Abstract : Medicinal plants are source of important therapeutic aids for alleviating human ailments. Thus natural products have been a major source of drugs for centuries. In tune with this effort, the objective set for the present investigation is to identify the chemical constituents of the leaves and roots ofPremna serratifoliaL. In order to determine the nature of the principle component responsible for its medicinal property. All parts of the plant have medicinal properties. Taking into consideration the medicinal importance of the plant, the volatile organic constituents were analyzed using GCMS (gas chromatographymas spectrometry) and the structures were conformed by genesis. A total 29 compounds from both leaves and roots, where the major compounds are 1HCyclopropeazulen7ol, decahydro1,1,7trimethyl4methylene, 1ar(1aà,4aà,7á,7aá,7bà) (2.98 %), 2Furancarboxaldehyde, 5(hydroxymethyl) (2.44 %), 2Hydroxy3methylbenzaldehyde (6.39 %), :2s,6s2,6,8,8Tetramethyltricyclo5.2.2.0(1,6)undecan2ol (6.35 %), Benzofuran, 2,3dihydro (29.94 %), Glycerin (1.14 %), nHexadecanoic acid (13.94%), 2Propenoic acid, 3(4methoxyphenyl) (13.84 %) have been identified. Key words: Premna serratifoliaL, Volatile Constituents,GCMS.
International Journal of PharmTech Research CODEN (USA): IJPRIF ISSN : 0974-4304 Vol. 3, No.2, pp 1050-1058, April-June 2011 Identification of Volatile Constituents from Premna serratifolia L.through GC-MS C. Ravinder Singh 1 *, R.Nelson 1 , P. Muthu Krishnan 2 and B.Pargavi 3 1*&3 Department of Biotechnology, Vivekanandha college of Arts and Science for women, Thiruchengode – 637 205, Tamilnadu, India. 1 Department of Botany, Govt. Arts College, Ariyalur, Tamil nadu, India. 2 Department of Chemistry, Vivekanandha college of Arts and Science for women, Thiruchengode – 637 205, Tamilnadu, India *Corres.author: chinnaravinder@yahoo.co.in, Phone:09597992543 Abstract : Medicinal plants are source of important therapeutic aids for alleviating human ailments. Thus natural products have been a major source of drugs for centuries. In tune with this effort, the objective set for the present investigation is to identify the chemical constituents of the leaves and roots of Premna serratifolia L. In order to determine the nature of the principle component responsible for its medicinal property. All parts of the plant have medicinal properties. Taking into consideration the medicinal importance of the plant, the volatile organic constituents were analyzed using GC-MS (gas chromatography-mas spectrometry) and the structures were conformed by genesis. A total 29 compounds from both leaves and roots, where the major compounds are 1H- Cycloprop[e]azulen-7-ol, decahydro-1,1,7-trimethyl-4-methylene-, [1ar-(1aà,4aà,7á,7aá,7bà)]- (2.98 %), 2- Furancarboxaldehyde, 5-(hydroxymethyl)- (2.44 %), 2-Hydroxy-3-methylbenzaldehyde (6.39 %), : 2s,6s-2,6,8,8- Tetramethyltricyclo[5.2.2.0(1,6)]undecan-2-ol (6.35 %), Benzofuran, 2,3-dihydro- (29.94 %), Glycerin (1.14 %), n-Hexadecanoic acid (13.94%), 2-Propenoic acid, 3-(4-methoxyphenyl)- (13.84 %) have been identified. Key words: Premna serratifolia L, Volatile Constituents,GC-MS. Introduction Premna serratifolia L. (Verbenaceae) is an important woody, medicinal plant, it is locally known as munnai and has prominent