THE FT-IR SPECTROMETRIC STUDIES OF VIBRATIONAL BANDS OF SEMECARPUS ANACARDIUM LINN.F. LEAF, STEM POWDER AND EXTRACTS

Academic Sciences Asian Journal of Pharmaceutical and Clinical Research Vol 6, Suppl, 3 ISSN - 0974-244 Research Article ABSTRACT THE FT-IR SPECTROMETRIC STUDIES OF VIBRATIONAL BANDS OF SEMECARPUS ANACARDIUM LINN.F. LEAF, STEM POWDER AND EXTRACTS PARAG A. PEDNEKAR, BHANU RAMAN Department of Chemistry, K.J. Somaiya College of Science and Commerce, Vidyavihar, Mumbai 0 077, Email: professor.bhanuraman@gmail.com Received:0 December 2, Revised and Accepted:2 january 3 The present study was aimed to identify the functional groups present in the crude powder and extracts of Semecarpus anacardium Linn.f. leaves and stem through FT-IR spectrometry. 4 different polar and nonpolar organic solvents were used in the soxhlet extraction of the plant material. All the vibrational bands originated from different groups like C-H, C=C, C C, C-O, C=O, C-I, C-Cl, C-Br, C-F, >CH2, C-N, C=C-C, S-S, N-H, NO2, H-X, O-H, P-O- C etc., and may be termed as key bands and can be studied for their absorption characteristics to differentiate individual plant parts like leaves, stem etc. The FT-IR analysis of leaf powder of Semecarpus anacardium proved the presence of alcohols, phenols, alkanes, alkenes, carboxylic acids, ethers, esters, aliphatic iodo and polysulfides. The FT-IR analysis results of stem powder of Semecarpus anacardium revealed the presence of alcohols, phenols, alkanes, alkenes, amines, amides, aldehydes, ketones, esters, ethers carboxylic acids, nitro, aliphatic iodo. The FT-IR studies revealed the different characteristic peak values with various functional groups. The results of the present study generated the FT-IR spectrum profile for the medicinally important plant Semecarpus anacardium and can be used to identify the plant in the pharmaceutical industry. Keywords:Semecarpus anacardium Linn.f., FT-IR spectrometry, functional groups. INTRODUCTION Fourier transform infrared spectrometry (FT-IR) is one of the most widely used methods to identify the chemical constituents and elucidate the structures of and has been used as a requisite method to identify medicines in Pharmacopoeia of many countries. FT-IR has played a vital role in pharmaceutical analysis in recent years. FT-IR spectrometry is a physico-chemical analytical technique that does not determine concentrations of individual metabolites but provides a snapshot of the metabolic composition of a tissue at a given time 2. The FT-IR method measures predominantly the vibrations of bonds within chemical functional groups and generates a spectrum that can be regarded as a biochemical or metabolic fingerprint of the sample 3. Infrared spectroscopy provides a useful method for herbal analysis as well as for quantitative analysis of drugs and with the help of FT-IR continuous monitoring of the spectral baseline and simultaneous analysis of different components of the same sample is possible 4-6. Applying metabolomic techniques to pharmacognosy as a marker is a new approach, generally used to identify the functional groups 7. Semecarpus anacardium Linn.f. (Family- Anacardiaceae) commonly known as Bhallataka or marking nut is used in indigenous systems of medicine for the treatment of various diseases 8,9. It is a deciduous tree distributed in the sub-himalayan tract and in tropical parts of India. Many mainly biflavonoids, phenolics, bhilawanols, sterols, anacardic acid and glycosides have been identified as constituents of Semecarpus anacardium nut extract. On the basis of chemical and spectral data, several biflavonoids such as jeediflavanone, galluflavanone, nallaflavanone, semecarpetin and anacarduflavanone have been characterized 0. Several monophenolic known as semecarpol and bhilawanol were also isolated. Further analysis revealed the presence of iron, copper, sodium, calcium and aluminium in traces 2 and also the plant is good source of minerals like sulphur, magnesium, phosphorus 3. The present study was intended to resolve the functional constituents present in the leaves and stem of Semecarpus anacardium which will be useful for the proper identification of the active and the chemical profiles will be used as a pharmacognostic marker to differentiate the adulterant from the commercial samples. MATERIALS AND METHODS Plant collection and authentification The leaves and stem of Semecarpus