GC-MS Analysis of Some Bioactive Constituents of Diospyros virginiana

 

S.Priya¹*, S. Nethaji¹ and B. Sindhuja²

¹P.G & Research Department of Biochemistry, Marudupandiyar College,

Vallam, Thanjavur-613403, Tamil Nadu, India

²P.G Department of Biotechnology, S.T.E.T. Women’s College, Mannargudi, Tamil Nadu, India

 

ABSTRACT:

The present study of phytochemical analysis in the leaves and bark powder extract with absolute ethanol, the phytochemical compound screened by GC-MS method. In this GC-MS analysis, 40 bioactive phytochemical compounds and 30 compounds were identified in leaves and bark of Diospyros virginiana. The identified compounds predominantly phenolic derivatives are present included Hydrocarbons, Carbohydrates, Fatty Acid, Fatty Acid ester, Alcoholic compounds, Alkaloids, Ketones and Alkenes compounds. These different active phytochemicals have been found to possess a wide range of activities, which may help in the protection against incurable diseases.

 

 

INTRODUCTION:

Today natural products derived from plants are being tested for presence of new drugs with new modes of pharmacological action. A special feature of higher plants is their capacity to produce a large number of secondary metabolites¹. Recent studies are involved in the identification and isolation of new therapeutic compounds of medicinal importance from higher plants for specific diseases^{2, 3}. Knowledge of the chemical constituents of plant is helpful in the discovery of therapeutic agent as well as new sources of economic materials like oil and gums. The most important bioactive constituents of the plants are alkaloids, tannins, flavonoids and phenolic compounds. In India large number of plant species had been screened for their pharmacological properties but still a vast wealth of endangered species are unexplored. Medicinal plants are at interest to the field of biotechnology, as most of the drug industries depend in part on plants for the production of pharmaceutical compounds.

 

Diospyros virginiana is a persimmon species commonly called the American Persimmon,  Common Persimmon,  Eastern Persimmon,  "'Simmon", "Possumwood", or  "Sugar-plum".  This is a well-known indigenous tree, growing in woods and fields. Persimmons have been used to lubricate the lungs and strengthen the spleen and pancreas ⁴.

 

 

They improve energy and contain enzymes that help damaged cells and foreign microbes be broken down. Persimmons have a special affinity for the large intestines and heart. Persimmons have been used to treat bronchitis, catarrh, cough, diarrhea, dysentery, goiter, hangover, hemorrhoids and hiccoughs⁵. The bark has been used in intermittent and both it and the unripe fruit have been beneficial in various forms of disease of the bowels, chronic dysentery, and uterine hemorrhage; used in infusion, syrup, or vinous tincture⁶.

 

Seeds and fruits are generally low in crude protein, crude fat, and calcium but high in nitrogen-free extract and tannin^7. The inner bark and unripe fruit are sometimes used in treatment of fevers, diarrhea, and hemorrhage. Indelible ink is made from fruit. Persimmon is valued as an ornamental because of its hardiness, adaptability to a wide range of soils and climates, its lustrous leaves, its abundant crop of fruits, and its immunity from disease and insects. Hence the objective of the present study is to identify the phytochemical constituents with the aid of GC-MS technique.

 

MATERIALS AND METHODS:

Collection of plant material:

D.virginiana belongs to the family Ebenaceae was collected from Coonoor, Nilgiris District, Tamil Nadu, India and identified by the special key given Cambell flora⁸. The leaf and bark of D.virginiana were washed with sterile distilled water. After, the leaves and bark were shade dried and powdered by using pestle and mortar. 25g of powder was filled in the thimble and extracted successively with ethanol using a Soxhlet extractor for 48 h. The extracts were concentrated using rotary flash evaporator and preserved at 5°C in airtight bottle until further use. The ethanolic extracts of the plant was used for GC-MS analysis.

 

GC –MS analysis:

Preparation of extract:

Each 2 μl of the ethanolic extract both leaf and bark of D. virginiana was employed for GC/MS analysis.

 

Instruments and chromatographic conditions:

GC-MS analysis was carried out on a GC clarus 500 Perkin Elmer system comprising a AOC-20i autosampler and gas chromatograph interfaced to a mass spectrometer (GC-MS) instrument employing the following conditions: column Elite-1 fused silica capillary column (30 × 0.25 mm ID ×1 EM df, composed of 100% Dimethyl poly siloxane), operating in electron impact mode at 70 eV; helium (99.999%) was used as carrier gas at a constant flow of was employed (split ratio of 10:1) injector temperature 250°C; ion-source temperature 280°C. The oven temperature was programmed from 110°C (isothermal for 2min), with an increase of 10°C/min, to 200°C/min, then 5°C/min to 280°C/min, ending with a 9 min isothermal at 280°C. Mass spectra were taken at 70 eV; a scan interval of 0.5 s and fragments from 40 to 550 Da.

 

Identification of components:

Interpretation on mass spectrum of GC-MS was done using the database of 1ml/min and an injection volume of 0.5 EI having more than 62,000 patterns. The mass spectrum of the unknown component was compared with the spectrum of the known components stored in the WILEY library. The name, molecular weight and structure of the components of the test materials were ascertained.

 

RESULT AND DISCUSSION:

GC-MS chromatogram of the ethanolic extract of D. virginiana leaf (Figure 1) showed 27 peaks and bark (Figure 2)  reported 21 peaks indicating the presence of phytochemical constituents. On comparison of the mass spectra of the constituents with the WILEY library the phytoconstituents were characterized and identified    (Table- 1).

