PHYTOCHEMICAL ANALYSIS OF ETHANOL EXTRACT OF AMPELOCISSUS LATIFOLIA (ROXB.) PLANCH TUBEROUS ROOT USING UV-VIS, FTIR AND GC-MS

PHYTOCHEMICAL ANALYSIS OF ETHANOL EXTRACT OF AMPELOCISSUS LATIFOLIA (ROXB.) PLANCH TUBEROUS ROOT USING UV-VIS, FTIR AND GC-MS

K. B. Theng * ¹ and A. N. Korpenwar ²

Department of Botany ¹, Shri Shivaji Science and Arts College Chikhli, Dist- Buldhana, M.S., India

Rashtrapita Mahatma Gandhi Science and Arts College Nagbhid ², Dist- Chandrapur, M. S., India

ABSTRACT: The present investigation was carried to determine UV-VIS, FTIR and GC-MS analysis of Ampelocissus latifolia tuberous root. The ethanol extract was examined under visible and UV light for the proximate analysis. The UV‐VIS profile of Ampelocissus latifolia tuberous root ethanol extract showed the peaks at 409 nm, 424 nm and 479 nm with the absorption of 1.682, 1.704 and 1.479 respectively. The FTIR spectrum was revealed the presence of alcohols, aromatic compound, alkanes, aldehydes, ketones, alkenes, amines, amides, nitro compounds, carboxylic acids, ethers, esters and alkyl halides in ethanol extracts of Ampelocissus latifolia tuberous root. GC-MS analysis of ethanol extract of Ampelocissus latifolia tuberous root was revealed the presence of 14 phytochemical compounds. The presence of various functional groups and phytocompounds in Ampelocissus latifolia tuberous root confirm that it act as a most important source of drugs against various ailments.

Phytochemical analysis, Ampelocissus latifolia,

UV-VIS spectrum, FTIR spectrum, GC-MS analysis

INTRODUCTION: Medicinal plants regarded as one of the main source of drug for the health of individual and communities ¹. The information about medicinal properties of plant has been obtained on the basis of various medicinal systems such as Ayurveda, Unani and Siddha during many centuries ². All the existing medicinal plants considered as main source for the invention of new drugs beneficial to human beings ³. Phytoconstituents are the natural bioactive compounds found within plants. These phytoconstituents along with nutrients form integral defense system to protect against various diseases ⁴.

During last few decades UV, FTIR and GC-MS were acted as powerful techniques for the identification, separation and structural determination of phytochemicals5. Phytoconstituents determination is easily performed by using gas chromatography (GC) and liquid chromatography (LC) technique with definite revealing system. Gas chromatography- mass spectroscopy (GC-MS) helped in identification of compounds at less than 1 mg 6. Commonly GC-MS applied for drug detection, environmental investigation and detection of unknown samples. GC-MS technique has been founds very effective for the separation and recognition of composite mixtures of phytochemicals 7.

Determination of chemical properties of phytocompounds present in plant provides necessary information about different functional groups revealing its medicinal properties 8. The FTIR analysis produced spectrum in which vibration of bond within chemical functional groups measure hence regarded as biochemical fingerprint of the sample 9. FTIR spectroscopy is known as time‐saving process to distinguish and identified functional groups 10.

Spectroscopic methods such as UV-Visible and FTIR can be used together or separately as conventional methods to detect phytoconstituents 11. Hence, the objective of the present study is to identify the phytochemical constituents of Ampelocissus latifolia by using UV-VIS, FTIR and GC-MS technique.

MATERIAL AND METHODS:
Collection of plant material:
The plant material of Ampelocissus latifolia was collected during September- October 2013 from Wasali forest area of Buldhana District, Maharashtra, India. Plant was identified by using various floras. Herbarium specimen of the plant was deposited at Department of Botany, Shri Shivaji Science and Arts College Chikhli. Tuberous roots were collected, thoroughly washed with water to remove foreign matter; shade dried and then grinds into fine powdered by using mechanical grinder.

Extraction of plant sample:
The grinded, fine powdered of tuberous root was subjected to extraction by using soxhlet apparatus. About 20 gm of tuberous root powdered was successively extracted with ethanol for 8 hrs. Ethanol extract was filtered through Whatman No. 1 filter paper and the filtrate was collected (crude extracts). Ethanol extract was concentrated, solidified and used for further studies.

