/ concentration) Figure1: Chromatogram representing the peak areas of Phytocomponents matched with the NIST Library acetic anhydride and 1ml of concentrated Sulphuric acid was added along the extract of the plant was performed using PerkinElmer Clarus 500 equipped sides of the test tube. with a column type Capillary Column Elite-5 (5%Phenyl 95% dimethylpolysiloxane), with a column length of 30 m /0.25mm and a film Test for Terpenoids: (Salkowshi test) thickness of 250µm respectively. The extracts were diluted (1µl/ml) and 1µl The substance was warmed with tin and thionyl chloride. was taken as the injection volume. They were injected in the split mode with 10:1 ratio. The oven was programmed at 50ºC @ 8ºC/min to 220ºC (2min) @ Test for Flavonoids: (Shinado’s test) 8ºC/min to 290ºC (10min) and the injector was maintained at 290ºC. Helium To the substance, added alcohol, few magnesium turnings and few drops of was used as a carrier gas with a constant flow at 1 ml/min. The ionization concentrated hydrochloric acid and boiled for 5 minutes. voltage was 70eV. The constituents were identified after comparison with those available in the computer library (NIST) attached to the GC-MS in- Test for Coumarin: strument and the results obtained are reported. A small quantity of substance was mixed with few drops of 10% sodium hydroxide. RESULTS AND DISCUSSION: Preliminary phytochemical screening of various leaf extracts of the plant Test for Quinones: Hiptage benghalensis revealed the presence of Coumarins, Sterols, Lignin, The substance was mixed with few drops of concentrated sulphuric acid. Saponins, Flavanoids, Alkaloids, Tannins, Terpenes and Protein (Table – 1). Several investigations have been attributed to study the phytochemical Test for Lignins: compounds in H.benghalensis and it was found that the results of the present The substance was mixed with alcoholic solution of phloroglucinol and added phytochemical analysis conducted in this study are in accordance to those few drops of concentrated hydrochloric acid. previous reports on the plant (17)(18)(19)(20). Test for alkaloids: Table 1: Phytochemical screening of leaf extracts of Hiptage 1. To the substance few drops of acetic acid was added, followed by benghalensis Dragendroff’s reagent and shaken well. S.NO Test for Chloroform Methanol Ethanol Water 2. The substance was mixed with little amount of dilute hydrochloric acid extract extract extract extract and Mayer’s reagent. 1 Coumarin + - + + Test for Sugars: 2 Sterol + + + - 3 Quinone - - - - The substance was mixed with anthrone and 1 drop of concentrated Sulphuric 4 Lignin + + - + acid and warmed gently. 5 Saponin + + + - 6 Flavonoid - + + + Test for Proteins: 7 Gum - - - - 8 Alkaloid + + + + The substance was mixed with saturated solution of picric acid. 9 Protein + + + + 10 Terpenoid + + + - Gas Chromatography – Mass Spectroscopy analysis 11 Tannin - + + + The Gas chromatography –Mass Spectroscopy analysis of the chloroform + Positive, - Negative. Journal of Pharmacy Research Vol.5 Issue 5.May 2012 2895-2899 Meenaa Venkataramani et al. / Journal of Pharmacy Research 2012,5(5),2895-2899 Table 2: Phytocomponents identified in the Chloroform fraction of Hiptage benghalensis by GC-MS S.No. Peak Name Retention Peak %Peak time area area 1 Name: 5,9-Dodecadien-2-one, 6,10-dimethyl-, (E,E))- 5.39 7370377 0.9609 Formula: C14H24O MW: 208 2. Name: Cyclopentanol, 2-methyl-, acetate, cis- 6.15 3335318 0.4349 Formula: C8H14O2 MW: 142 3. Name: 5-Hepten-2-one, 6-methyl- 7.74 2385087 0.311 Formula: C8H14O MW: 126 4. Name: Pentane, 1-(1-butenyloxy)-, (Z)- 8.49 177112 0.0231 Formula: C9H18O MW: 142 5. Name: D-Limonene 8.7 2713036 0.3537 Formula: C10H16 MW: 136 6.