Chemical Characterization from GC-MS Studies of Ethanolic Extract of Macrotyloma uniflorum

 

V. Bharathi¹*, A. Vijaya Anand²

¹Ph.D. Scholar in Biochemistry, Bharathiyar University, Coimbatore, Tamilnadu

²Department of The Human Genetics and Molecular Biology, Bharathiyar University, Coimbatore, Tamilnadu

 

ABSTRACT:

Preliminary Phytochemical screening and GCMS analysis was performed on the seed and leaves formulation of Macrotyloma uniflorum (MUF) to find out the chemical constituents. Phytochemical screening revealed the presence of steroids, alkaloids, sugars, phenolics, flavonoids, saponins and tannins .The GCMS result revealed the presence of 38 different phytocompounds.viz.. Butyl cyclohexyl-acetat, tert-Butylcyclohexyl acetate, 1-Dodecanamine, N,N-dimet, phenol, 2,4-bis(1,1-dime, n,n-dimethyl-1-pentad, vundecanal, 2-methyl- ethanol, 2-(dodecyloxy, allyl-2,8,9-trioxa-5-a, Tetradecanoic acid, Hexadecanoic acid, Heptadecenoic acid, Ascorbic acid 2,6-dihexa,Pentadecenoic Acid, -Heptadecenoic acid, ethanol, 2-(dodecyloxy), Diethylene glycol monododecyl, Octadecanoic acid, methyl ester, 1,3,12-Nonadecatriene-5,Pentadecenoic acid, Octadecanoic acid, Methyl 17-methyl-octadecano hexatriacontane and 5,8,11,14-Eicosatetraenoic acid. The presence of various bioactive compounds confirms the application of MUF formulation for various diseases such as asthma, bronchitis , urinary discharges and for treatment of kidney stones. However, isolation of individual Phytochemical constituents may proceed to find a novel drug.

 

 

INTRODUCTION:

According to World Health Organization (WHO), more than 80% of the world’s population relies on traditional medicines for their primary health care needs (Duraipandiyan et. al ., 2006). Medicinal plants form the backbone of the traditional medicine in the last few decades.  Natural remedies from medicinal plants are found to be safe and effective. They are regarded as the potential sources of new compound of the therapeutic value and the source of lead compound.  Many plant species have been used in folkloric medicine to treat various ailments. Even today compounds from plants continue to play a major role in primary health care as therapeutic remedies in many developing countries (Gajalakshmi et al.,(2011).

 

If a single plant is used for various diseases it may contain more than one bioactive compound(s). If used in mixture they may show the synergic effect of two or more bioactive compounds. The use of different parts of plant for particular activity gives the idea that these bioactive compounds may be restricted to the particular part of plant body. Smolinski et al., (2003) Phytochemicals are secondary metabolic compounds found in plants. Many of these are known to provide protection against insect attacks and plant diseases, stimulation of the immune system, modulation of hormone metabolism and antibacterial and antiviral  effect (James et al.,1983).The most important of these bioactive constituents of plants are Alkaloids, Tannins, Flavonoids, Cardiac glycosides, Steroids and Saponins (Taylor, 2000). Macrotyloma uniflorum, commonly known as horse gram (Fabaceae) is a herbaceous plant with annual branches, sub erect or twining, leaflets 2.5-5 cm and widely distributed throughout Asia, Africa and Australia. It is prominent for its medicinal uses because different parts of the plants are used for the treatment of heart conditions, asthma, bronchitis, leucoderma, urinary discharges and for treatment of kidney stones. M. uniflorum could play a role in antioxidation . The plant is a common twining creeper, a branched sub-erect or trailing annual with small trifoliate leaves, bearing when mature, narrow flat curved pods 15-2.0 inches long tipped with a persistent style (Kottai Muthu et al., 2009). The pods contain 5-6 flattened, ellipsoid seeds 1/8-1/4 inch long. It distributes native to most parts of India, and is found up to altitude of 5000m. It was reported that seed (both tender and mature) contains are poor in amino acid content, but rich in urease. A new and nonspecific lectin having the inner carbohydrate moiety as N-acetyl glucosamine, N-M-glycosidically linked to aspargine has even isolated from seed of D. biflorus. Constituents of grain with husk: albuminoids starch, oil, fiber, ash, and phosphoric acid: enzyme urease (Nanta et al.,2011).