anacardium were collected from an open field around Mumbai, Maharashtra. The identification of the plant was done at the Blatter Herbarium, St. Xavier s College, Mumbai. The plant specimen matches with the Blatter Herbarium specimen no.t-472 of S. C. Tavakari. Leaves were shade dried and made into coarse powder with mechanical grinder and then passed through sieve, B.S.S Mesh No.. Apparatus Bruker Alpha-T FTIR equipped with universal sampling module, equipped with a room temperature DTGS detector. The scan range was taken from 0 to 00 cm with a resolution of 4 cm. Chemicals Chloroform, Toluene, Carbon tetrachloride, Ethyl acetate, Hexane, Ethyl alcohol, Methanol, Acetone, 2-propanol, Petroleum ether (- 80 C), 2-butanone, Dichloromethane, Ethyl ether were all A.R. grade (Merck, India), Distilled water. Preparation of the plant extract The leaf powder of Semecarpus anacardium ( gms) was extracted with 0 ml each of polar and nonpolar solvents by soxhlet extraction for 8 hrs. The extracts obtained were later kept for evaporation to remove the excess solvents. These extracts were then stored in plastic bottles in refrigerator for FT-IR analysis. Powdered leaf material was extracted using water, chloroform, toluene, carbon tetrachloride, ethyl acetate, hexane, ethyl alcohol, methanol, acetone, 2-propanol, petroleum ether (-80 C), 2-butanone, dichloromethane, ethyl ether. The crude powder and dried extracts were mixed with KBr salt using mortar and pestle and compressed into a thin pellet. RESULTS AND DISCUSSION The FT-IR spectrum was used to identify the functional groups of the active components based on the peak value in the region of infrared radiation. The leaf and stem powder and extracts of Semecarpus anacardium were analysed by the FT-IR spectrometers and the functional groups of the components were separated based on its peak ratio. The FT-IR analysis of leaf powder and different leaf extracts of Semecarpus anacardium proved the presence of alcohols, phenols, alkanes, alkenes, carboxylic acids, ethers, esters, polysulfides and aliphatic iodo etc., [Table -, Fig - ]. The FT-IR analysis results of stem powder and stem extracts of Semecarpus anacardium revealed the presence of alcohols, phenols,

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 alkanes, alkenes, amines, amides, aldehydes, ketones, esters, ethers [Table 6-8, Fig 6-8]. carboxylic acids, nitro, aliphatic iodo etc., Table : FT-IR Peak Values of Semecarpus anacardium Leaf Powder Functional 33 O-H Hydrogen bonded 30-30 299.69 C-H Alkanes 28-2970 66.8 C=C Alkenes -680 449. C-H Alkanes 3-470 02.2 C-O Alcohols, Ethers, 0-300 34. C-I Aliphatic iodo 0-0 472.39 S-S Polysulfides 470-0 Table 2: FT-IR Peak Values of Semecarpus anacardium Water Leaf Functional 3432.66 O-H Hydrogen bonded 30-30 90.2 Carbonyl Transition metal carbonyl 800-0 6.2 C=C Alkenes -680 Table 3: FT-IR Peak Values of Semecarpus anacardium Chloroform Leaf Functional Functional 387.83 - Unknown - 78.36 C=O Aldehydes, Ketones, 690-7 3733.70 - Unknown - 42.28 C-H Alkanes 3-470 84. O-H Hydrogen bonded 30-30 363.89 NO2 Nitro 300-370 24.3 O-H Monomeric Carboxylic 00-36 222.8 C-N Amines, Amides 80-3 acids 343.62 N-H Amines, Amides 3300-00 092.80 C-O Alcohols, Ethers, 0-300 3004.74 H-C Aromatic H-X group 3000-30 93.73 C-H Alkenes 67-99 2969.80 C-H Alkanes 28-2970 903.9 C-H Alkenes 67-99 29. C-H Alkanes 28-2970 773.00 C-Cl Aliphatic chloro 700-800 2. C C Alkynes 0-22 29.9 C-I Aliphatic iodo 0-0 cm - Table 4: FT-IR Peak Values of Semecarpus anacardium Toluene Leaf Functional 3839.6 - Unknown - 3649.93 O-H Monomeric Carboxylic acids 00-36 298.66 C-H Alkanes 28-2970 2364. - Unknown - 726.7 C=O Aldehydes, Ketones, 690-7.02 NO2 Nitro 300-370 88.34 C-N Secondary amine 30-90 979 C-H Alkenes 67-99 472.00 S-S Polysulfides 470-0 Table : FT-IR Peak Values of Semecarpus anacardium CCl4 Leaf Functional cm - Functional 3439.44 O-H Hydrogen bonded 30-30 970.8 C-H Alkenes 67-99 294.89 C-H Alkanes 28-2970 894.07 C-H Alkenes 67-99 297.37 C-H Alkanes 28-2970 829.73 C-H Alkenes 67-99 2849.26 C-H Alkanes 28-2970 729.82 C-H Aromatic rings 690-900 2329.08 - Unknown - 79. C-H Aromatic rings 690-900 736.08 C=O Aldehydes, Ketones, 690-7 646.63 C-Br Aliphatic bromo 0-700 4.9 NO2 Nitro 0-70.98 C-Br Aliphatic bromo 0-700 494.30 C=C-C Aromatic ring Aryl -4 78. C-I Aliphatic iodo 0-0 Stretch 462.86 C-H Alkanes 3-470.6 C-I Aliphatic iodo 0-0 377.66 C-H Alkanes 3-470 493.23 S-S Polysulfides 470-0 363.92 NO2 Nitro 300-370 483.02 S-S Polysulfides 470-0 27.76 C-O Alcohols, Ethers, 0-300 473.97 S-S Polysulfides 470-0

