ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 3297: 2007 Certified Organization) Vol. 2, Issue 12, December 2013, ISOLATION, STRUCTURAL ANALYSIS of an ALKALOID in TODDALIA ASIATICA and THEORETICAL COMPARISON Lawrence T, Gunasckaran S* Research scholar, Department of Physies. Karpagam University, Coimbatore Tamilnadu, India ean, Reseach and Development, Si Pete's Usiversity, Avadi, Chennai Tamilnadu, tadia® Abstract: In the presen study, an attempt has been made to prepare an extract by the methanol from Toddaliaasiaiiea leaf and it contains alkaloids. Tho molecular sructure, funetional groups, chemical bonds, ec has been analyzed with the help of IR, UY and MS spectrum, The Gaussian software has been used to estimate the theoretical values of vibrational frequencies of IR spectrum and for obtaining possible structure ofthe compound. The theoretical values are in good agreemtent with the experimental values and the structure of the alkaloid is confirmed. Keywords: Isolation, IR, UV, MS spectrum analysis, Toddaquinoline alkaloid, LINRODUCTION Nature is the source of medicinal agents for the impressive number of modern drugs for many years. It has beet observed thatthe Toudaliaasiauica, a Todda’s (Tubal) medicinal plant contains Davonoids,steouds, alkaloids atc, [1] and are using by Toddas, a wibal people living in the Nilgiis district, Tamilnadu, India. The extract of Toddalia ‘asiatica plant leaf has been used by them as medicine for fever, wounds, pains etc, [2]. Chen etal. [3] have reported the isolation and characterization of an unusual alkaloid voddaquinolin [4] found in the root bark of Toddalia asic Further David ct al. [5] have symthesized toddaquinoline by the Key step of Cobalt (1) mediated radial cyclisation to a pytidino and achieved, The root bark of Toddalia asiatica a now benza|blquinoline alkaloid synthesized (6]. Toddalia ‘asiatica led tothe islation of 30 compounds, including eoumarins, henzaquinoline, ete 7]. Toddalia asiatica is wsed 15 a folk medicine in Asia relieves pain and stasis [8] “The chemical composition of the Toddafia asiavica has boon Widely studied and identified the toddaquinoline [9]. The aqueous solution of the Toddalia asiatica contains gum and rucilage whereas the methanol extract contains alkaloid, carbohydrates, tannins and falavonoids,etc.[ 10} In the present stidy, an attempt has heen made to compare the experimental values with the theorctial values of vibrational frequencies of IR spectrum of the alkaloid toddaquinoline, evaluated using Gaussian Software and rmoleculr structure by MS spectrum. II, MATERIALS AND METHODS Plant collection: Toddalia arctica is found in the Nilstis istrict, Tamilnadu, India at he aitude around 2200 meter fiom MSL. Smal picks ae all over the stem and botom of leaf. Io this plant, an alkaloid ih in bark and stem [11 ‘Around Ske of halhy. we leaf fee from ust ec. were collected and died ia shalow. Sample Preparation: The dried leves of 1.Skg mechanically crushed and soak in n-Hexane for 72 hours. n-Hexane dissolves wax like moisture compound of eat [12] Then the leaves are filtered from n-Hexane and dred. Again leat soaked in an Acetone for 72 hous, Acetone has more polatity then n-Hexane. Therefore most of the moistues ike ta were dissolves in Acetone. Finally the lea tered and dried completely from aestone then soaked into methanol ina slass container for 72 hours and then the methanol extract alone collected in glass container. The methanol extract prepared fortwo more times from the leat. Methanol exact ken into 5iKiml round boitom flask. A Condenser and mantel arrangement ars used for evaporating the methanol fom extract up to is 1/5 volume and its called methanol concentrate [Fig a]. Again the lest powder sonked in mthanal, the ahove process repeated for 4 times