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Investigations on important secondary metabolites in Dolichandrone falcata Seem. Leaves Using GC-MS

Ekade P. P. Manik S. R.*
Department of Botany, Sant Gadge Baba Amravati University, Amravati – 444 602, Maharashtra, India
Corresponding Author: Dr. S. R. Manik, Professor & Head, Department of Botany Sant Gadge Baba Amravati University, Amravati – 444 602, Maharashtra, India Tel: +91-9850341510 E-mail: pravinekade005@gmail.com
Date of Submission: 22-04-2013 Date of Acceptance: 29-04-2013 Conflict of Interest: NIL Source of Support: NONE
Copyright: © 2013 Dr. S. R. Manik et al, publisher and licensee IYPF. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited.
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Abstract

Objective: To find out important secondary metabolites that gives much medicinal importance to Dolichandrone falcata in traditional tribal practices. Methods: The preliminary phytochemical screening is performed using the methods of Harborne and Raman. The preliminary screening reveals the presence of steroids, flavonoids, tannins and saponins. Gas Chromatography – Mass Spectroscopy analysis of Dolichandrone falcata leaves is performed on three extracts namely methanol, chloroform and dichloromethane solvents. Results: The methanol extract analysis shows the presence of many biologically important compounds like phytol, octadeconoic acid etc. The chloroform extract exhibit the presence of vitamin E, n-hexadecanoic acid etc. While dichloromethane extract analysis shows the highest amount of vitamin E and its derivatives. Conclusion: The compounds responsible for antimicrobial, anti inflammatory and anti-oxidant activity were found in the present analysis. Apart from this the leaves of Dolichandrone falcata is a good source of vitamin E and its derivatives.

Keywords:

Dolichandrone falcata, GC – MS, Phytochemical, Vitamin E.

Introduction

Dolichandrone falcata Seem. belonging to family Bignoniaceae is a small to medium size tree distributed in the central and southern parts of India[1]. The plant is used by the tribal of Northeast Maharashtra to cure stomach problems[2]. It is a traditional medicinal plant of Ayurveda used for the purpose of abortion and fish poisoning [3]. The plant leaves are reported to have anxiolytic effect on rats [4]. The bark paste of Dolichandrone falcata is applied in case of fractures [5]. The bark juice is used for menorrhagia and leucorrhoea[6]. The leaves of this plant are used as antioxidant, antiestrogenic and antidiabetic [7, 8]. Despite of these applications this plant is yet to be worked out for its chemical composition. The present study aims at the identification of maximum metabolites from Dolichandrone falcata leaves.

Material and Methods

`A. Collection of Plant Material: The leaves of Dolichandrone falcata were collected from Melghat forest of Amravati district, Maharashtra, India. The collected leaves were carefully examined for too old, etiolated, infected parts and were removed accordingly. Only fresh leaves were taken for the analysis. These leaves were dried in the shade till all its moisture gets evaporated. These dried leaves then converted to the powder form for further analysis.
B. Extraction: 10 gram of powder was extracted using Soxhlet apparatus for 24 hours in methanol. The extract then evaporated to dryness. At the time of analysis dried extract was dissolved in same solvent and this sample taken for GC – MS analysis. In the same way other two samples one in chloroform solvent and other in dichloromethane solvent were prepared. These three samples were then taken for GC – MS analysis.
C. GC – MS analysis: The analysis was carried out using gas chromatography – high resolution mass spectrophotometer. Dried extract were dissolved in the 5 ml of chloroform solvent. 2 μl of this solution was employed for GC – MS analysis. The GC-MS analysis was carried out using Alegent Hp 7880 with column of 30 meter length, 0.25 mm ID, 0.32 thickness. Helium gas was used as carrier gas at constant flow rate of 1ml/ minute. Injector temperature was set at 1000C. The oven temperature were programmed from 500C to 2800 C at 100 C/ minute to 2000C then100C/3 minutes to 2500C ending with a 5 minutes isothermal at 2800C. The sample was injected in split mode as 50:1. Identification of the compounds was done by comparing the spectral data of sample compound with the compound spectra present in spectral libraries (NIST).
Results
The extraction of Dolichandrone falcata leaves in three different solvents gives valuable information about its chemical constituents. The GC – MS data reveals the presence of eight compounds in methanol extract, nine compounds in chloroform extract and three compounds in dichloromethane extract. The compounds obtained with different solvent shows the 80% of metabolite different from each other. The most important metabolite identified in chloroform and dichloromethane extract is vitamin E, which shows its highest concentration in dichloromethane extract. β - tocopherol, the other derivative of vitamin E is found in dichloromethane extract. Identified metabolites and their relative concentrations are given in Table 1, 2 and 3.

Discussion

Dolichandrone falcata plant is known to be medicinal in traditional practices as well as proven by the latest research also. In the present study the attempts were made to find the chemical constituents of plant leaves to know its medicinal importance by using gas chromatography-mass spectroscopy approach. The results obtained in present study authenticate the earlier reports of therapeutic importance of the plant. The leaves are shown to have antioxidant property [7]. This property must be due to the presence of natural antioxidant vitamin E and its derivative β - tocopherol in leaves. Vitamin E found in chloroform extract with 52.57% concentration and in dichloromethane extract with 72.87% and its derivative tocopherol with 11.89% concentration. The leaves are shown to have the anti inflementary effect[7] the compound responsible for this property is n- hexadecanoic acid as this compound is known to have anti inflammatory properties[9]. n - hexadeconoic acid found in 24% and 9.24% in methanol and chloroform extract respectively. The antibacterial activity of longchain unsaturated fatty acids is well known. Fatty acids play the key role in antimicrobial food additives also[10], in present investigation we have find the octadeconoic acid i.e. stearic acid, which may be responsible for antimicrobial activity of plant leaves. Octadeconoic acid found in methanol with 3.08% concentration. Phytol which is found in methanol extract used as a precursor for the manufacture of synthetic forms of vitamin E[12], this compound may be used as precursor in natural biosynthesis of vitamin E in this plant. The analysis of Dolichandrone falcata definitely proves its potential as alternative medicinal plant in pharmaceuticals and drug formulations.

Acknowledgment

The authors express sincere thanks to Sophisticated Analytical Instrument Facility (SAIF), Indian Institute of Technology (IIT), Mumbai for supporting instrumental analysis for this research work.

Tables at a glance

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Table 1 Table 2 Table 3
 

Figures at a glance

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Figure 1 Figure 2 Figure 3
 

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