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 247.76 C-O Alcohols, Ethers, 0-300 464.0 S-S Aryl disulfides 430-0 87.96 C-N Amines, Amides 80-3 4.0 S-S Aryl disulfides 430-0 08. C-O Alcohols, Ethers, 0-300 - - - - Table 6: FT-IR Peak Values of Semecarpus anacardium Ethyl acetate Leaf Functional Functional 3630. O-H Monomeric Alcohols, 90-36.8 C=C Aromatic rings 0-0 Phenols 48.02 O-H Monomeric Carboxylic 00-36 46.86 C-H Alkanes 3-470 acids 3464.77 N-H Amines, Amides 3300-00 447.87 C-H Alkanes 3-470 298.3 H-O H-bonded H-X group 00-00 373.22 C-H Alkanes 3-470 2942.0 C-H Alkanes 28-2970 300.66 NO2 Nitro 300-370 2909.9 C-H Alkanes 28-2970 2 C-O Alcohols, Ethers, 0-300 2878.6 C-H Alkanes 28-2970.39 C-O Alcohols, Ethers, 0-300 2674.36 O-H Hydrogen bonded 00-2700 098. C-O Alcohols, Ethers, 0-300 Carboxylic acids 2473.39 - Unknown - 047.6 C-N Primary amines 0-090 2307.39 - Unknown - 004.6 C-F Aliphatic fluoro 000-2.0 C C Alkynes 0-22 938.3 C-H Alkenes 67-99 86. Carbonyl Transition metal 800-0 97.93 C-H Alkenes 67-99 carbonyl 09.89 Carbonyl Transition metal 800-0 847.4 C-H Aromatic rings 690-900 carbonyl 889.93 Carbonyl Transition metal 800-0 786.62 C-H Aromatic rings 690-900 carbonyl 743.93 C=O Aldehydes, Ketones, 690-7 634. C-Br Aliphatic bromo 0-700 6.7 C=C Alkenes -680 8. C-Br Aliphatic bromo 0-700 Table 7: FT-IR Peak Values of Semecarpus anacardium Hexane Leaf cm - Functional Frequency, cm - cm - Functional Frequency, cm - 34.66 N-H Amines, Amides 3300-00 87.3 C-N Amines, Amides 80-3 294.63 C-H Alkanes 28-2970 8.43 C-O 2924.63 C-H Alkanes 28-2970 080.94 C-O Alcohols, Ethers, Carboxylic acids, Alcohols, Ethers, Carboxylic acids, 0-300 0-300 2869.3 C-H Alkanes 28-2970 026.43 C-F Aliphatic fluoro 000-742. C=O Aldehydes, Ketones, 690-7 967.76 C-H Alkenes 67-99 626.28 C=C Alkenes -680 893.83 C-H Alkenes 67-99 492.33 NO2 Aromatic nitro in simple Hetero-oxy 48-88. C-H Alkenes 67-99 9.07 C-H Alkanes 3-470 700. C-H Aromatic rings 690-900 377.02 C-H Alkanes 3-470 646.7 C-Br Aliphatic bromo 0-700 363.7 C-H Alkanes 3-470 7. C-Br Aliphatic bromo 0-700 33.23 NO2 Nitro Compounds 300-370 497.39 S-S Polysulfides 470-0 248.70 C-O Alcohols, Ethers, Carboxylic acids, 0-300 - - - - 6

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 cm - Table 8: FT-IR Peak Values of Semecarpus anacardium Ethyl alcohol Leaf extract Functional Functional 3338.6 O-H Hydrogen bonded 30-30 380.7 C-H Alkanes 3-470 2974. H-O H- bonded H-X group 00-00 330.73 NO2 Nitro 300-370 2927.67 C-H Alkanes 28-2970 274.67 C-O Alcohols, Ethers, 0-300 288.4 C-H Alkanes 28-2970 090.48 C-O Alcohols, Ethers, 0-300 4.90 O-H Hydrogen bonded 00-2700 0.43 C-O Alcohols, Ethers, 0-300 Carboxylic acids 26.37 C C Alkynes 0-22 88.27 C-H Aromatic rings 690-900 9.32 Carbonyl Transition metal 800-0 803.7 C-H Alkenes 67-99 carbonyl 64.8 C=C Alkenes -680 77.9 C-H Alkenes 67-99 4.62 C-H Alkanes 3-470 - - - - 422.86 C-H Alkanes 3-470 - - - - Table 9: FT-IR Peak Values of Semecarpus anacardium Methanol Leaf Functional 38.39 - Unknown - 3366.30 N-H Amines, Amides 3300-00 2946.08 C-H Alkanes 28-2970 2833.7 C-H Methoxy, Methyl ether O-CH3 28-28 22.92 O-H Hydrogen bonded Carboxylic acids 00-2700 2223.33 C C Alkynes 0-22 44. Carbonyl Transition metal carbonyl 800-0 6.62 C=C Alkenes -680 449.68 C-H Alkanes 3-470 29.47 C-O Alcohols, Ethers, 0-300 4.64 C-O Alcohols, Ethers, 0-300 03.99 Methylene >CH2 Cyclohexane ring vibrations 000-0 Table 0: FT-IR Peak Values of Semecarpus anacardium Acetone Leaf Functional 342.46 O-H Hydrogen bonded 30-30 2978.67 H-O H- bonded H-X group 00-00 3.29 C C Alkynes 0-22 70.7 C=O Aldehydes, Ketones, 690-7 6.7 C=C Alkenes -680 42. C-H Alkanes 3-470 368.6 NO2 Nitro 300-370 232. C-N Amines, Amides 80-3 63.09 C-O Alcohols, Ethers, 0-300 093.66 C-O Alcohols, Ethers, 0-300 022.6 Methylene >CH2 Cyclohexane ring vibrations 000-0 9.4 C-H Alkenes 67-99 Table : FT-IR Peak Values of Semecarpus anacardium 2-propanol Leaf Functional Functional 388.0 - Unknown - 768.93 C=O Open chain Acid 7-790 anhydrides 3798. - Unknown - 646.3 C=C Alkenes -680 3734.9 - Unknown - 467.73 C-H Alkanes 3-470 336.96 N-H Amines, Amides 3300-00 9.7 C-H Alkanes 3-470 297.3 H-O H- bonded H-X group 00-00 379.8 C-H Alkanes 3-470 2932.69 C-H Alkanes 28-2970 34.03 C-H Alkanes 3-470 2884.32 C-H Alkanes 28-2970 309. NO2 Nitro 300-370 272.64 C-H Aldehyde 2700-2800 6.90 C-O Alcohols, Ethers, 0-300 269.32 O-H Hydrogen bonded 00-2700 29.90 C-O Alcohols, Ethers, 0-300 Carboxylic acids 29.24 - Unknown - 0.44 C-O Alcohols, Ethers, 0-300 29.69 C C Alkynes 0-22 93.02 C-H Alkenes 67-99 903.2 Carbonyl Transition metal carbonyls 800-0 87.3 C-H Alkenes 67-99 62