place in Ayurvedha, Siddha, Unani system of medicines 1 . The leaves and roots are astringent, anti-inflammatory, antibacterial properties and are used in cardiac disorder, cough, leprosy, skin disease, constipation, fever, diabetes, obesity, stomach-ache and tumour 2 . And it has cardiotonic 3 , anti- hypoglyceamic properties 4 , anti- coagulant 5 , anti- arthritics 6 and cardio protective effect 7 . The main objective of this study is to identify the chemical constituents from the leaf and root of Prema srratifolia L. This might be responsible for the reported biological activity of this plant. In the present study, Volatile organic matter of the leaves and roots of Premna serratifolia L. was analyzed for the first time. This work will help to identify the new compounds, which may helps to produce important therapeutic products. Material and Methods The plant Premna serratifolia L. was collected from Keelathaniam, Pudukottai District, Tamil Nadu, India and scientific authenticated in Rhbinath herbarium, Trichy, Tamil Nadu, India. Plant Materials Preparation The leaf and root were collected from 8 years old Premna serratifolia L. and washed in tap water C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1051 and then chopped into small fragments. Then materials were dried under shade conditions for 30 days and the drying operation was carried out under controlled conditions to avoid chemical changes. The dried samples were powdered roughly with hands. The powdered samples were stored in polythene containers at room temperature. Extraction of Samples The organic constituents from dried plant tissue (leaf and root) prepared by continuous extracting the powdered materials in Soxhlet apparatus with ethanol as solvent. The extracts were concentrated to one third of their original volume and used for testing the chemical constituents. After completion of extraction, the extract was filtered and concentrated to dryness under hot air oven at 55 o C. The residue appeared as a dark brown powder. GC-MS Programme: Column: Elite-1 (100% Dimethyl poly siloxane), 30m x 0.25mmID x 1µm df Equipment: GC Clarus 500 Perkin Elmer Carrier gas: Helium 1ml/min Detector: Mass detector- Turbo mass gold- Perkin Elmer, Software- Turbomass 5.1. Sample injected: 1µl Split: 10:1 Oven Temperature programme: 110deg-2min hold Up to 280 deg at the rate of 5 deg/ min-9 hold Injector temperature: 250 deg c Total GC time: 45 min MS Programme Library used: NIST Ver.2.1 In let Line temperature: 200 deg c Source temperature: 200deg c Mass scan: (m/z) 45- 450 MS Time: 46 min Phytochemical studies The preliminary phytochemical screening test has been attempted in difference parts of P. serratifolia to find out the presence or absence of certain bioactive compounds Table-1. All the extracts were used to test for the presence of alkaloids, sugar, reducing sugar, catachins, anthroquilnones, amino acids, flavonoids, steroids, terpenoids, tannins, phenolics and saponins. The methods of preliminary phytochemical analysis were based on the methods of Brindha et al 8 1. Prelimary Phytochemical Test (Brindha et al., 1981). Sl.No Test Observation Inference 1. Test solution + minimum quantity of chloroform, 3-4 drops of acetic anhydride and one drop of conc. H 2 SO 4 Purple