 

MATERIALS AND METHOD:

Collection of Plant Material:

The healthy plant samples of Macrotyloma uniflorum was collected from Trichy. The collected plant materials were transported to the laboratory. The plant materials were identified and authenticated at Department of Botony, St. Joseph's college, Trichy.

 

Preparation of Leaf Powder

The seed and leaves of Macrotyloma uniflorum  was collected, washed and cut into small pieces and dried at room temperature for two weeks and made in to powder for further analysis.

 

Extraction of Plant Material:

Anand, et al .,(2012) Aqueous, chloroform  and alcoholic extracts were prepared according to the methodology of Indian Pharmacopoeia. The shady dried plants materials were subjected to pulverization to get coarse powder. The coarse powder material was subjected to soxhlet extraction separately and successively with alcohol and distilled water. These extracts were concentrated to dryness in flash evaporator under reduced pressure and controlled temperature (40-50⁰C). The aqueous and alcohol extracts put in air tight containers stored in a refrigerator.

 

Phytochemical screening:

Evan et al., (2002), The MUF was tested for steroids, alkaloids, sugar, phenolic compounds, flavonoids, saponins, tannins, anthraquinone and amino acids. Phytochemical screening of the extract was carried out according to the standard method.

 

GCMS analysis:

Merlin  et al.,( 2009), The GCMS analysis of ethanolic crude extract of MUF  was performed using a GCMS equipment Thermo GC-TRACE ultra ver: 5.0, Thermo MS DSQ II. Experimental conditions of GCMS system were as follows: TR 5-MS capillary standard non-polar column, dimension: 30 m, ID: 0.25 mm, Film: 0.25 mm was used and flow rate of mobile phase (carrier gas: He) was set at 1.0 ml/min. In the gas chromatography part, temperature program (oven temperature) was 40⁰C raised to 250 ⁰C at 5 ⁰C/min and injection volume was   1 mL. Samples which dissolved in chloroform were run fully at a range of 50e 650m/z and the results were compared by using Wiley Spectral library search program. The mass spectra detected in 36 min.

 

RESULT AND DISCUSSION:

The Phytochemical screenings of MUF revealed that the ethanolic extract contains steroids, alkaloids, terpinoid, Phenol, Quinone and Protein. The Phenol and Quinone were absent in the Aqueous and chloroform extract. The ethanolic extract contain more phytoconstituent than compared to other extract (Table 1) .

 

Table 1.-

Extracts and tests	Extracts
	Aqueous	Ethanol	Chloroform
Alkaloids	+	+	+
Terpenoids	+	+	+
Phenols and Tannins	-	+	-
Sugar	-	-	-
Saponin	-	-	+
Flavonoids	-	+	-
Quinines	-	+	-
Proteins	+	+	+
Sterols	+	+	+

 

The GCMS analysis of  ethanolic extract of  MUF was presented  in the  Table 2. The result revealed the presence of 38 different phytocompounds.viz.. Butyl cyclohexyl-acetat, tert-Butylcyclohexyl acetate, 1-Dodecanamine, N,N-dimet, phenol, 2,4-bis(1,1-dime, n,n-dimethyl-1-pentad, vundecanal, 2-methyl- ethanol, 2-(dodecyloxy, allyl-2,8,9-trioxa-5-a, Tetradecanoic acid, Hexadecanoic acid, Heptadecenoic acid, Ascorbic acid 2,6-dihexa,Pentadecenoic Acid, -Heptadecenoic acid, ethanol, 2-(dodecyloxy), Diethylene glycol monododecy, Octadecanoic acid, methyl ester, 1,3,12-Nonadecatriene-5,Pentadecenoic Acid, Octadecanoic acid, Methyl 17-methyl-octadecano.Hexatriacontane and 5,8,11,14-Eicosatetraenoic acid. In terms of percentage of amount Hexadecanoic acid, Heptadecenoic acid, Pentadecenoic Acid were predominent in the extract. . N-Hexadecanoic acid– Heptadecenoic acid and Pentadecenoic Acid can be an antioxidant, hypocholesterolemic, nematicide, pesticide, lubricant  activities and hemolytic 5-alpha is a reductase  inhibitors.

 

 

CONCLUSION:

In the present study thirty eight chemical constituents have been identified from ethanolic extract of the MUF by Gas Chromatogram-Mass spectrometry (GC-MS) analysis. The presence of various Phytochemical contributes to the medicinal activity of the plant extract.

 

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Received on 30.12.2015          Modified on 10.01.2016

Accepted on 23.01.2016        © RJPT All right reserved