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 Table 2: FT-IR Peak Values of Semecarpus anacardium Petroleum ether Leaf Functional 3438.97 O-H Hydrogen bonded 30-30 297.9 C-H Alkanes 28-2970 2849.3 C-H Alkanes 28-2970 737.22 C=O Aldehydes, Ketones, 690-7 463.08 C-H Alkanes 3-470 384.3 NO2 Nitro 300-370 27.7 C-N Amines, Amides 80-3 87.83 C-N Amines, Amides 80-3 08.9 C-O Alcohols, Ethers, 0-300 97.02 C-H Alkenes 67-99 894.90 C-H Alkenes 67-99 473.4 S-S Polysulfides 0-470 Table 3: FT-IR Peak Values of Semecarpus anacardium 2- butanone Leaf Functional Functional 84.29 O-H Monomeric Carboxylic 00-36 47.2 C-H Alkanes 3-470 acids 24.3 O-H Hydrogen bonded 30-30 366. C-H Alkanes 3-470 346.2 N-H Amines, Amides 3300-00 7.24 C-O Alcohols, Ethers, 0-300 2980.9 H-O H-bonded H-X group 00-00 6.4 C-O Alcohols, Ethers, 0-300 29.34 C-H Alkanes 28-2970 72.93 C-O Alcohols, Ethers, 0-300 296 C-H Alkanes 28-2970 086.89 C-O Alcohols, Ethers, 0-300 2883.0 C-H Alkanes 28-2970 996.2 P-O-C Aliphatic Phosphates 990-0 222.9 C C Alkynes 0-22 9.67 C-H Alkenes 67-99 78.83 C=O Aldehydes, Ketones, 690-7 7.62 C-H Aromatic rings 690-900 9.4 C-H Alkanes 3-470 88.7 C-I Aliphatic iodo 0-0 Table 4: FT-IR Peak Values of Semecarpus anacardium Dichloromethane Leaf Functional 3438.08 O-H Hydrogen bonded 30-30 2923.26 C-H Alkanes 28-2970 282.3 C-H Alkanes 28-2970 736.00 C=O Aldehydes, Ketones, 690-7 4.92 C-H Alkanes 3-470 379.03 C-H Alkanes 3-470 62.94 C-O Alcohols, Ethers, 0-300 97.96 C-H Alkenes 67-99 73. C-H Alkenes 67-99 Table : FT-IR Peak Values of Semecarpus anacardium Ethyl ether Leaf extract Functional 3332.3 O-H Hydrogen bonded 30-30 2924.66 C-H Alkanes 28-2970 283.66 C-H Alkanes 28-2970 706.2 C=O Aldehydes, Ketones, 690-7 66.46 C=C Alkenes -680.38 NO2 Nitro 0-70 8.8 C-H Alkanes 3-470 377. C-H Alkanes 3-470 26.68 C-N Amines, Amides 80-3 97.69 C-H Alkenes 67-99 829.9 C-H Alkenes 67-99 78.78 C-H Alkenes 67-99 Table 6: FT-IR Peak Values of Semecarpus anacardium Stem Powder Functional 3330. O-H Hydrogen bonded 30-30 298.66 C-H Alkanes 28-2970 734.07 C=O Aldehydes, Ketones, 690-7 63

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 cm - 67.7 C=C Alkenes -680 9.06 C-H Alkanes 3-470 39.7 NO2 Nitro Compounds 300-370 2.9 C-N Amines, Amides 80-3 063.94 C-O Alcohols, Ethers, 0-300 33.90 C-I Aliphatic iodo 0-0 Table 7: FT-IR Peak Values of Semecarpus anacardium Methanol Stem Bark Functional 387.06 - Unknown - 3366. O-H Hydrogen bonded 30-30 29.34 C-H Alkanes 28-2970 2833.7 C-H Methoxy, Methyl ether O-CH3 28-28 22.84 O-H Hydrogen bonded Carboxylic acids 00-2700 2226.6 C C Alkynes 0-22 44. Carbonyl Transition metal carbonyl 800-0 64.7 C=C Alkenes -680 6 C-H Alkanes 3-470 4.90 C-O Alcohols, Ethers, 0-300 032.44 Methylene >CH2 Cyclohexane ring vibrations 000-0 Table 8: FT-IR Peak Values of Semecarpus anacardium 2- butanone Stem Bark Functional Frequency, cm - cm - Functional Frequency, 29.9 O-H Hydrogen bonded 30-30 9.3 C-H Alkanes 3-470 346.79 O-H Hydrogen bonded 30-30 46.78 C-H Alkanes 3-470 2980.23 H-O H-bonded H-X group 00-00 36.37 NO2 Nitro 300-370 29. C-H Alkanes 28-2970 7.3 C-N Amines, Amides 80-3 297 C-H Alkanes 28-2970 6.43 C-O Alcohols, Ethers, 0-300 Carboxlic acids, 2883.27 C-H Alkanes 28-2970 72.63 C-O Alcohols, Ethers, 0-300 Carboxlic acids, 2830.86 C-H Methoxy, Methyl 28-28 086.89 C-O Alcohols, Ethers, 0-300 ether O-CH3 Carboxlic acids, 6.77 O-H Hydrogen bonded 00-2700 033.84 C-N Primary amine 0-090 Carboxylic acids 2302.49 - Unknown - 996.44 P-O-C Aliphatic 990-0 Phosphates 222.90 C C Alkynes 0-22 9.37 C-H Alkenes 67-99 887.2 Carbonyl Transition metal 800-0 7.89 C-H Aromatic rings 690-900 carbonyl 78.77 C=O Aldehydes, Ketones, Carboxylic acids, 690-7 89.32 C-I Aliphatic iodo cm - 0-0 84.0 82 80 62.3 78 76 74 72 70 68 30 90.2,34. 66 64 62 299.69,.8 8 34.,6.48 449.,8.30 472.39,.38 33,8.27 6 66.8,6.6 4.3 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig :FT-IR Spectrum of Semecarpus anacardium Leaf Powder 0 3432.66,.9 6.2,.9 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 2:FT-IR Spectrum of Water Semecarpus anacardium Leaf 64