colour changes to blue or green Presence of steroids 2. Test solution + piece of tin +3 drops of thionyl chloride Violet or purple colour developed Presence of triterpenoids 3. Test solution + 2ml of Fehlings reagent + 3ml of H 2 O Red – orange colour formed Presence of reducing sugars 4. Test solution + very small quantity of anthrone + few drops of conc. H2SO 4 and heat Green to purple colour developed Presence of sugar 5. Test solution taken with 2 NHCL. The aqueous layer formed was decanted and to this one or few drops of Mayers reagent was added White precipitate or turbidity formed Presence of alkaloids 6. Test solution in alcohol + one drop of natural ferric chloride (5%) solution Intense colour developed Presecne of phenolic compounds 7. Test solution in alcohol + Ehrilich reaget and few drops of conc. HCl Pink colour formed Presence of catachins 8 Test solution in alcohol + a bit of magnesium and one or two drops of conc. HCl and heat Red or orange red colour formed Presence of flavanoids 9. Test solution + H 2 O and shake Foamy lather formed Presence of saponins 10 Test solution + H 2 O + lead acetate White precipitate formed Presence of Tannins 11. Test solution + magnesium acetate solution Pink colour formed Presence of anthroquiones 12. Test solution + 1 % ninhydrin in alcohol Blue or violet colour developed Presence of amino acids C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1052 Table-2 Phytochemicals identified in the alcoholic extract of the leaf Premna serratifolia Fig-1 Phytochemicals identified in the alcoholic extract of the leaf Premna serratifolia Sl. No RT Name of the compound Formula MW Peak area % 1 4.09 : Glycerin C 3 H 8 O 3 92 2.79 2 5.80 2,5-Furandione, 3-methyl- C 5 H 4 O 3 112 9.27 3 9.20 Benzofuran, 2,3-dihydro- C 8 H 8 O 120 29.94 4 14.74 2-Hydroxy-3-methylbenzaldehyde C 8 H 8 O 2 136 6.39 5 17.43 Dodecanoic acid C 12 H 24 O 2 200 7.88 6 20.54 2-Propenoic acid, 3-(4-methoxyphenyl)- C 10 H 10 O 3 178 13.84 7 21.18 Phenol, 4-(3-hydroxy-1-propenyl)-2- methoxy- C 10 H 12 O 3 180 1.54 8 21.49 2-Propenoic acid, 3-(4-methoxyphenyl)-, ethyl ester C 12 H 14 O 3 206 1.35 9 23.73 1,2-Benzenedicarboxylic acid, bis(2- methylpropyl) ester C 16 H 22 O 4 278 2.50 10 25.84 n-Hexadecanoic acid C 16 H 32 O 2 256 13.94 11 28.71 Phytol C 20 H 40 O 296 6.78 12 29.96 Octadecanoic acid, ethyl ester C 20 H 40 O 2 312 1.68 13 35.62 Octasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13,15,15- hexadecamethyl- C 16 H 50 O 7 Si 8 578 2.11 PPRC TANJORE, 25-Jul-2007 + 15:39:07 Premna serratifolia-Leaf A.lcoholic ext 4.76 9.76 14.76 19.76 24.76 29.76 34.76 39.76 44.76 T i me 0 100 % medicinal plant analysis139 Scan EI+ TIC 8.41e7 9.20 5.79 4.09 5.97 7.79 25.84 20.54 17.4314.74 11.49 19.15 23.73 21.18 40.99 29.51 28.71 38.07 35.62 33.30 29.96 C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1053 Table-3 Phytochemicals identified in the alcoholic extract of the root Premna serratifolia Fig-2 Phytochemicals identified in the alcoholic extract of the root Premna serratifolia Sl. No RT Name of the compound Molecular Formula MW Peak Area% 1 4.08 : Glycerin C 3 H 8 O 3 92 1.14 2 5.79 2,5-Furandione, 3-methyl- C 5 H 4 O 3 112 2.89 3 9.06 2-Furancarboxaldehyde, 5-(hydroxymethyl)- C 6 H 6 O 3 126 2.44 4 9.20 Benzofuran, 