Varadarassou Mouttaya Mounnissamy et al. / Journal of Pharmacy Research 2011,4(1),237-240 Research Article Available online through ISSN: 0974-6943 http://jprsolutions.info Phytochemical investigation of Cansjera rheedii J.Gmelin (Opiliaceae) Varadarassou Mouttaya Mounnissamy1*, Subramanian Kavimani1, Sabarimuthu Darlin Quine2, Kuppuswamy Subramani3 1College of Pharmacy, Mother Theresa Post Graduate and Research Institute of Health Sciences (MTPG & RIHS), (A Govt. of Puducherry Institution), Indira Nagar, Gorimedu, Puducherry-605 006, India. 2School of Chemical and Biotechnology, SASTRA University, Thirumalaisamudhram, Thanjavour-613 412, Tamil Nadu, India. 3Department of Chemistry, Kanchi Mamunivar Centre for Post-Graduate Studies (KMCPGS), Lawspet, Puducherry-605 008, India. Received on: 15-09-2010; Revised on: 15-10-2010; Accepted on:13-12-2010 ABSTRACT A Phytochemical study of an ethanol extract from the aerial parts of Cansjera rheedii J.Gmelin(Opiliaceae), a traditional Indian medicinal plant, led to the isolation of 5 compounds, including a mixture of 2 phenyl propenoic acid derivatives and 3 flavonols glycosides: 3, 4-dihydroxy cinnamic acid (1), 4-hydroxy 3-methoxy cinnamic acid (2), 3, 5, 7, 3’, 4’-pentahydroxy flavone (quercetin) (3), 5, 7, 3’, 4’-tetrahydroxy -3-O-ß-D-glucopyranosyl flavones (quercetin-3-O-ß-glucoside) (4) and 5, 7, 3’, 4’-tetrahydroxy-3-O-(6-O-a-L-rhamnopyanosyl)-ß-D-glucopyranosyl flavone (quercetin-3-O-ß-rutinoside) (5). Structures were established by spectroscopic and chemical method. This is the first time that 5 compounds have been isolated and identified from this plant. Key words: Cansjera rheedii, Opiliaceae, Flavonoids, quercetin, quercetin-3-O-ß-D- glucoside, quercetin-3-O-ß-D-rutinoside. INTRODUCTION Cansjera rheedii J Gmelin (Opiliaceae) is a climbing shrub, sometimes armed, Extraction and isolation generally found in India through Malaya to Hong Kong and North Australia [1-2]. The air dried and coarsely powdered aerial parts (1.0Kg) were extracted with The tribes of Nilgiris in Tamil Nadu, India using the plant extract for the treat- boiling 95% ethanol (3 X 5l) and the extract was concentrated to about 250 ml. ment of post-natal pain [3], intermittent fever [4] and poisonous bites and skin The insoluble green residue was removed by filtration and the soluble in the filtrate diseases [5]. In our earlier studies, the ethanol extract of aerial parts of C.rheedii (150 ml) were fractioned into C6H6, Et2O, EtOAc and EtCOMe. The C6H6 frac- has been reported to have hepatoprotective [6], cytotoxic [7], anthelmintic [8], tion after concentration yielded a pale yellow needle, recrystallized from MeOH anti-inflammatory and membrane stabilizing property [9], antipyretic [10], anti- and designated as compound 1(910mg). The Et2O concentrate was column nociceptive [11] and diuretic [12] activities. The preliminary phytochemical chromatographed over sephadex LH-20 using MeOH. 35 fractions of 50ml each screening reveals the presence of