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 63.2 9.9 387.83,6.03 3733.70,.34 2.,6.9 93.73,9.23 903.9,4.7 773.00,8. 90 29.,46.92 8 2969.80,43.62 092.80,43.9 30 343.62,39.04 84.,36.98 3004.74,37.8 24.3,37.02 42.28,26. 80 7 70 646.7,83 742.,78.3 700.,79.04 497.39,78.47 626.28,78.43 026.43,76.88 88.,77.0 7.,79.69 33.23,76.00 893.83,77. 248.70,7.70 8.43,74. 492.33,72.76 377.02,7.06 34.66,70. 363.7,72.93 87.3,70.48 080.94,70.38 0 6 9.07,67.44 967.76,67.27 363.89,9. 78.36,7.8 222.8,9.76 29.9,4.89 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 3:FT-IR Spectrum of Chloroform Semecarpus anacardium Leaf 00.46 0 99. 99.0 98. 98.0 97. 97.0 96. 3839.6,96.82 96.0 2364.,96.0 9. %R 9.0 3649.93,94.06 298.66,93.92 94. 726.7,94.8.02,94.76 472.00,94.73 94.0 88.34,94.70 93. 979,93.6 93.0 92. 92.0 2869.3,63.44 294.63,6.88 2924.63,4.06 3.8 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 7:FT-IR Spectrum of Hexane Semecarpus anacardium Leaf 62.9 26.37,.34 9.32,8.4 64.8,.76 4.90,.9 803.7,.83 77.9,42.6 274.67,.27 330.73,34.39 30 422.86,27.62 4.62,26.8 380.7,22.29 9. 9.0 90. 0 2927.67,.66 3338.6,2.02 2974.,.98 288.4,9.76 090.48,4.74 0.43,0.89 88.27,3.6 90 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 4:FT-IR Spectrum of Toluene Semecarpus anacardium Leaf 3. 30 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 8:FT-IR Spectrum of Ethyl alcohol Semecarpus anacardium Leaf 6.0 2329.08,29.3 38.39,8. 4.9,22.44 78.,22.3 646.63,.69 2223.33,.97 0 0 %R 3439.44,8.9 736.08,04.8 829.73,9.02.98,2.6.6,2.23 894.07,6.2 493.23,9.86 483.02,.78 473.97,9.23 494.30,.6 729.82,2.88 464.0,.2 27.76,0.30 79.,2.06 4.0,2. 363.92,09. 08.,08.49 377.66,07.86 247.76,0.30 970.8,06.70 87.96,06.06 30 22.92,.67 44.,.02 00 462.86,00.86 29.47,29.0 9 90 294.89,90.8 6.62,8.72 8 0 80 2849.26,83.6 2833.7,0.9 3366.30,0.27 2946.08,0.7 449.68,3.87 4.64,9.26 03.99,0.47.6 77.4 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig :FT-IR Spectrum of CCl4 Semecarpus anacardium Leaf 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 9:FT-IR Spectrum of Methanol Semecarpus anacardium Leaf 62.8 2674.36,.24 2307.39,6.36 2473.39,4.92.0,.7 09.89,4.09 86.,48.96 3.29,.88 889.93,.70.8,43.93 63.09,4.7 30 3630.,34.63 48.02,34.7 3464.77,32.72 2878.6,29.79 6.7,3.93 97.93,33.33.39,3.98 786.62,3.86 004.6,30.73 30 2978.67,36. 42.,30. 9.4,39.0 093.66,36.72 022.6,.38 2909.9,6.9 938.3,9.64 847.4,8.27 232.,23.48 368.6,.3 0 2942.0,.29 298.3,2.64 743.93,.32 46.86,0.2 2,.23 373.22,.2 098.,0. 447.87,7.9300.66,7.70 047.6,.3 634.,4.62 8.,.6 0 342.46,3.33 6.7,3.84 70.7,.96 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 6:FT-IR Spectrum of Ethyl acetate Semecarpus anacardium Leaf 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 0: FT-IR Spectrum of Acetone Semecarpus anacardium Leaf 6

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 67.0 6 388.0,62.77 3798.,62.64 3734.9,6.8 903.2,6.07 29.24,64. 29.69,64.29 768.93,6.43 272.64,.22 269.32,.90 646.3,46.64 07.9 0 00 9 90 30 34.03,3.48 9.7,32 %R 8 80 0 336.96,.92 2884.32,.39 2932.69,8.84 297.3,0.8 467.73,.08309.,. 379.8,2.4 0.44,7.2 6.90, 29.90,8.2 93.02,2.9 87.3,9.4 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig :FT-IR Spectrum of 2-propanol Semecarpus anacardium Leaf 24.3 23 22 2 9 8 7 6 %R 4 3 2 3438.97,4.76 737.22,4. 463.08,3.32 27.7,.78 87.83,3.69 08.9,4.68 894.90,7.8 97.02,3.33 473.4,3.43 7 70 6 63.2 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig :FT-IR Spectrum of Ethyl ether Semecarpus anacardium Leaf 93.9 92 90 88 86 84 82 80 78 76 74 72 70 2924.66,64.83 706.2,7.6 26.68,77. 0 09 08 2849.3,09.8 384.3,.69 68 66 64 62 298.66,64.6 734.07,67.3 9.06,63.79 07 06 04.6. 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 2:FT-IR Spectrum of Petroleum ether Semecarpus anacardium leaf 2.9,6.90 39.7,.99 8 3330.,.22 67.7,9.92 33.90,.7 6.4 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 6:FT-IR Spectrum of Semecarpus anacardium Stem Powder 6.9 387.06,9. 2226.6,3. 222.9,4.62 44.,47.2 996.2,49.72 7.24,47.3 086.89,42.7 7.62,.02 30 22.84,.4 64.7,33.38 30 84.29,33.3 24.3,32.4 346.2,32.9 6.4,34.79 9.67,34.0 2883.0,27.6 296,24.96 0 29.34,6.79 2980.9,9.97 78.83,3.9 9.4,9.9 47.2,.77 366.,6.7 72.93,7.2 88.7,2.04 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 3:FT-IR Spectrum of 2- butanone Semecarpus anacardium leaf 0.6 0 00 0 2833.7,2. 3366.,.78 29.34,2.07 6,.3 4.90,. 