International Journal of Innovative Pharmaceutical Sciences and Research www.ijipsr.com MOLECULAR STRETCHES OF ACTIVE PRINCIPLES ISOLATED FROM TUBERS AND SEEDS OF Gloriosa superba THROUGH FTIR- SPECTROSCOPIC ANALYSIS 1 S.Megala*, 2 R.Elango 1 Project Fellow, Faculty of Science, Department of Microbiology, Annamalai University, INDIA 2 Assistant professor, Department of Microbiology, Faculty of Agriculture, Annamalai Nagar, Tamil Nadu, INDIA Abstract In the present study FTIR spectroscopic analysis tubers and seeds extracts of Gloriosa superba, the different peaks obtained in the functional group region was in between 4000-1300 cm -1. It is inferred that the tubers and seeds extract revealed that the peaks are involved in inter molecular hydrogen bonding, N-H bond either primary, secondary or tertiory amines, carbonyl group (> C=O), which may be either with the form of carbonyl acid or carbonyl group coupled with C = C group. Certain peaks represent alcoholic (O-H) group also. Keywords: Medicinal plant, Gloriosa superba, FTIR and Plant extract. Corresponding Author: S. Megala Department of Microbiology Annamalai University, INDIA Email: megala@scientist.com Phone: +91 9500259389 Available online: www.ijipsr.com July Issue 1289
INTRODUCTION Gloriosa superba L. is a valuable tropical medicinal plant. All its parts find diverse use in indigenous system of medicine. Gloriosa is highly valued in both traditional and modern therapies. Its tubers and seeds (Active content of colchicine) are use mainly for treating gout and rheumatisms. Gloriosa is used for treating a wide range of human ailments throughout the tropics. In, India, the auyrvedic pharmacopoeia recommends G. superba as an ecbolic inlabour, purgative, an anthelmintic and cure against leprosy, colic, chronic ulcers, haemorrhoids, skin parasites, head lice and tumors [1-6]. FTIR has been proven to be useful in studying compositional changes in plant cell walls during development. Therefore, it can possibly be used to determine changes in cell wall architecture upon exposure to organic contaminants [7]. Initially, the use of infrared spectroscopical method is restricted only for structural elucidation of isolated compounds from the herbal matrices. It was also found to be useful in phytochemical studies as a fingerprinting device, for comparing natural with synthetic sample [8]. Drug discovery from the medicinal plants continues to provide new and important leads against various pharmacological targets including cancer, HIV/AIDS, Alzheimer s, malaria, infections and pain [9]. Adopt the approach of metabolic fingerprinting through the use of Fourier Transform Infrared technique (FTIR) to understand the composition, chemical structure and discrimination of biomolecules in medicinal plants of Tephrosia tinctoria and Atylosia albicans. IR spectrum in mid infrared region (4000 400cm -1 ) was used for discriminating and indentifying various functional groups present in two different species of medicinal plants [10]. The presence of the phytocompound (i.e.) Colchicine and other chemical constituents present in three different treated tuber and seed samples. The three different treated samples are Control (T 1 ), Chemical Fertilizer (T 2 ), and Organic Manure (T 3 ) of Glory Lily (Gloriosa superba) was confirmed using FTIR [11]. Available online: www.ijipsr.com July Issue 1290
MATERIALS AND METHODS (i) Sample preparation Tubers and seeds of Gloriosa superba plant materials taken for this study was shade dried individually at room temperature for two weeks and then powdered by using electric blender then, sieved the powder individually by using a nylon sieve in order to remove the large particles of samples. Collect a fine powder of sample in screw- cap bottle and stored for further use. (ii) Analysis of active principle compounds in the Gloriosa superba L. The qualitative analysis of the active principle of the G. superba performed by Fourier Transmission Infra-Red (FTIR) method as described by 12 (iii) Preparation of KBr discs The FT-IR spectra were recorded in a Thermo Nicolet, AVATAR 330 FT-IR system, Madison WI 53711-4495. The air dried tubers and seeds sample was grounded with a purified potassium bromide salt to remove scattering effects from large crystals. This powdered mixture is then pressed in a mechanical press to form a translucent pellet through which the beam of the spectrometer can be passed. (iv) Infra-Red analysis The frequency of the spectra set to analysis was between 4000 500 / cm wave number and the vibration spectrum were recorded as graphical chart. The instrument used to FTIR analysis was instrumentation lab of CAS in Marine Biology, Annamalai University, Parangipaettai. Available online: www.ijipsr.com July Issue 1291