 

 

Fig 1: GC-MS Chromatogram of ethanolic leaf extract of Diospyros virginiana

 

Fig 2 GC-MS Chromatogram of ethanolic bark extract of Diospyros virginiana

 

From the results, it was observed that Hexadecanoic acid (synonym: Palmitic acid), 3, 7, 11, 15- Tetramethyl-2-hexadecen-1-ol (Synonym: Phytol), were the major components in the extract. Palmitic acid is reported to be an antioxidant, a nematicide and a pesticide ⁹ while Phytol are said to be cancer preventive. Other antioxidants present were Tetradecanoic acid; Hexadecanoic acid; Heptadecanoic acid; 2, 6, 10, 15, 19, 23– hexamethyl- (synonym: Squalene); gamma-Tocopherol.

 

The source of many plants (herbs and spices) can often be identified from the peak pattern of the chromatograms obtained directly from headspace analysis. Similarly, unique qualitative and quantitative patterns from a GC analysis will often help identify the source of many alcoholic beverages. The technique of fingerprint could really identify the false herbal products. The construction of chromatographic fingerprints aims at evaluating the quality of Herbal Medicines ¹⁰.

 

In the present study chemical constituents have been identified from ethanolic extract of the leaf and bark of Diospyros virginiana by Gas Chromatogram- Mass spectrometry (GC-MS) analysis. The presence of various bioactive compounds justifies the use of the whole plant for various ailments by traditional practitioners. However isolation of individual phytochemical constituents and subjecting it to biological activity will definitely give fruitful results. The present study, which reveals the presence of components in Diospyros virginiana suggest that the contribution of these compounds on the pharmacological activity should be evaluated.

 

Table-1 Phytocomponents identified in the ethanolic leaf and bark extracts of D.virginiana by GC-MS

No	RT		Name of the compound	Plant part	Molecular formula	MW	Peak Area
	Leaf	Bark
1 2   3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22   23 24   25 26 27 28 29 30 31 32	3.96 4.82   5.00 5.93 6.67 6.98 7.56 8.30 8.95 9.43 9.87 10.04 10.98 11.02 11.25 11.32 12.07 13.00 13.84 13.93 13.97 14.85   15.00 16.18   16.25 16.64 23.22	2.32 4.82   5.93 13.34   22.03 20.00 8.95 19.03 19.57   11.86         16.04   21.55   19.01           23.02 22.65   6.67 14.85 15.28 20.05 20.09 20.14	Undecanoic acid 4H-pyran-4-one,2,3-dihydro-3,5-dihydroxy- 6-methyl- 1,2 Benzene dicarboxylic acid Hexadecanoic acid, methyl ester Eicosane Glycerol or 2-phospho glyceric acid Oleic acid Palmitate Valeric acid Heptonic acid Lauric acid or n-do decanoic acid Phthalic acid Tetra decanoic acid or myristic acid Borazine Carbonic acid Acrylic acid ethyl ester Silane Hexadecanoic acid, ethyl ester 9,12 octadecanoic acid (Z,Z) 9-octa decanoic acid (Z) methyl ester Phytol 4-H-Pyran-4-one,2,3-dihydro-3,5-dihydroxy-6-methyl 1,2,3-Benzenetriol Naphthalene,dehydro-2-methoxy- 3-0-methyl-α-glucose Linoleic acid ethyl ester Α-D-glucopyranoside,o-α-D-glucopyranose gamma-Tocopherol 2,3-Anhydro-d-galactosan Margarinic acid, N-heptadecanoic acid Squalene Eicosanoic acid	Leaf, bark Leaf   Leaf, bark Leaf, bark Leaf Leaf, bark Leaf, bark Leaf, bark Leaf, bark Leaf, bark Leaf Leaf, bark Leaf Leaf Leaf Leaf Leaf, bark Leaf Leaf, bark Leaf Leaf, bark Leaf   Leaf Leaf, bark   Leaf, bark Leaf, bark Leaf Bark Bark Bark Bark Bark	C11H22O2 C6H8O4   C24H38O4 C17H34O2 C20H40 C3H6O4 C18H34O2 C16H32O2 C5H10O2 C7H114O2 C12H24O2 C6H4(COOH)2 C14H28O2 B3H6N3 H2Co3 CH2=CHCO2H H4Si C18H3602 C18H2202 C11H3602 C20H40O C6H8O4   C6H6O3 C11H20O   C7H14O6 C20H36O2 C6H6O3 C28H48O2 C6H8O4 C17H34O2 C30H50 C20H40	186 144   390 270 280 106 282 256 102 130 200 166 228 80 62 72 32 284 280 296 296 144   126 168   194 308 416 144 114 410 280	2.37 0.51   4.97 1.07 1.47 0.49 0.63 7.70 1.83 0.40 7.20 1.99 2.68 32.87 8.30 0.49 2.86 0.63 0.31 0.83 1.64 6.18   3.27 10.58   7.51 5.37 4.50 0.43 1.27 0.57 2.92 0.33

CONCLUSION:

The plant leaves and bark of Diospyros virginiana screened for bioactive compounds seemed to have the potential to act as a source of useful drugs and also to improve the health status of the consumers as a result of the presence of various compounds that are vital for good health.

 

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Received on 13.01.2014       Modified on 17.02.2014

Accepted on 21.02.2014      © RJPT All right reserved