PJ BS Bossxiczs so Pakistan Journal of Biological Sciences ANSI ez n Network for Scientific Information 308 Lasani Towns Sargodha Road, Faisalabad - PakistanPakistan Joumal of Biological Seienees 10 (11): 194-1946, 2007 ISSN 1028-8880 (© 2007 Asian Network For Selentifie Information. Isolation of Saponin from Dried Roots of Gypsophila simonii Hub. Mor °B, Siti Cevrimli, “Ergin Kariptag and 'A. Nibal Yocekutlu "Department of Chemistry, Faculty of Arts and Seienees, Gazi University, 06500, Ankara, Turkey “Department of Biology, Arts and Sciences Faculty, hi Evran University, 40100 Kargehir, Tarkey Abstract: In tho presont research, a saponin was olaied from the roots of Gypsophila simon. The structure Was charucterized by means of 'H NMR, "C NMR, FTIR and EIMS. The findings indicate that the proposed structure ofthat saponia was as a new Gypsogenin ester (C:, Hy, 0.) Key words: Gypsophia simoni, isolation, saponin stricture INTRODUCTION Saponins (a group of phytoanticipins) are present ‘constitutively in plants and play important roles in plant defense (Figen, 2006), Some saponins of Gspsophila species are broadly used asa drug with entersive medical Importance such as an expectorant and diuretic (Ansari and Has, 1989) and are used in treatments of hepaitis, ‘gestitis and bronchitis Mizaanj¢t al, 1984), A number ‘of saponins were isolated from various Gspsophila species and investigated by many rexearchers (Henry ef al, 1991; Prechet etal, 191; Yay et al, 1998; Acebes etal, 1998; Bemadte and Parante, 2004, Gypsophila simon, local name Céven, isan endemic growing plan at Ganlan province in Turkey (Davis, 1982) ‘There is no study conducted on the eypsogenin ester saponins of this plant to date. In this study, we aim to present the isolation and structural elucidation of new ‘unfamiliar saponin from the root of Gypsophila simon. ‘MATERIALS AND METHODS General: Chemical materials that are ied in the ‘experimen rein analytical grade, Infrared specs of the ‘compounds were recorded between 400 and 400 em” on ‘Matson 1000 FT-IR spectrometers which where clirated tusing polystyrene bands. The reschtion of IR spectrometer is Jom” and the numberof scan is 20. The samples were prepared as aKBr iso “Te meng point was determined in a glass capillary tube, Mass spectrum was recoded on an electron impact, mass spoctrometer from Research Institute of Tubitak (Turkey). HNMR (400 MEIZ, DMSO.) and °C NMR (100 MHZ, DMSO-d.) were explored in Middle East “Teetmical Univesity (METUD. Plant material: Coven (Gypsophila.simonii) was collected around Canin province, Tiskiye, in June 1997 and identified by Professor Zeki Aytag, Department of Biology, Gazi University. The root material was dried in a ‘coal dark place and powdered inthe Faculty of Pharmacy ‘of Gazi University, Isolation and extraction: Collected plant's roots were removed and dried Approximately 35 kg of dried ‘materials were placed into a cartridge and then extracted ‘with chloroform in Soxhlet apparatas for 24h. The ‘cartridge was re-extmcted with ethanol for extra 8 h and ried completely a the room temperature (Baytop, 1991) ‘The extracts containing saponins were evaporated by using a rolary evaporator (Biby, Oklahoma) at 40 rpm, ‘without solvent at the reduced pressure, The dried extracts were dissolved in ethanol and applied on thine layer chromatograms (TLC (20°20, silica gel Gy Art. 7731) in the solvent system (-Bu OH : 1-P1OH: HAc : H,0) (40; 207,5:30), Spots on TLC were detected by spraying with 10% H,S0. followed by heating a 110°C for $-10/min, Spraying was done in order to identify the points of the separated compounds (Segal ef al, 1978). Acid hydrolysis of the saponins: Each of the separated ‘saponins (Gypsogenin ester) was refluxed for 7h with 38% ICI (Ckawa et af, 2002), Third spot (R, = 028) was an up for isolation and identification. Sugar components were identified om Paper Chromatograms, The sugar in filtrate was identified as D-Glueose by comparison on PC (ethyl-acetate; pyridine; ‘water 12, §,4) with on authentic sugar (Boders, 1972). RESULTS AND DISCUSSION ‘The ethanolic extract obtained from the dried roots ‘of Coven (Gypsophila:simonii) was purified on TLC Corresponding Author: ‘Dr. Engin Karipag, Abi Evran Universes, Kuyshir Fen Edeiyat Fables, Biyoloj Bolum, 40100 Kirsew/Taskye Tel: 009038625280S0/134, 0090505290414 Fax: 0903862528054 reatPal. Biol Soi, 10 (1) 1944-1946, 2007 Fig. 1: Gypsogenin ester saponin ‘Tbe LHC ante NOUR dua porn, DMSO-d, “Teble2: FTIR pec dat, corre HBqga — Woremmber en Aion a0 ea 340 i uw C= Ortehingtr ete) 1s618 c= csraching se CHtbentse 1190 cH. yaw tie Ao La fan : 34100 au ow Has0 Loss 3000 : ina ia 1s aS sa so 13500 038 2390 99 34450 Ls exo : 3490 oe nae 55 os Loo (Ry ~ 028). Structure ofthe isolated Gypsogenin ester ‘was characterized by 'H NMR, "C NMR, IR and EIMS, ‘The results were compared with the simular studies (Malaviya ef af, 1991; Zhwo et al, 1990, Delay eal, 1997; Zhao ef al, 1997). Tt was concluded that the structize of Gypsogenin ester may have the fom presented in Fig 1 Data of IR spectrum (KBr, cm) exhibited absorptions at 3400 (OH), 1656-1618 (C = C), 2967-2877 aliphatic C-H stretching and an ester (1714 carbouy stretching -C (0}-OFD, 1190 asymmetric stretching) ‘The proton nuclear magnetic resonance, "Hi NMR. (400 MHZ, DMSO-d, 8 ppm), IR spectral data of, ‘compound (Fig. 1) showed the doubled vinylie proton [6.50-7.00} ‘The carbor-1 3 melear magnetic resonance, °C NMR (400 MHZ, DMSO-d, 8 ppm), spectral of compound (Gig. 1) and its devivatves are shown in Table ypsogenin ester saponin (Fig. 1); mp: 235°C (ancomupted) IR spectrum of studied compound (Fig. 1); ‘and its derivatives are shown in Table 2. Molecular ion peak wan also observed at EEMS: m/z, [M'] 472. All these ‘results confirm that the proposed structuce of that saponin appears oa a new one and is called as Gypsogenin ester (Cs, HO). PC results showed the presence of D-Glucose by ‘comparing their retention times with tote of authentic sugar (R, ~ 1.00, mp= 204°C). 1945,Pal. Biol Soi, 10 (1) 1944-1946, 2007 CONCLUSIONS In recent years, although technology and medicine Ihave developed extensively duc to the decrease in natural richness ond other drawbacks, some oountriss have ‘made it obligatory to use natural products for many goals (Estark etal, 2003). For these reason Gypsophifa species ‘are used forthe treatment of various diseases, ‘The above studies the saponins were extracted from Gypsophiia simonii (Goven) dried roots and then separated its components by thin layer chromatography. NMR, PTIR and EIMS were carried out to investigate ‘unknown saponin presents in this plant. The comparision ‘of this investigation results with similar studies shows that the proposed strucnare of that saponin it neve type and could be named as Gypsogenin ester (Cy, Hy 0,) However litle is known about sugar presence and sigar linkage pattem in saponins of Gypsophila simomi. Paper chremotograms showed that, in Gypsophila. simonii (Ceren), D-Clncose was identified by comparison with on authentic sugar In conclusion, te studies on the saponins highlight the necessity for a comprehensive detailed investigation ‘on the other gyprogenins saponin types which have @ ‘broad medical and industrial potential ACKNOWLEDGMENT We thank kindly to Professor Zeki Aytae in Department of Biology. Gazi University (Tuskiye) for identification of the plant material and moreover, we are grateful to Professor Vilmaz Yildar from Department of| (Chemisty, Gazi University (Tukiye) for help with spectra interpretation REFERENCES ‘Acebes, B, AMA, Az-Lanza and M. Bemabea, 1998. A saponin from the roots of Gypsophila bermejo. Phytochemistry, 49: 2077-2079, Ansari, AK. and 8. Has, 1989, Structural studies on a saponin isolated from the seeds Nigella sativa Phytochemistry, 27: 377-379, Baytop. A. 1991. Farmasstik botani ders kitabs, Yay No. $8, Ist Univ. 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