2,3-dihydro- C 8 H 8 O 120 9.86 5 14.74 2-Hydroxy-3-methylbenzaldehyde C 8 H 8 O 2 136 34.58 6 16.24 Seychellene C 15 H 24 204 2.30 7 17.43 Dodecanoic acid C 12 H 24 O 2 200 0.71 8 18.30 1H-Cycloprop[e]azulen-7-ol, decahydro-1,1,7- trimethyl-4-methylene-, [1ar- (1aà,4aà,7á,7aá,7bà)]- C 15 H 24 O : 220 2.98 9 20.65 2-Propenoic acid, 3-(4-methoxyphenyl)- C 10 H 10 O 3 178 13.99 10 22.71 2s,6s-2,6,8,8- Tetramethyltricyclo[5.2.2.0(1,6)]undecan-2-ol C 15 H 26 O 222 6.35 11 23.56 3,7,11,15-Tetramethyl-2-hexadecen-1-ol C 20 H 40 O 296 1.34 12 25.85 n-Hexadecanoic acid C 16 H 32 O 2 256 4.87 13 28.71 Phytol C 20 H 40 O 296 1.90 14 29.96 Octadecanoic acid, ethyl ester C 20 H 40 O 2 312 0.59 15 32.69 2-Phenanthrenol, 4b,5,6,7,8,8a,9,10-octahydro- 4b,8,8-trimethyl-1-(1-methylethyl)-, (4bS-trans)- C 20 H 30 O 286 4.77 16 37.01 unknown *** *** 9.29 PPRC TANJORE, 25-Jul-2007 + 16:35:43 Premna serratifolia-Root A.lcoholic ext 4.34 9.34 14.34 19.34 24.34 29.34 34.34 39.34 44.34 T i me 0 100 % medicinal plant analysis140 Scan EI+ TIC 1.61e8 37.01 20. 66 9.20 5.78 4.07 7.79 18. 29 16. 24 11. 89 11.51 14.74 13.48 17. 43 19. 51 22. 71 32. 69 25. 85 23.56 29. 52 28.71 32.57 35. 34 37.40 40. 99 C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1054 (replib) 1H-Cycloprop[e]azulen-7-ol, dec ahyd ro-1,1,7-trimethyl-4-methylene-, [1ar-(1aà,4aà,7á,7aá,7bà)]- 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 0 50 100 41 43 51 53 55 59 65 67 71 73 77 79 81 85 91 93 95 97 105 119 131 145 159 162 173 177 187 202 205 220 OH Fig-3 Name: 1H-Cycloprop[e]azulen-7-ol, decahydro-1,1,7-trimethyl-4-methylene-, [1ar-(1aà,4aà,7á,7aá,7bà)]- Formula: C 15 H 24 O MW: 220 CAS#: 6750-60-3 NIST#: 107043 ID#: 1780 DB: replib Other DBs: None Contributor: N.W. Davies, Centr. Sci. Lab., Univ. Tasmania, Hobart, Australia 10 largest peaks: 43 999 | 41 709 | 91 647 | 105 555 | 119 524 | 93 449 | 205 440 | 79 379 | 159 365 | 55 343 | (replib) 2-Furanc arboxaldehyde, 5-(hydroxymethyl)- 40 50 60 70 80 90 100 110 120 130 140 0 50 100 41 42 44 49 51 52 53 55 66 68 69 81 95 97 98 109 123 126 OH O O Fig-4 Name: 2-Furancarboxaldehyde, 5-(hydroxymethyl)- Formula: C 6 H 6 O 3 MW: 126 CAS#: 67-47-0 NIST#: 60544 ID#: 12832 DB: replib Other DBs: None Contributor: D.HENNEBERG, MAX-PLANCK INSTITUTE, MULHEIM, WEST GERMANY 10 largest peaks: 97 999 | 126 710 | 41 668 | 39 336 | 69 289 | 29 173 | 125 136 | 109 131 | 53 131 | 51 100 | C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1055 (ma inlib ) 2-Hyd roxy-3-methylb enza ldehyd e 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 0 50 100 51 55 59 63 65 73 75 77 79 86 90 105 107 118 133 136 O OH Fig-5 Name: 2-Hydroxy-3-methylbenzaldehyde Formula: C 8 H 8 O 2 MW: 136 CAS#: 824-42-0 NIST#: 72528 ID#: 80172 DB: mainlib Other DBs: None Contributor: R.SELF, AGRIC. RES. COUNC., FOOD RES. INST., NORWICH, U. 10 largest peaks: 136 999 | 135 977 | 77 414 | 79 321 | 90 316 | 107 242 | 51 175 | 53 147 | 73 140 | 118 139 | ( m a i n lib ) 2s,6s-2,6,8,8-Te t ra m eth y ltric y c l o [5.2.2.0( 1,6) ] un d e c a n -2-o l 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 0 50 100 41 43 47 53 55 57 65 67 69 71 77 79 81 83 85 87 91 93 95 109 113 121 135 149 161 175 179 189 204 207 222 OH Fig-6 Name: 2s,6s-2,6,8,8-Tetramethyltricyclo[5.2.2.0(1,6)]undecan-2-ol