UV-VIS Spectroscopic analysis:
Ethanol extract of Ampelocissus latifolia tuberous root was examined under UV and Visible light for immediate investigation. Such ethanol extract of tuberous root was centrifuged at 3000 rpm for 10 minutes and filtered through filter paper (Whatman No.1) under high pressure of vacuum pump. The sample was diluted to 1:10 by using same solvents. The extract scanned in the wavelength range from 190-1100 nm using EQUIPS- TRONICS (EQ-826) Spectrophotometer and the peaks were detected.

FTIR Spectroscopic analysis:
Dried powdered of ethanol extract of Ampelocissus latifolia tuberous root was used for FTIR analysis. The dried 10 mg of extract powdered was mixed with KBr salt and encapsulated in 100 mg of KBr pellet in order to prepare translucent sample discs. The infrared spectrum of solid was recorded in the scan range from 4400-450 cm-1 on a FTIR spectrophotometer, Perkin Elmer Spectrum (RX1) with a resolution of 1cm-1. Perkin Elmer spectrophotometer was used to detect characteristic peak and their functional group.

GC-MS analysis:
The composition of ethanol extract of Ampelocissus latifolia tuberous root was analyzed by using GC-MS instrument (AccuTOF GCV). The GC-MS system was equipped with a FID detector and capillary column of HP-1 (30 m x 0.25 mm; film thickness 0.25 μm). Helium was used as carrier gas at a flow rate of 1ml/min and a split injector (split ratio 1:10). The oven temperature was programmed from 80-2600 C at the rate of 10ºC/min and held at this temperature for 3 minutes and then increased to 280ºC at 5ºC/minute and held at this temperature for 9 min. The injector and detector temperatures were set at 2500 C and 2800 C respectively. Ethanol extract sample (0.1 μl) was injected into GC-MS instrument for its analysis. Ion source temperatures were maintained at 2000 C and the mass spectra were taken at 70eV.

Identification of Components:
Interpretation of mass spectrum GC-MS was conducted using data base of National Institute Standard and Technology (NIST) having more than 62,000 patterns. Spectrum of the unknown component was compared with the spectrum of known components stored in the NIST Library. The name, molecular weight, molecular formula and the structure of the components of Ampelocissus latifolia tuberous root were recorded.

RESULTS AND DISCUSSION:
The qualitative UV-VIS spectrum profile was selected for ethanol extract of Ampelocissus latifolia tuberous root powdered at a wavelength of 300 to 500 nm due to sharpness of the peak and proper baseline. The profile was shown peak at 409 nm, 424 nm and 479 nm with the absorption of 1.682, 1.704 and 1.479 respectively (Table 1).

TABLE 1: UV-VIS PEAK VALUES OF ETHANOL EXTRACT OF AMPELOCISSUS LATIFOLIA TUBEROUS ROOT

The UV-VIS spectrum profile of ethanol extract of Ampelocissus latifolia tuberous root was shown in Fig. 1.

FIG. 1: UV-VIS SPECTRUM OF ETHANOL EXTRACT OF AMPELOCISSUS LATIFOLIA TUBEROUS ROOT.

The FTIR spectrum was used to identify the functional group of different phytoconstituents based on the peak values in an infrared region of ethanol extract of Ampelocissus latifolia tuberous root powdered. The result of FTIR peak values were shown in Fig. 2.

The FTIR spectroscopic analysis of Ampelocissus latifolia was revealed the presence of alcohols, aromatic compound, alkanes, aldehydes, ketones, alkenes, amines, amides, nitro compounds, carboxylic acids, ethers, esters and alkyl halides in ethanol extracts of tuberous root powdered as shown in Table 2.

Pednekar and Raman, (2013) during FT-IR analysis of Ampelocissus latifolia (Roxb.) leaves was revealed that leaf powder showed alkynes, alcohols, esters, nitro compound, phenols, polysulfides and aliphatic iodo compounds. The stem powder showed alcohols, esters, aldehydes, ketones, phenols and nitro compound.

Raman et al., (2014) was reported the presence of alcohols, phenols, alkanes, aldehydes, alkenes, ethers, carboxylic acids, esters, alkyl halides, amines, amides, ketones, peroxides, etc. as functional groups during Fourier Transform Infrared (FT-IR) Spectroscopic study of Ampelocissus latifolia (Roxb.) Planch and it’s different leaf extracts.