flavonoids, Phenolic and aromatic acids [13]. In were collected. Fraction 4-29 gave colourless needles, recrystallized from MeOH the absence of any report on a systematic examination of this plant and in and designated as compound 2(800mg). The EtOAc concentrate was column continuation of our studies on the flavonoids of Indian medicinal plants [14-15], chromatographed over Sephadex LH-20 using MeOH. 44 fractions of 50ml each the aerial parts of C.rheedii were investigated for polyphenolics and the results where collected, fractions 7-32 gave yellow needles, recrystallized with MeOH and leading to the isolation and characterization of 3, 4-dihydroxy cinnamic acid (1), designated as Compound-3(1.1g). The EtCOMe concentrate was chromatographed 4-hydroxy 3-methoxy cinnamic acid (2), 3, 5, 7, 3’, 4’-pentahydroxy flavone on a column of Sephadex LH-20 using MeOH as eluent. 107 fractions of 50ml (quercetin) (3) 5, 7, 3’, 4’-tetrahydroxy -3-O-ß-D-glucopyranosyl flavones (quer- each were collected, fractions 6-35 deposited a homogenous yellow solid recrys- cetin-3-O-ß-D-glucoside) (4) and 5, 7, 3’, 4’-tetrahydroxy-3-O-(6-O-a-L- tallized from MeOH and designated as compound-4 (89mg). Fractions 36-98 gave rhamnopyanosyl)-ß-D-glucopyranosyl flavone (quercetin-3-O-ß-D-rutinoside) (5) a pale yellow homogenous solid, recrystallized from MeOH and were designated as are here presented. This is the first time that these compounds are reported from compound-5(530mg). the plant. 3, 4-dihydroxy cinnamic acid (1) EXPERIMENTAL Pale yellow needles; mp. 210.5°C gave brisk effervescence with saturated NaHCO3 General experimental procedures 3+ solution [16], light blue with Fe and decolourised Br2 water. Blue under UV and 1D and 2D NMR spectra were recorded on a JEOL 600 MHZ spectrometer, che- deep blue under UV/NH . UV l (MeOH): 240, 324 nm; (+ NaOMe) 250, 302sh, mical shifts (ppm) are related to (CH ) Si as TMS as internal standard. Optical 3 max 3 4 343 nm; (+ CH3COONa) 244, 295sh, 327 nm; (+CH3COONa /H3BO4) 244, 298sh, rotations were determined on a JASCO P-1020 Polarimeter in MeOH. Elemental 326 nm; (+AlCl ) 246, 305sh, 341nm; (+AlCl /HCl) 244, 300sh, 328 nm; 1H analysis by CHNSO Analyser (Thermofinnigan -Flash EA 1112 series). IR spectra 3 3 NMR (500MHz), DMSO-d6: d 7.38 (d, J=16.0 Hz, 1H, H-a); d 6.99 (d, J=2.3 Hz, were recorded on a Perkin-Elmer FTIR spectrometer. UV-Visible spectropho- 1H, H-2); d 6.92 (dd, J=8.4 & 2.3 Hz, 1H, H-6); d 6.72 (d, J=8.4 Hz, 1H, H-5); d tometer (Shimadzu-UV-2500PC series) of each compound was determined in MeOH 13 6.15 (d, J=16.05 Hz, 1H, H-ß). C NMR (500MHz), DMSO-d6: d 168.51(s, and after addition of different shift reagents such as AlCl3, AlCl3/HCl, CH3COONa, =C=O); d148.64 (s, C-ß); d146.05 (C-3); d145.16 (C-4); d126.19 (C-1); d 121.79 CH3COONa/H3 BO4 and NaOMe at 190-500nm. Mass spectra were recorded on (C-6); d 116.25 (C-5); d115.59 (C-2); dd 115.0 (C-a). MS (-ve) observed m/z: 179 GCMS-Celuras-500 (Perkin-Elmer). Melting point determinations by Differen- + (M , 100) calculated for C9H8O4; 180 (M+H); 178 (M-H); IR (gmax, KBr): 3427, tial Scanning Calorimeter (DSC-60) (Shimadzu Co., Japan). Open column chro- 2558, 1654, 1605, 1534, 1449, 1283, 1217, 1176, 1110, 969, 901, 851, 809 matography was carried out on Sephadex LH-20 (Amersham Pharmacia Biotech cm-1. Elemental analysis (%): C (60.69): H (4.24): O (0.33). Co., UK) as packing material and Whatmann No.1 filter paper and TLC-Silica gel 60 F254 sheets( Merck Co., Germany) 4-hydroxy 3-methoxy cinnamic acid (2) Plant material Colourless needles, mp. 210.8°C gave effervescence with NaHCO3 solution, 3+ The aerial parts of the plant C.rheedii (Opiliaceae) were collected from Auroville, decolourized Br2 water and green colour with Fe [16]. It was blue under UV Puducherry in June 2006 and it was identified and authenticated by Auro Her- changing to bright blue under UV/NH3; UV lmax(MeOH): 233, 296, 319nm; barium Sakthi Botanical Survey Department, Auroville, India . A Voucher speci- (+NaOMe): 234, 304sh, 346; (+ CH3COONa): 227, 284sh, 323nm; (+CH3COONa / H BO ): 224, 296sh, 322nm; (+AlCl ):237, 302sh, 331nm; (+AlCl /HCl): 234, men has been kept in our laboratory for future reference (VS-12). 3 4 1 3 3 297sh, 323nm; H NMR (500MHz), DMSO-d6; d 7.46 (d, J=16.0 Hz, 1H, H-a); *Corresponding author. d 7.24 (d, J=1.55 Hz, 1H, H-2); d 7.05 (dd, J=1.55 & 1.50 Hz, 1H, H-6); d 6.76 (d, 13 Research Schloar, J=8.4 Hz, 1H, H-5); d 6.34 (d, J=16.05 Hz, 1H, H-ß). C NMR (500MHz), SASTRA University, Thanjavour, T.N. DMSO-d6; d 168.55 (C-9); d 148.42 (C-7); d 149.59 (C-3); d145.05 (C-4); d Tel.: +91-9442288084 126.28 (C-1); d 123.37 (C-6); d 116.12 (C-5); d111.60 (C-2); d116.01 (C-8); d E-mail: [email protected] Journal of Pharmacy Research Vol.4.Issue 1. January 2010 237-240 Varadarassou Mouttaya Mounnissamy et al. / Journal of Pharmacy Research 2011,4(1),237-240 56.18 (C-10). MS(-ve): (m/z, rel. int. %) 193(M+, 100%) calculated for C10H10O4; Figure 1. 3, 4-dihydroxy cinnamic acid (Compound: 1) 194(M+H); 192 (M-H); IR (gmax, cm-1, KBr): 3436, 2903, 2841, 1686, 1609, 1514, 1424, 1277, 1173, 1116, 1033, 941, 852, 803, 749. Elemental analysis O (%): C (61.53): H (5.20): O (33.27) OH 3, 5, 7, 3’, 4’-pentahydroxy flavone : quercetin (3) HO Yellow needles, mp.305.83°C, gave yellow colour with NH3, Na2CO3 and NaOH, pink with Mg-HCl and olive green with Fe3+[16]. It was yellow under UV and UV/ OH NH3: UV lmax (MeOH): 256, 271sh, 305sh, 373nm; (+NaoMe) 243sh, 330, 432 nm(decompose); (+CH3COONa) 266, 305sh, 358sh, 427nm; (+CH3COONa / H3BO4) 265, 299sh, 362sh, 426nm; (+AlCl3 ) 270, 302sh, 332sh, 447nm; (+AlCl3/ Figure 2 . 4-hydroxy-3-methoxy cinnamic acid (Compound: 2) 1 HCl) 265, 302sh, 359sh, 426nm; H NMR (500MHz), DMSO-d6; d 12.45 (s, 1H, 5-OH); d10.75 (s, 1H, OH-7); d 9.37 (s, 1H, OH-3); d 9.29 (s, 1H, OH-3’); d 9.57 HO (s, 1H, OH-4’); d 7.65 (d, J= 2.3 Hz, 1H, H-2’); d 7.50 (dd, J=8.3 & 2.3Hz, 1H, H- 6’); d 6.87 (d, J=3.8Hz, 1H, H-5’); d 6.38 (d, J=5..3Hz, 1H, H-8); d 6.16 (d, J=1.55 13 OH Hz, 1H, H-6).