TABLE 1 Molecular stretches of active principles isolated from Gloriosa superba tubers through FTIR - Spectroscopic analysis SI. NO Wave NO.(cm -1 ) Bond type (Stretching frequencies) 1. 526.57 C X 2. 576.72 C X 3. 665.44 C X (Alkynes) 4. 759.95 N O (Aromatic) 5. 854.47 C X (Aromatic) 6. 1018.41 C X, C O 7. 1157.29 C X, C O 8. 1244.09 C X, C O 9. 1419.61 N O 10. 1645.28 C C, N O 11. 2353.16 N H 12. 2926.01 N H 13. 3427.51 O H TABLE 2 Molecular stretches of active principles isolated from Gloriosa superba seeds through FTIR Spectroscopic analysis SI. NO Wave NO.(cm -1 Bond type ) (Stretching frequencies) 1. 464.84 C X 2. 601.79 C X 3. 667.37 C X (Alkenes) 4. 881.47 C O O C 5. 1033.85 C O ; C X 6. 1253.73 C O ; C X 7. 1386.82 C X 8. 1440.83 C C, N O 9. 1633.71 C = C 10. 1730.15 C = O 11. 2353.16 N- H 12. 2862.36 N H 13. 2924.09 N H 14. 3873.06 O H Available online: www.ijipsr.com July Issue 1292
RESULT AND DISCUSSION Fig. 1: Molecular stretches of active principles isolated from tuber of Gloriosa superba through FTIR - Spectroscopic analysis FTIR spectroscopic analysis of Gloriosa superba tubers An experiment was contacted to study the FTIR spectroscopic analysis of Gloriosa superba tubers. The results of the experiment a presented in (Table-1) (Fig-1). Infrared spectroscopic analysis given in the information about the possible functional groups of active principles between the wave number 4500 500 cm -1. Solid FTIR spectroscopic analysis using kbr disc method was employed to analyse the functional groups of the tuber in G. superba. The IR spectral analysis of tuber in G. superba revealed the following. The results of different peaks obtained in the functional group region (4000 to 1300 cm -1 ), it is inferred that, the broad band at 3000 3500 cm -1 may be due to OH group (involved in inter molecular hydrogen bonding). The peak at 2926 cm -1 may be due to N= N bond. The peak at 1645 cm -1 represents a carboxyl group (> C = O) which may be in the form of carboxyl acid (- C - OH) or a carboxyl group in conjugation with C = C-. The peak at 1419 cm -1 may be due to N O group. The peak at 1244 cm -1 C O group. The peak at 1157 cm -1 C O group. Available online: www.ijipsr.com July Issue 1293
Fig. 2: Molecular stretches of active principles isolated from seed of Gloriosa superba through FTIR Spectroscopic analysis FTIR spectroscopic analysis of Gloriosa superba seeds An experiment was contacted to study the FTIR spectroscopic analysis of Gloriosa superba seeds. The results of the experiment a presented in (Table-2) (Fig-2). Infrared spectroscopic analysis given in the information about the possible functional groups of active principles between the wave number 4500-500 cm -1. Solid FTIR spectroscopic analysis using kbr disc method was employed to analyse the functional groups of the seed in G. superba. The IR spectral analysis of tuber in G. superba revealed the following. The result of different peaks obtained in the functional group region (4000 to 1300 cm -1 ), it is inferred that, The broad band at 3000 3500 cm -1 may be due to OH group (involved in inter molecular hydrogen bonding). The peak at 2924 cm -1, 2862 cm -1, 2353 cm -1 may be due to N H bond. The peak at 1730 cm -1, 1633 cm -1, 1440 cm -1 may be due to C = O, C =C, C-C bond respectively. CONCLUSION From the study it can be concluded that the FTIR analysis revealed that based on the skeletal form of molecular stretches of active principles in amines or carbonyl or ketone or alcoholic groups of compounds were they mainly present in Gloriosa superba. The active principle derivatives are varied in tubers and seeds of Gloriosa superba. Available online: www.ijipsr.com July Issue 1294
ACKNOWLEDGEMENT The authors are thanks to University Grants Commission (UGC), New Delhi, for providing financial assistance under Major Research Project for this medicinal plant (F.NO.41-1182/2012 (SR). REFERENCE 1. Bunyapraphatsara N, Van Valken burg. Gloriosa superba L. In: de paduda LS, Bunyapraphatsara N, Lemmens RHMJ (eds). Plant Resources of South-East Asia No. 12(1) Medicinal and poisonous plants 1. Backhuys Publisherrs, Leiden, Netherlands. pp. 1999; 289 292. 2. Geetha S, Poornima S, Vaseegari J. Studies on the ethnobotany of rulars of Anaikatty hills, Coimbatore District. Coll Sci India, 2007; 1: 1-20. 3. Jagtap SD, Deokule SS, Bhosle SV. Some unique ethno medicinal uses of plants used by the korku tribe of Amravati District of Maharashtra, India. J Ethanopharmacol, 2006; 107(3): 463 469. 4. Jain A, Katewa SS, Chaudhary BL, Praveen G. Folk herbal medicines used in birth control and sexual diseases by tribals of southern Rajasthan, India. J Ethanopharmacol,2004; 90(1): 171 177. 5. Katewa SS, Chaudhary BL, Jain A. Folk herbal medicines from tribal area of Rajasthan, India. J Ethanopharmacol, 2004; 92(1): 41 46. 6. Sandhya B, Thomas S, Isabel W, Shenbagarathai R. Ehano medicinal plants used by the valaiyan community of Piranmalai Hills (reserved forest), Tamilnadu, India: A pilot study. Afri J Trad CAM, 2006; 3(1): 101 114. 7. Mc cann MC, Chen L, Roberts K, Kemsley EK, Sene C, Carpita NC, Stacey NJ, Wiloson RH. Infrared microspectroscopy: Sampling heterogeneity in plant cell wall composition and architecture. Physiol Planta, 1997; 100: 729 738. 8. Harborne JB. Phytochemical methods: A guide to modern thechniques of plant analysis, 3 rd edition, chapman and hall, 1998; UK. 9. Balunas MJ. Kinfhorn AD. Drug discovery from medicinal plant. Life Sci, 2005; 78: 431 441. Available online: www.ijipsr.com July Issue 1295
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