for preparing msthanel cancentat, without drying the lst Copysight wo UIRSET wie crm 1581ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 3297: 2007 Certified Organization) Vol. 2, Issue 12, December 2013 ‘This methanol concentrate contains moisture, lavonoids, steroids ele, Hence it is washed by n-Hexane [Fig bl. The washed methanol concentrate is filler many times by No.l waterman paper for removing the sediments, washed Imojstures and leaf powder. Then itis called filtrate (Fig e]. The filtrate allows drying and adding methanol to it and is called methanol filuate, Fig a. Methanol concentrates preparation, Figb, MeOH Concentrate. Fige, Methanol filtrste Dragendosffs reagent [14] . Dragen dovifs reagent are used for alkaloid test Few drops of Mayss’s seagent added With filuate, while precipitate is obtained. Brown precipitate is oblained afler the addition of few drops of Dragendosffs seagent [15]. Itproves thatthe presence of alkaload Big Column Chromatography Fig Mayer's test(whate precipitate) Fig. TLC test IIL ALKALOID SEPARATION BY COLUMN CHROMATOGRAPHY METHOD. Column chromatography [16] isthe best and quick physical method for separation of individuals from mixtures [Fig d] ‘This is a separation, which works well for ropane alkaloids such as atropine, cocaine, and scopolamine. The metianol extract is fractionated by column chromatography on SiO:, This separates lipids and terpones from the crude alkaloid fraction, The alkaloid traction is chromatographed (SiO: CHC: MeOH = 10:1) to isolate the pure alkaloids. The column was packed by using the suspension of silica gel in n-Hexane. Then the different column elutes were collected 1nd kept in separate containers and are named as SCI SVC2 , SVC3 and S¥C4. Again SUCI, S1/C2, SUC3 and Copyeight to UIRSET. swords com 7592,ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 3297: 2007 Certified Organization) Vol. 2, Issue 12, December 2013 S3C4 ete, under go the alkaloid test with Mayer's regent, SUC3 shows white precipitate, t proves presents of alkaloid [Fig e]. n-Hexane evaporated out and then sample dissolves in pure a-Henane. It is colleted in sample holder and hamed as SUCS, Purlty test. S13 purification is confirmed by Thin Layer Chromatography (TLC) [17] and paper chromatography (PLC). TLC prepared by a sill glass slab coated with Si0> and a small dot of SUC3 Kept just bellow the centre of the lass slab and placed slanting position in 100m glass container with 20ml of n-Hexane, The glass container closed and monitored, After 8 hours the dot moves upward and no further dots of SUIC3 found [Fig]. It shows no other moisture With SUC3. PLC is prepared as a small dot of the SUCS is diffused near the center of small rectangular #1 waterman papor vertically hangover in LODml glass container with 2ml of n-Hexane, IV. EXPERIMENTAL SECTION SHIMADZU spectrometer is used for reconding the IR and UV spectrum. IR spectra of liquid base samples were also recorded by the SHIMADZU. Fourier transforms infrared spectrometer using NaCl cel. EOL GC Mate instrament is used for recorded Mass Spectrum [18-19] Y. RESULT AND DISCUSSION ‘The Gaussian software (3) is used for obtaining IR. UV spectrum and most possible molecular 3D structure of the given TUPAC named compound or molecular diagram. The IUPAC name of Toddaguinoline is obtained by Chemdoo] Software ‘The TUPAC mame ofthe toddaguinoline is 13.15-Dioxa-3-azatetracyelo [8.7.0.0] heptadeca-1(17), 2.4, 6.8, 10, 12(16) -heptaen-5- Theoretical values are compared with experimental result and are listed bellow. Toddaquinoline: TR(cm" Ya, OH-H honded 3$00,3200, N-H Stretch-3381,3356,3931,3315,3298,3284 (secondary amines). CH. Stvetch-2980,2968,2943,2899,2881,2888, 2864, 2839. C-OH Stretch -2750.C=C Stretch-1647, C-C Stretch in ring aromtie-1448,1419. CN Stroteh-1327, 1091, 1055, 1047 C-H loop aromatie-881,802,731 UY 4, 265.5,253 5,239.5 Gaussian software output: RCM puxs O-H Stich 3817, C-H Stretch 3170, 3162, 3193, 3175, 3195,3175, 3216 (C-H Steeteh 4250, CT Stretch 30813033, 3081, 3033, NAC Stretch 1664, L482 , 387. C-C Stretch 1677 1461, 1686 1516,1413, 1686, 1664 ,1550 1387 (1177 1004 .N-C Strteh 1305 ,1225, C-C Stretch 16, 1640,1567, 1664. 