032.44,.84 0 30 30 2800 20 00 800 0 0 0 000 800 0 69.2 Fig 7:FT-IR Spectrum of Methanol Semecarpus anacardium Stem Bark 6 6.77,6. 2302.49,66.3 222.90,63.0 887.2,6.49 99 98 97 96 9 94 %R 93 92 9 3438.08,96.34 4.92,93.09 379.03,92.93 62.94,9.98 97.96,93.6 73.,9. 30 0 346.79,32.37 29.9,26.87 2830.86,44.67 2883.27,7.07 297,4.42 29.,8.2 2980.23,.6 78.77,3.84 7.3,44. 996.44,.28 7.89,39.89 086.89,34.07 033.84,3.4 6.43,. 9.37,23.73 89.32,.87 36.37,4.80 72.63,. 9.3,0.29 46.78,8.26 90 89 88 86.7 282.3,90.27 736.00,94 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 4:FT-IR Spectrum of Dichloromethane Semecarpus anacardium Leaf 0 30 30 2800 20 00 800 0 0 0 000 800 0 Fig 8:FT-IR Spectrum of 2- butanone Semecarpus anacardium Stem Bark The functional groups which are present in the samples are further described in details. C-H bond of Alkane (28-2970 cm - ) group frequency are only present only in crude leaf powder of Semecarpus anacardium and in chloroform, toluene, carbon tetrachloride and 66

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 hexane leaf extracts. C-H bond of Alkane (3-470 cm - ) group frequency were found to be present only in Semecarpus anacardium acetone leaf extract. Both the group frequencies of the C-H bond of Alkane (28-2970 cm - ) and (3-470 cm - ) were present in ethyl acetate, ethyl alcohol, methanol, 2-propanol, petroleum ether, 2-butanone, dichloromethane, ethyl ether leaf extracts, stem powder and methanol, 2- butanone stem bark extracts. C=C bond of Alkene (-680 cm - ) group frequency were present in the crude Semecarpus anacardium leaf powder and water, ethyl acetate, ethyl alcohol, methanol, acetone, 2-propanol, ethyl ether leaf extracts, stem powder and methanol extract of stem bark. C-H bond of Alkene (67-99 cm - ) group frequency occured in chloroform, toluene, carbon tetrachloride, ethyl acetate, hexane, ethyl alcohol, acetone, 2-propanol, petroleum ether, 2-butanone, dichloromethane, ethyl ether leaf extracts and 2- butanone stem bark extract. C C bond of Alkyne (0-22 cm - ) group frequency was present in chloroform, ethyl acetate, ethyl alcohol, methanol, acetone, 2- propanol, 2-butanone leaf extracts and methanol and 2- butanone stem bark extracts. C-H bond of Aromatic ring (690-900 cm - ) group frequency were present in carbon tetrachloride, ethyl acetate, hexane, ethyl alcohol, 2- butanone leaf extracts and 2- butanone stem bark extract. C=C bond of Aromatic ring (0-0 cm - ) group frequency were present only in ethyl acetate leaf extract. O-H bond of Monomeric (90-36 cm - ) group frequency were present only in ethyl acetate leaf extract. O-H bond of Hydrogen bonded (30-30 cm - ) group frequency were present in leaf powder and water, chloroform, carbon tetrachloride, ethyl alcohol, acetone, petroleum ether, 2- butanone, dichloromethane, ethyl ether leaf extracts, stem powder and methanol, 2- butanone stem bark extracts. O-H bond of Monomeric Carboxylic acid (00-36 cm - ) group frequency was present in chloroform, toluene and 2- butanone leaf extracts. O-H bond of Hydrogen bonded Carboxylic acid (00-2700 cm - ) group frequency were present in ethyl acetate, ethyl alcohol, methanol, 2- propanol leaf extracts and methanol, 2- butanone stem bark extracts. N-H bond of Amines, Amides (3300-00 cm - ) group frequency were present in chloroform, ethyl acetate, hexane, methanol, 2- propanol and 2- butanone leaf extracts. C-N bond of Amines, Amides (80-3 cm - ) group frequency were present in chloroform, carbon tetrachloride, acetone, petroleum ether and ethyl ether leaf extracts, stem powder and 2- butanone stem bark extract. C-O bond of Alcohols, Ethers, (0-300 cm - ) group frequency were present in leaf powder and chloroform, carbon tetrachloride, ethyl acetate, hexane, ethyl alcohol, methanol, acetone, 2-propanol, petroleum ether, 2- butanone and dichloromethane leaf extracts, stem powder and methanol, 2- butanone stem bark extracts. C=O bond of Aldehydes, Ketones, (690-7 cm - ) group frequency were present in chloroform, toluene, carbon tetrachloride, ethyl acetate, hexane, acetone, petroleum ether, 2-butanone, dichloromethane and ethyl ether leaf extracts, stem powder and 2- butanone stem bark extract. NO2 bond of Nitro (0-70 cm - ) group frequency were present only in carbon tetrachloride and ethyl ether leaf extracts. NO2 bond of Nitro (300-370 cm - ) group frequency were present in chloroform, toluene, carbon tetrachloride, ethyl acetate, hexane, ethyl alcohol, acetone, 