FIG. 2: FTIR SPECTRUM OF ETHANOL EXTRACT OF AMPELOCISSUS LATIFOLIA TUBEROUS ROOT

Tab. 2: FTIR SPECTRUM OF ETHANOL EXTRACT OF AMPELOCISSUS LATIFOLIA TUBEROUS ROOT.

The GC-MS analysis of ethanol extract of Ampelocissus latifolia tuberous root was revealed the presence of 14 phytochemical compounds that could contribute to the medicinal property of the plant. The GC-MS chromatogram and mass spectrum of phytochemical compounds obtained in ethanol extract of Ampelocissus latifolia tuberous root were shown in Fig. 3 and 4.

A: GC-MS chromatogram of ethanolic extract of Ampelocissus latifolia tuberous root, A1: Mass spectra of Benzeneethanamine, 2, 5-difluoro- 3, 4-trihydroxy-N-methyl-, A2: Mass spectra of Pterin-6-carboxylic acid. A3: Mass spectra of I- Gala-I-ido-octose, A4: Mass spectra of Ethanol, 2-propoxy, A5: Mass spectra of Propane, 1-(1-methylethoxy)-, A6: Mass spectra of Propane, 2, 2- diethoxy-.

FIG. 3: GC-MS CHROMATOGRAM AND MASS SPECTRUMS OF AMPELOCISSUS LATIFOLIA TUBEROUS ROOT

A7: Mass spectra of Ethane, 1, 1, 1-triethoxy-, A8: Mass spectra of Dodecane, A9: Mass spectra of Azulene, 1, 2, 3, 4, 5, 6, 7, 8-octohydro-1, 4-dimethyl-7-(1-methylethenyl)-, [1S- (1 , 4 , 7 )]-,A10: Mass spectra of Naphthalene, 1, 2, 3, 4, 4a, 5, 6,8a- octahydro-7-methyl-4-methyethyl)-, (1 , 4a , 8a ) –, A11: Mass spectra of Naphthalene, 1, 2, 3, 4-tetrahydro-1, 6-dimethyl-4-(1-methylethyl)-, (1S-cis)-,A12: Mass spectra of Tetradecane, 2, 6, 10-trimethyl-, A13: Mass spectra of 1-Iodo-2-methylnonane, A14: Mass spectra of Nonadecane.

FIG.4: MASS SPECTRUMSOFPHY TO CHEMICAL COMPOUNDS OF AMPELOCISSUS LATIFOLIA TUBEROUS ROOT
The identification of phytochemical compounds were confirmed using data base of National Institute Standard and Technology (NIST) and their retention time (RT), molecular formula (MF), molecular weight (MW) and peak area in percentage (%)represented in Table 3. The first compound identified with less retention time (3.1min) was Benzeneethanamine, 2, 5-difluoro- 3, 4-trihydroxy-N-methyl- whereas Nonadecane was the last compound which took longest retention time (26.3 min). The major phytoconstituents present in ethanol extract of tuberous root were Benzeneethanamine, 2, 5-difluoro- 3, 4-trihydroxy-N-methyl-(31.21%), Pterin-6-carboxylic acid (31.21%), I- Gala-I-ido-octose(31.21%), Ethanol, 2-propoxy(0.53%), Propane, 1-(1-methylethoxy)- (0.53%), Propane, 2, 2- diethoxy-(0.84%), Ethane, 1, 1, 1-triethoxy-(0.84%), Dodecane(0.47%), Azulene, 1, 2, 3, 4, 5, 6, 7, 8-octohydro-1, 4-dimethyl-7-(1-methylethenyl)-,[1S- (1 , 4 , 7 )]-(0.18%), Naphthalene, 1, 2, 3, 4, 4a, 5, 6,8a- octahydro-7-methyl-4-methyethyl)-, (1 , 4a , 8a ) –(0.62%), Naphthalene, 1, 2, 3, 4-tetrahydro-1, 6-dimethyl-4-(1-methylethyl)-, (1S-cis)- (0.87%) , Tetradecane, 2, 6, 10-trimethyl-(0.48%), 1-Iodo-2-methylnonane(0.48%) and Nonadecane (0.48%) as listed in Table 3.

TABLE 3: GC-MS ANALYSIS OF AMPELOCISSUS LATIFOLIA TUBEROUS ROOT

Where, R.T. = Retention time, M.F. = Molecular formula, M.W. = Molecular weight.