Formula: C 15 H 26 O MW: 222 CAS#: N/A NIST#: 140230 ID#: 9434 DB: mainlib Other DBs: None Contributor: B. Derendyaev, Novosibirsk Institute of Organic Chemistry 10 largest peaks: 43 999 | 109 928 | 83 469 | 69 438 | 161 425 | 222 421 | 164 404 | 81 392 | 41 386 | 95 379 | C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1056 (replib) Benzofuran, 2,3-d ihydro- 40 50 60 70 80 90 100 110 120 130 140 0 50 100 40 45 51 52 55 60 62 63 64 65 75 77 89 90 91 92 102 105 120 O Fig-7 Name: Benzofuran, 2,3-dihydro- Formula: C 8 H 8 O MW: 120 CAS#: 496-16-2 NIST#: 109771 ID#: 16183 DB: replib Other DBs: None Contributor: Philip Morris R&D 10 largest peaks: 120 999 | 91 855 | 119 288 | 92 216 | 39 159 | 63 134 | 65 122 | 89 121 | 12196 | 5180 | (m a in lib ) Glyc erin 10 20 30 40 50 60 70 80 90 100 0 50 100 14 15 19 27 29 30 31 32 41 42 43 45 55 58 61 62 74 OH OH HO Fig-8 Name: Glycerin Formula: C 3 H 8 O 3 MW: 92 CAS#: 56-81-5 NIST#: 229308 ID#: 6829 DB: replib Other DBs: None Contributor: Japan AIST/NIMC Database- Spectrum MS-NW-5532 10 largest peaks: 61 999 | 43 740 | 44 462 | 31 289 | 15 273 | 29 151 | 60 90 | 18 68 | 27 61 | 28 58 | C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1057 (replib) n-Hexadecanoic acid 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 0 50 100 55 60 69 73 83 97 101 115 125 129 143 157 171 185 199 213 227 239 256 OH O Fig-9 Name: n-Hexadecanoic acid Formula: C 16 H 32 O 2 MW: 256 CAS#: 57-10-3 NIST#: 251929 ID#: 1725 DB: replib Other DBs: None Contributor: Div. of Experiment Therapeutics WRAIR, WRAMC, Washington DC 20307 10 largest peaks: 43 999 | 41 753 | 60 580 | 55 526 | 73 523 | 57 476 | 69 236 | 71 193 | 45 132 | 42 119 | (replib) 2-Propenoic a cid, 3-(4-methoxyphenyl)- 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 0 50 100 41 45 51 53 63 69 75 77 79 87 89 105 107 118 133 135 137 145 147 161 163 178 OH O O Fig-10 Name: 2-Propenoic acid, 3-(4-methoxyphenyl)- Formula: C 10 H 10 O 3 MW: 178 CAS#: 830-09-1 NIST#: 75919 ID#: 22426 DB: replib Other DBs: None Contributor: RADIAN CORP 10 largest peaks: 178 999 | 161 361 | 177 286 | 77 246 | 89 233 | 133 232 63 194 | 132 159 | 79 154 | 90 127 | C. Ravinder Singh et al /Int.J. PharmTech Res.2011,3(2) 1058 Result and Discussion Volatile organic compounds are products of plant secondary metabolites, consisting of complex mixture of mono-, di-, tri- terpene hydrocarbons and oxygenated biological materials. In the present investigation, 13 compounds (Fig-1) from leaves and 16 compounds (Fig-2) from root of Premna serratifolia L. were identified through GC-MS analysis (Table-2,3). Among the total compounds, 8 major compounds A total 29 compounds from both leaves and roots, where the major compounds are 1H- Cycloprop[e]azulen-7-ol, decahydro-1,1,7-trimethyl-4- methylene-, [1ar-(1aà,4aà,7á,7aá,7bà)]- (2.98 %) Fig- 3, 2-Furancarboxaldehyde, 5-(hydroxymethyl)- (2.44 %) Fig-4, 2-Hydroxy-3-methylbenzaldehyde (6.39 %) Fig-5, : 2s,6s-2,6,8,8-Tetramethyltricyclo [5.2.2.0 (1,6)]undecan-2-ol (6.35 %) Fig-6, Benzofuran, 2,3- dihydro- (29.94 %) Fig-7, Glycerin (1.14 %) Fig-8, n- Hexadecanoic acid (13.94%) Fig-9, 2-Propenoic acid, 3-(4-methoxyphenyl)- (13.84 %) Fig-10 have been identified. from both leaves and roots have been identified with comparisons