IDENTIFICATION OF LUPEOL IN ETHANOLIC EXTRACT OF CELASTRUS PANICULATUS BY HPTLC METHOD
HTML Full TextIDENTIFICATION OF LUPEOL IN ETHANOLIC EXTRACT OF CELASTRUS PANICULATUS BY HPTLC METHOD
Priyanka Sharma * 1, N. M. Shrivastava 2 and Savita Shrivastava 3
Department of Pharmacy, Barkatullah University 1, Bhopal - 462026, Madhya Pradesh, India.
Gandhi Medical College 2, Bhopal - 462001, Madhya Pradesh, India.
Government Maharani Laxmi Bai Girls Post Graduate (Autonomous) College 3, Bhopal - 462001, Madhya Pradesh India.
ABSTRACT: Celastrus paniculatus is a known herb used in alternative system of medicines since ancient times. Traditionally seed oil of Celastrus paniculatus has been in various medicinal remedies to cure neuronal disorders, asthma, cough, etc. The constituents found in Celastrus paniculatus reportedly are sesquiterpenoids, triglycerides, triterpenoids. A high-performance thin layer chromatography, which is considered as a sensitive method for accurate quantification method for analysis of compounds. The instrument CAMAG Linomat 5 lamp was used, and the wavelength was 254 nm.
Keywords: |
Celastrus paniculatus, Seed, Seed oil, Ethanolic extract, HPTLC, Lupeol
INTRODUCTION: Celastrus paniculatus commonly known as Jyotishmati, Intellect-tree, Climbing staff tree, Malkangani, Black oil tree. It is an important medicinal plant used in traditionally in Ayurvedic system of medicines it grows throughout India height of almost 1800-2000 meters. It is a deciduous wine which grows very large. This plant belongs to the class Angiospermae, order Celastrales and family Celastraceae. In the ancient time mostly seed oil of Celastrus paniculatus have been used for many remedies, but other parts of the plant are also very useful. Various part of the plant is used to treat malaria, oil is used as stomachic, in treatment of headache, leucoderma.
The reported chemical constituents of celastrus paniculatus are Malkangunin, celapanin celapanigin, celapagin, pristemerin, zeylasterone and zeylasteral, fatty oil with palmitic, oleic, linoleic and linolenic acids. A triterpene compound Lupeol was isolated from pet ether extract of the leaves which have wound healing activity 1-6. However, till now, all the chief chemical constituents have been reported in the seed, and its oil and other compounds have been reported in pet ether extract, methanolic and aqueous extract in another part of the plant. So in this paper, we are the discussing the presence of lupeol in the ethanolic leaves extract of the plant by HPTLC method 7-9.
MATERIALS AND METHODS: The fresh leaves of the plant were collected from district Vidisha in the month of July-August and authentication of the plant was done by the MFP PARC (Vindhya Herbals). Analytical grade solvents were used for the process. Lupeol standard was procured from Sigma Aldrich. The whole HPTLC estimation was performed in the Centre of Excellence in Biotechnology (MPCST, Bhopal)
Preparation of Extract: The leaves were shade dried and ground in the mixer. The coarse powder of leaves was further macerated for extractive values, and then 95 percent ethanol was selected for preparation of the extract.
High-Performance Thin-Layer Chromatography:
Instrument: The instrument CAMAG Linomat 5 lamp was used, and the wavelength was 254 nm and CAMAG TLC Scanner. The chromatographic separation was performed The HPTLC method was performed by using silica gel 60 F 254 plates (E. Merck KgaA 20.0 cm × 10.0 cm ) were used as a stationary phase to confirm and solvent system Toluene: Chloroform: Methanol (4:47:1.3)(v/v/v) were used. The Rf value was 0.94 and software win CATS Planar used. Other specifications are:
Number of Tracks | 9 |
Position of first track X | 15.0 mm |
Scan start pos. Y | 5.0 mm |
Distance between tracks | 21.2 mm |
Scan start pos. Y | 5.0 mm |
Slit dimensions | 6.00×0.40 mm, Macro |
Optimize optical system | Light |
Scanning speed | 20 mm/s |
Data reduction | 100 µm/step |
Measurable Table
Wavelength | 254 |
Lamp | D2 & W |
Measurement Type | 5.0 mm |
Measurement Mode | Remission |
Optical filter | Second order |
Detector mode | Automatic |
PM high voltage | 295 V |
RESULTS AND DISCUSSION: By analyzing the sample and standard, it was found that the standard Lupeol has Rf value 0.94. Regression via height: Single level Y = 0+0* X r =0.00000 Std dev = -1 was observed. The spectrum that is obtained is that of the test sample shown in Fig. 2 is super-imposable with standard Lupeol shown in Fig. 3, and this indicate the purity of the peak.
TABLE 1: TEST SAMPLE PEAK
Peak | Start
Rf |
Start Height | Max
Rf |
Max Height | Max
% |
End
Rf |
End height | Area | Area
% |
Assigned Substance |
1 | 0.93 | 0.6 | 0.94 | 47.0 | 100.00 | 0.97 | 0.3 | 505.5 | 100.00 | Substance 1 |
TABLE 2: STANDARD PEAK
Peak | Start
Rf |
Start Height | Max
Rf |
Max Height | Max
% |
End
Rf |
End height | Area | Area
% |
Assigned Substance |
1 | 0.92 | 2.7 | 0.94 | 46.9 | 100.00 | 0.97 | 1.0 | 531.6 | 100.00 | Substance 1 |
CONCLUSION: By analyzing the whole HPTLC profile the conclusions that have been made that the apart from seed and seed oil other parts of the plant also contains many constituents and especially ethanolic extract of the plant contains Lupeol, and by this result, there is high possibilities this may contain other important constituents.
ACKNOWLEDGEMENT: We are very grateful to Dr. R. K. Garg (Scientific Officer) and Dr. Anita Tilwari (Scientific Officer) of Centre of Excellence in Biotechnology, M.P. Council of Science and Technology, Bhopal for providing their kind support for HPTLC analysis in their esteemed lab.
CONFLICT OF INTEREST: Nil
REFERENCES:
- The Siddha Pharmacopoeia of India 1: 207-209.
- Kirtikar KR and Basu BD: Indian Medicinal Plants; Vol 1. Lalit Mohan Basu,Allahabad India 1984: 677-80.
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- Harish BG, Krishna V, Kumar HS, Ahamed BMK, Sharath R and Swami HMK: Wound healing activity and docking of glycogen-synthase-kinase-3-beta-protein with isolated triterpenoid lupeol in rats. Phytomed 2008; 15(9): 763-67.
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How to cite this article:
Sharma P, Shrivastava NM and Shrivastava S: Identification of lupeol in ethanolic extract of Celastrus paniculatus by HPTLC method. Int J Pharm Sci & Res 2014; 5(9): 3876-78. doi: 10.13040/IJPSR.0975-8232.5(9).3876-78.
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Article Information
42
3876-3878
617
1318
English
IJPSR
P. Sharma *, N. M. Shrivastava and S. Shrivastava
Department of Pharmacy, Barkatullah University, Bhopal, Madhya Pradesh, India.
sharma_priyanka30@yahoo.co.in
10 March 2014
25 April 2014
20 June 2014
10.13040/IJPSR.0975-8232.5(9).3876-78
01 September 2014