1334 1295 , 1413, 1461.1305 .0-C Sueteh 1516,1205,1115, 841, 1072995. 983, C-C-C Bending 855, 610, C-0-C Bending 1072, 983.746, “HOC Bending 1305,1225, 1213, 1177, HCN. Bending 1431,13341L-C.C Bending 1664,1225,1213,1907, H-C-C Bending 1334,1207, 1461,1305,1235,1153, 1516,1251,1213, C-N-C Bending 486, 417, C-C-C Bending 704,506, C-C.N Bening 1004, Bending 0-C-C506, ‘VI.SPECTRAL GRAPHS. IR Spectrum of Toddaquinoline. Copyright o UIRSET swords com 7583ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 3297: 2007 Certified Organization) Vol. 2, Issue 12, December 2013 UV Spectrum of Toddaguinoline. 2 2000 —— a Mass Spectrum Mass Special data, m/e senn,2074, TIC=202512, Base:3.48S, #ion=75, RT=11.93, rn/z, 106.89.128 32,146.50, 161.278.175.367, 191 3012,206.26,241.53,301.68,336,73,340.51, 451.00, 469.77, 468.74 LLM uit Seam, 1S13,TIC=163528, Hase:I.1GES, #ion=63,RT=7.59, nvr -10.85,133.29,170.28, 199.81, 229.28, 276.66,312 95,376.66, Copyright o UIRSET swords com 7584@ ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 3297; 2007 Certtied Organization) Vol. 2, Issue 12, December 2013 rv Sean.711.T1 3392, Base:1 8FS, #ion=15.RT=1.57. vz ~ 69.01,89,87, 105.13, 120.07, 190.27, 151 141,174,125,271.12,281.84,396.01,414.45, Scan. 2372,TIC=280864 BaseS.74FS, #ion-82,RT=12.02, snve:104.33,149,266,177.11,222.16272.74,344.34,374.09 413.52,456.77 LUM T] Copyright o UIRSET swords com 585ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 5297: 2007 Certified Organization) Vol. 2, Issue 12, December 2013 bhad Wurlmntot diay Ae Sean 2383, TIC=269824 Buse 2GFS, Hion=O6.R 11/2269. 126.78.17.90.11,105.049.119.22.132.3. 267-81.281.65.314.45, 343.46. 4.99, 52, 147,56,161.37.176.28.185.39,201.36.22014, 238.33, Wal tr | err Scam 2870, TIC=254448, Base 8%FS, Bion=72, RT=15 43, vp80.054,121 124,134.37, 161 38,202.04,210 51 245.34.999 81,327.54 Copyright o UIRSET swords com 75865@ ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 5297: 2007 Certified Organization) Vol. 2, Issue 12, December 2013 Sean 4116, TIC=170256,Base-44FS, #ion=86,.RT=21 69, “ VFS83.86, 105.017, 122.916, 149.159, 183.57,204 652,247 398,267.616,291.28, 44 56.364 516 4 . ‘TIC min-4.56, 759, 9.07, 11.92, 12.92, 1498, 15.43, 17.17, 21.69, 2398 Copyright o UIRSET swords com 1587ISSN: 2319-8753 IJIRSET International Journal of Innovative Research in Science, Engineering and Technology (An 180 3297: 2007 Certified Organization) Vol. 2, Issue 12, December 2013 VII. concLusion ‘The experimental values of IR, UV and MS are used (o predict the molecular structure, atomic bond, possible molecular functional groups. et. fr the confirmation of toddaquinoline alkaloid present inthe Todd asiatica plant and confirmed. Molecular structure is found by Gaussian software and IUPAC name by the Chenmwioole sofware and reported ss (gj) Fig j, Todaquinoline - Molecular strictures. REFERENCES $ Rajon MSeduraman D Suresh Babur. Anes sines fife. 1(4k360 365.1997 Complencotry and aterstive ratcine 3, Ww [2] Yeeramathe Daripansn, Missin Ayyuna. Satara Laci. HMC Haws, 12.6982.6.35, 2006, [5] Chon. (8, Tel Wa, 4.SboenW SLi TH Poytstem, 311449, 1995, [3] Hamoanen, DC Aan, MAT. Ble Nand Fenwicke DR Tetratciom Lat 8.975, 198 5] DC Harooves, MIT Nu, NJ. Blume sed DR Fenic, Teleated Fever 2001 Jo] Chen TaI Wa, WS SheonT Ishikawa, HL, Poytcbemis Hover, 1953 F7) Hsu MP Wan, CM Teng. 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