of mass spectrum (Fig-3). The similar findings (Volatile organic compounds) reported from Mimusops elengi 9 , Acorus calamus, 10 , Morinda morindoides 11 . Our study suggests that Premna serratifolia L. may be a potential source of the reported important disease use in medicine. Acknowledgement Author acknowledgement the valuable help rendered by Dr Jhon Britto, Director, Rhabinath herbarium, Thiruchirapalli, Tamil nadu, India for identification of plant specimen. And Mr S.Kumaravel , Scientist, Food analysis Laboratory, Paddy Processing Research Centre (Ministry of Food Processing Industries, Government of India), Thanjavur-613 005. References 1. Ajitkar, B.K., Choudry. and Bandyopadhyay, N.G., Comparative evaluation of hypoglyceamic activity of some Indian medicinal plants in Alloxan diabetic rats, Journal of ethanopharmacology, 2003, 84,105-108. 2. Prajapati, N.D, Purohit, S.S., Sharma, A.K. and Kumar, T., hand book of medicinal plants: A complete source book. Agrobios, 2006, (India). 3. Rekha, R., Susela, I., Meenakshi sundaram, R. and Saleem Basha, N., Cardiostimulant activity of bark and wood of Premna serratifolia L.,Bangladesh Journal of Pharmacological society, 2008, 3, 107-113. 4. Dash, G.K., Patrol, C.P. and Maiti, A.K., A study on the anti-hypoglyceamic effect of roots of Premna corymbosa Rotl., Journal of Natural Remedies, 2005, 5(1),31-34. 5. Gopal, R.H. and Purushothaman, K.K., Effect of plant isolates on coagulation of blood: An in- vitro study, Bull Med. Ethanobot Res, 1984, 5, 171-177. 6. Rathore, R.S., Prakash, A. and Singh, P.P., Premna integrifolia L. Apreliminary study of anti-inflammatory and anti-arthritic activity, Rheurnatism, 1977, 12,130-134. 7. Rekha, R. and Saleem Basha, N., Cordio prective effect of ethanol extracts of stem-bark and stem wood of Premna serratifolia (Verbenaceae), Research Journal of Pharmacy and Technology, 2008, 1(4), 487-491. 8. Brindha, P., Sasikala, K. and Purushoth, K., Preliminary Phytochemical studies in higher plants. Ethnobotany, 1977, 3, 84-96. 9. Anjali, R., Rasika, T., Amruta, T., Vedavati, P. and Nirmala, D., GC-MS study of a stem volatile matter from Mimusops elengi, International journal of chem.Tech. research, 2009, 1(2), 158-161. 10. Rajendra, G. and Kyoung-su Kim., Volatile organic compounds of medicinal values from Napalese Acorus calamus L., Kathmandu University journal of science engineering and Technology,2009, 5, 51-65. 11. Bi Kouame, F.P., Bedi, G., Koffi, A.M., Chalchat, J.C. and Guessan, T.Y., Volatiles constituents from leaves of Morinda morindoides (Rubiaceae): A medicinal plant from the Ivory Coast, The open Natural Products Journal, 2010, 3,6-9. ***** . 1050-1058, April-June 2011 Identification of Volatile Constituents from Premna serratifolia L.through GC-MS C. Ravinder Singh 1 *, R.Nelson 1 , P. Muthu Krishnan 2 and B.Pargavi 3 1*&3 Department. consideration the medicinal importance of the plant, the volatile organic constituents were analyzed using GC-MS (gas chromatography-mas spectrometry) and the structures were conformed by genesis. A total. 3-(4-methoxyphenyl)- (13.84 %) have been identified. Key words: Premna serratifolia L, Volatile Constituents ,GC-MS. Introduction Premna serratifolia L. (Verbenaceae) is an important woody, medicinal plant,