2-propanol and petroleum ether leaf extracts, stem powder and 2- butanone stem bark extract. The C-H vibrations for methyl and methylene are the most characteristic in terms of recognizing the compound as an organic compound containing at least one aliphatic fragment. In this functional groups from Saturated Aliphatic and Alicyclic like Cyclohexane ring vibrations (000-0 cm - ) were present in methanol and acetone leaf extracts and methanol stem bark extract. Special methyl ( CH3) frequencies like Methoxy, methyl ether O-CH3, C-H (28-28 cm - ) were present only in 2- butanone stem bark extract.the existence of one or more aromatic rings in a structure is normally readily determined from the C-H and C=C-C ring-related vibrations. The other most important set of bands are the aromatic ring vibrations centered around 0 and 0 cm -, which usually appear as a pair of band structures, often with some splitting. The appearance and ratio of these band structures is strongly dependent on the position and nature of substituents on the ring. Aromatic ring (aryl) group C=C-C (4- cm - ) was present only in carbon tetrachloride leaf extract. In Halogenated aliphatic, the C-X bond typically possesses a unique group frequency, which may be assigned to the halogen carbon ing. Aliphatic fluoro (000- cm - ) C-F were present in ethyl acetate and hexane leaf extracts. Aliphatic chloro (700-800 cm - ) C-Cl were present only in chloroform leaf extract. Aliphatic bromo (0-700 cm - ) C-Br were present in carbon tetrachloride, ethyl acetate and hexane leaf extracts. Aliphatic iodo (0-0 cm - ) C-I were present in leaf powder and chloroform, carbon tetrachloride, 2-butanone leaf extracts, stem powder and 2- butanone stem bark extract. In Amino Compounds with some respects, the infrared spectra and the characteristic group frequencies of amines tend to parallel those of alcohols and ethers. The terms primary, secondary, and tertiary are used to describe amines, but the substitution relates to the nitrogen, not the adjoining carbon (as with alcohols). Only the primary and secondary amines exhibit the most characteristic group frequencies, which are associated with the N-H bond. Primary amine (0-090 cm - ) CN were present in ethyl acetate leaf extract and 2- butanone stem bark extract. Secondary amine (30-90 cm - ) CN were present only in toluene leaf extract. The Carbonyl group are not only chemically important, but are also important in the interpretation of infrared spectra. The C=O absorption is almost always one of the most characteristic in the entire spectrum, and it is also most likely to be the most intense spectral feature. Aldehyde (2700-2800 cm - ) group frequencies were present only in 2-propanol leaf extract. Open-chain acid anhydride (7-790 cm - ) group frequencies were present in only in 2- propanol leaf extract. Transition metal carbonyls (800-0 cm - ) group frequencies were present water, ethyl acetate, ethyl alcohol, 2-propanol leaf extracts and methanol, 2-butanone stem bark extracts. The group of Hetero-oxy features X-O (where X= nitrogen, sulfur, phosphorus, and silicon) and X=O vibrations. In general, the ing frequencies is observed within the main fingerprint spectral region (0 0 cm - ). Many of the can be considered to be analogs of ethers, especially when an alkoxy group is present, featuring the X-O-C linkage. Aromatic nitro (48- cm - ) group frequencies were present only in hexane leaf extract. Aliphatic phosphates (990-0 cm - ) P-O-C were present in 2- butanone leaf extract and 2- butanone stem bark extract. Thiols and Thio-substituted by definition, can be considered to be the direct analogs of the equivalent oxygenated such as alcohols and ethers. Catenation is a unique chemical characteristic of sulfur, where the formation of S-S bonds in extended chains is common, hence the inclusion here of the S-S ing frequencies. Aryl disulfides (430-0 cm - ) S-S was present only in CCl4 leaf extract. Polysulfides (470-0 cm - ) S-S was present in leaf powder and toluene, carbon tetrachloride, hexane, petroleum ether leaf extracts. In H-X group, the ing of bonds between heavier atom and hydrogen (H-X-) occurs in the region between 00 and 00cm -. The X-atoms and the structure to which they are attached determines the exact frequency. H-C (aromatic) (3000-30 cm - ) group frequency was present only in chloroform leaf extract. H- bonded (H-O): (00-00 cm - ) group frequencies were present in ethyl acetate, ethyl alcohol, acetone, 2-propanol, 2-butanone soxhlet leaf extracts and 2- butanone stem bark extract. The above listed infrared functional group absorptions characteristic were cited from the literature 4-6. The FT-IR analysis 67