CONCLUSION: UV spectrophotometric analysis is a simple, rapid and accurate method for the determination of phytoconstituents present in crude drug powdered of medicinal plants. The FTIR spectral analysis has shown the presence of characteristic functional groups in ethanol extracts of Ampelocissus latifolia plants. GC-MS analysis has revealed the presence of different active compounds of important medicinal value against various ailments in studied plants. Hence study revealed the isolation, characterization, purification and biological activity of specific active compounds are necessary for their further studies.

ACKNOWLEDGMENTS: We would like to thanks SAIF, IIT, Bombay, Powai, Mumbai, for providing the GC-MS facility. Authors are also thankful to SAIF, CIL, Panjab University, Chandigarh for providing FT-IR facilities and encouragement.

REFERENCES:
1. Antony Sandosh T, Paul John Peter M, Yesu Raj J. Phytochemical analysis of Stylosanthes fruticosa using UV-VIS, FTIR and GC-MS. Res. J. Chem. Sci. 2013; 3(11): 14-23.
2. Ragavendran P, Sophia D, Arul Raj C, Gopalakrishnan VK. Functional group analysis of various extracts of Aerva lanata (L.,) by FTIR spectrum. Pharmacologyonline. 2011; 1: 358-364.

3. Murugan M, Mohan VR. Phytochemical, FT-IR and antibacterial activity of whole plant extract of Aerva lanata (L.) Juss. Ex. Schult. Journal of Medicinal plants Studies. 2014; 2(3): 51-57.
4. Dipak K, Rupali S, Sayed I, Bhadange DG. Phytochemical screening of eight traditionally used ethnomedicinal plants from Akola district (ms) India. International Journal of Pharma and Bio Science. 2010; 1(4): 253-256.
5. Karpagasundari C, Kulothungan S. Analysis of bioactive compounds in Physalis minima leaves using GCMS, HPLC, UV-VIS and FTIR techniques. Journal of Pharmacognosy and Phytochemistry. 2014; 3(4): 196-201.
6. Sahu N, Saxena J. Phytochemical analysis of Bougainvillea Glabra Choisy by FTIR and UV-VIS spectroscopic analysis. Int. J. Pharm. Sci. Rev. Res. 2013; 21(1): 196-198.
7. Szarka S, Hethelyi E, Kuzovkina IN, Lemberkovics E, Eva S. GC-MS method development for the analyses of thiophenes from solvent extracts of Tagetes patula L Chromatographia.  2008; 68 (SUPPL. 1):S63-S69.
8. Ashokkumar R, Ramaswamy M. Phytochemical screening by FTIR spectroscopic analysis of leaf extracts of selected Indian Medicinal plants. International journal of Current Microbiology and applied Sciences. 2014; 3(1): 395-406.
9. Johnson M, Nazneen Bobby MD, Wesely EG. FT-IR studies on the leaves of Albizia lebbeck Benth. International Journal of Pharmacy and Pharmaceutical Sciences, 2012; 4(3): 293-296.
10. Grube M, Muter O, Strikauska S, Gavare M, Limane B. Application of FT-IR spectroscopy for control of the medium composition during the biodegradation of nitro aromatic compounds. Journal of Indian Microbiology and Biotechnology. 2008; 35: 1545–1549.
11. Ibrahim M, Hameed AJ, Jalbout A. Molecular Spectroscopic Study of River Nile Sediment in the Greater Cairo Region. Applied Spectroscopy, 2008; 62(3): 306-311.
12. Pednekar PA, Raman B. Antimicrobial and antioxidant potential with FTIR analysis of Ampelocissus latifolia (Roxb.) Planch. leaves. Asian Journal of Pharmaceutical and Clinical Research. 2013; 6 (1): 157-162.
13. Raman B, Pednekar PA, Kulkarni V. Study on Ampelocissus latifolia (Roxb.) Planch. and it’s different leaf extracts by Fourier Transform Infrared Spectroscopy. International Journal of Pharmacy and Pharmaceutical Sciences. 2014; 6 (7): 602-610

How to cite this article:

Theng KB and Korpenwar AN: Phytochemical Analysis of Ethanol Extract of Ampelocissus Latifolia (Roxb.) Planch Tuberous Root Using UV-Vis, FTIR and GC-MS. Int J Pharm Sci Res 2015; 6(9): 3936-42.doi: 10.13040/IJPSR.0975-8232.6(9).3936-42.

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