Asian J Pharm Clin Res, Vol 6, Suppl, 3, 9-98 revealed the similarity and variation between the leaf and stem parts of Semecarpus anacardium. From the spectra we can clearly see that although they show substantial overlap of each absorption spectrum of various components, each band represents an overall overlap of some characteristic absorption peaks of functional groups in the samples. By using the macroscope fingerprint characters of FT-IR spectrum, we can judge the origin of different extracts accurately and effectively, trace the constituents in the extracts, identify the medicinal materials and even evaluate the qualities of medicinal materials 7.. Skoog DA, Holler JF, Nieman TA. Principles of Instrumental Analysis. In: Applications of Infrared spectrometry. th ed. Bangalore: Thomson Brooks; 0. p. 4-428. 6. Kealey D, Haines PJ. Instant Notes: Analytical Chemistry. In: Infrared and Raman Spectrometry: Principles and Instrumentation. Oxford: Bios Scientific Publishers Limited, 02. p. 233-24. CONCLUSION The FT-IR analysis of Semecarpus anacardium clearly reveals the differences of categories of chemical constituents in crude powder and 4 different polar and nonpolar organic solvent extracts of leaf and stem bark. The crude plant material when extracted by organic solvents, shows more characteristic features in the FT-IR spectrum as the chemical compositions become more concentrated. Due to the similarity of the extraction method i.e. soxhlet extraction, the chemical constituents in the concentrated extracts are relatively consistent and therefore they show higher repeatability and comparability in FT-IR spectra. So, FT-IR spectrum reflecting objectively the panorama of chemical composition in complex plant material is a most appropriate method to validate and identify the mixed substance products such as traditional herbal medicine as well as their corresponding extracts. The results of the present study confirms that leaf and stem bark of Semecarpus anacardium is rich in the resources of phytoconstituents. REFERENCES. Liu H, Sun S, Lv G, Chan KKC. Study on Angelica and its different extracts by Fourier transform infrared spectroscopy and twodimensional correlation IR spectroscopy. Spectrochimica Acta Part A 06; 64: 32 326. 2. Griffiths PR, De Haseth JA. Fourier transform infrared spectroscopy. New York: Wiley, 986. 3. Bobby MDN, Wesely EG, Johnson M. FT-IR Studies on the leaves of Albizia Lebbeck Benth. Int J Pharm Pharm Sci 2; 4(3): 293-296. 4. Cheng CG, Liu J, Wang H, Xiong W. Infrared spectroscopic studies of Chinese medicines. Appl. Spectros. Rev 0; : 6 78.. Bunaciu AA, Aboul-Enein HY, Fleschin S. Recent applications of Fourier transform infrared spectrophotometry in herbal medicine analysis. Appl. Spectros. Rev ; 46(3): 2. 6. Bunaciu AA, Aboul-Enein HY, Fleschin S. Application of Fourier transform infrared spectrophotometry in pharmaceutical drugs analysis. Appl. Spectros. Rev 0; (3): 6 29. 7. Mariswamy Y, Gnanaraj WE, Antonisamy JM. FT-IR Spectroscopic Studies on Aerva Lanata (L.) Juss. Ex Schult. Asian J Pharm Clin Res 2; (2): 82-86. 8. Khare C.P. Indian medicinal plants: An Illustrated Dictionary. C.P. Khare ed. Verlag Berlin / Heidelberg: Springer; 07.p.97. 9. Kirthikar KR, Basu BD. Indian medicinal plants. Dehra Dun, India: Publisher Bishen Singh Mahendra Pal Singh; 933.p.667 669. 0. Murthy SSN. New bioflavonoid from Semecarpus anacardium Linn. Clin Acta Turcica 992; : 33 37.. Premalatha B. Semecarpus anacardium Linn. nuts - a boon in alternative medicine. Indian J Exp Biol 00; 38(2): 77 82. 2. Ramprasath VR, Shanthi P, Sachdanandam P. Immunomodulatory and Anti-inflammatory effects of Semecarpus anacardium Linn. nut milk extract in experimental inflammatory conditions. Biol. Pharm. Bull 06; 29(4): 693-700. 3. re VN, Nathar VN. Phytoconstituents, Proximate and Mineral Composition of Semecarpus anacardium L. An Ethnomedicinal Plant. International Journal of Current Research ; 3(6): 423-428. 4. Coates J. Encyclopedia of Analytical Chemistry. In: Interpretation of Infrared Spectra, A Practical Approach. R.A. Meyers ed. Chichester: John Wiley & Sons Ltd; 00. p. 08-0837. 68