Int J Pharm Pharm Sci, Vol 7, Issue 6, 423-425Short Communication


GC-MS ANALYSES OF LEAF AND ROOT EXTRACTS OF DIDYMOCARPUS TOMENTOSA

J PRAMEELA1*, H RAMAKRISHNAIAH1, V KRISHNA2, A P DEEPALAKSHMI1

1Department of P G Studies and Research in Biotechnology, Government Science College, Bangalore, India, 2P G Department of Studies and Research in Biotechnology and Bioinformatics, School of Biological Sciences, Kuvempu University, Karnataka, India
Email: hramabt@yahoo.com

Received: 09 Mar 2015 Revised and Accepted: 02 May 2015

ABSTRACT

Objective: Didymocarpus tomentosa Wight., is a traditional medicinal plant used in the treatment of fever and skin allergy and the present study were conducted to identify the phytochemical constituents in leaf and root extracts using gas chromatography and mass spectrometry (GC-MS).

Methods: The methanolic leaf and root extracts were analyzed using Shimadzu GCMS-QP 2010 gas chromatography-mass spectrometer. The mass spectrum of GC-MS was interpreted using the database of the National Institute of Standards and Technology (NIST).

Results: The GC-MS analyses of leaf and root extracts revealed the presence of twenty six and twenty three phytochemical constituents respectively. 5-Hydrxoymethylfurfural (15.1%), cis, cis, cis-7,10,13-Hexadecatrienal (12.5%), Hexadecanoic acid (11.8%), Lupenone (29.1%), γ-Sitosterol (21.3%) and Lupeol (12.0%) were the major constituents.

Conclusion: The leaf and root extracts of D. tomentosa possess various phytochemical constituents, which are of high therapeutic values.

Keywords: Didymocarpus tomentosa, 5-Hydrxoymethylfurfural, Lupenone, Lupeol.

For thousands of years traditional herbal medicines have cured many diseases due to their special healing efficacy. It has gradually attracted interest and acceptance by the general public as efficient and cost effective medicine besides being locally available and safe. As a result, in recent years, there is an increasing trend to determine phytochemical composition of such traditional plants and GC-MS has become a powerful analytical tool for efficient separation of chemical constituents in the field of herbal medicines [1]. The phytochemical profiling of the extracts using GC-MS provides reliable data for the identification of medicinal useful compounds.

Didymocarpus of Gesneriaceae, comprises of 70 species [2]. Only a few species are chemically studied and most of these species are found distributed in India. Didymocarpus tomentosa Wight., [Syn. of Henckelia incana (Vahl) Spreng.] endemic to the Peninsular India, is traditionally used in the treatment of fever and skin allergy [3, 4]. Their extracts possess antioxidant and anti-inflammatory activities [5]. However, photochemical constituents of their extracts have not been reported till date. Therefore, in the present study, we report for the first time the GC-MS fingerprinting of leaf and root extracts which was carried out so as to establish volatile chemical constituents of D. tomentosa and to identify medicinally useful compounds.

Plant materials of Didymocarpus tomentosa were collected in the rocky hills of Savandurga, about 50 km west of Bangalore. The plant was identified using the Flora of British India and the Flora of the Tamil Nadu Carnatic [6, 7]. It was further authenticated by the National Ayurveda Dietetics Research Institute, Bangalore. A voucher specimen has been deposited at the above Centre (RRCBI/MCW/09). The collected plant materials were washed with tap water. They were air-dried thoroughly under shade at room temperature for 15-20 days. The shade dried materials were powdered using a mixer and stored in air‐tight container.

Leaf and root powder were extracted separately with methanol using hot extraction method. A total of 50 g of each powder was extracted with 500 ml methanol (1:10) in a Soxhlet apparatus for 2 days. The residue was removed by filtration through What Mann No. 1 filter paper and the extracts were concentrated using a Rotary evaporator. The extracts thus obtained were subjected to GC-MS analyses.

GC-MS analysis of leaf and root extracts were carried out using Shimadzu GCMS-QP 2010 gas chromatography-mass spectrometer interfaced with a turbo mass quadrupole mass spectrometer, fitted with an Rtx-5 fused silica capillary column (30 m × 0.25 mm, with 1 μm film thickness); the oven temperature was programmed from 150 °C to 210 °C (10 min) at 2 °C/min and increased to 260 °C (10 min) at 10 °C/min. Helium was used as a carrier gas at a flow rate of 1.0 ml/min; the injector temperature was 210 °C, injection volume of 1 μl and with a split ratio of 1: 200. The interface and MS ion source were maintained at 280 °C and 250 °C respectively, the mass spectra were taken at 70 eV with a mass scan range of 40-500 amu. Data handling was done using GC-MS solution software. The mass spectrum of GC-MS was interpreted using the database of NIST.

The GC-MS analysis of leaf extract resulted in the identification of twenty six constituents representing 100% of the chromatographical leaf extract. The phytoconstituents of the leaf extracts with their peak number, retention time (RT), name, molecular formula, molecular mass (MW) and area percentage (%) is presented in table 1.

Table 1: The phytoconstituents in the leaf extract of D. tomentosa

Peak no. RT Name of the phytoconstituents Mol. formula MW %
1 4.6 5-Methylfuran-2-aldehyde C6H6O2 110 0.8
2 4.8 2, 4-Dihydroxy-2, 5-dimethyl-3 (2H)-furan-3-one C6H8O4 144 0.9
3 5.6 5,6-Dimethyl-2,3-dihydro-1,4-dioxin C6H10O2 114 0.9
4 5.7 4-methyl-2-prop-1-enyl-1,3-dioxolane C7H12O2 128 1.2
5 5.9 2, 5-Dimethyl-4-hydroxy-3 (2H)-furanone C6H8O3 128 0.6
6 6.2 2-Hydroxy-3-methyl-4-pyrone C6H6O3 126 4.0
7 6.7 2, 5-Monomethylene-1-rhamnitol C7H14O5 178 0.9
8 7.2 2,3-Dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one C6H8O4 144 2.3
9 7.9 1, 2-Benzenediol C6H6O2 110 2.3
10 8.4 5-Hydrxoymethylfurfural C6H6O3 126 15.1
11 8.6 1, 2, 3-Propanetriol monoacetate C5H10O4 134 1.3
12 9.6 2-Methoxy-4-vinyl phenol C9H10O2 150 3.8
13 11.2 2-Hydroxy-6-methylbenzaldehyde C8H8O2 136 4.6
14 14.0 2-Amino-3-(3,4-dihydroxy-phenyl)-propionic acid C9H11NO4 197 4.3
15 14.2 (E)-13-Docosenoic acid C22H42O2 338 0.7
16 15.8 3,7,11,15-Tetramethyl-2-hexadecen-1-ol C20H40O 296 1.1
17 16.6 Methyl hexadecanoate C17H34O2 270 1.0
18 17.0 Hexadecanoic acid C16H32O2 256 11.8
19 18.6 Methyl 3-hydroxyoctadecanoate C19H38O3 314 1.0
20 18.8 cis,cis,cis-7,10,13-Hexadecatrienal C16H26O 234 12.5
21 20.4 Octadecanal C18H36O 268 4.3
22 21.9 Palmitic acid 2-hydroxy-1-(hydroxymethyl) ethyl ester C19H38O4 330 5.3
23 23.3 Propylene glycol monoleate C21H40O3 340 5.2
24 23.4 Methyl (Z)-5,11,14,17-eicosatetraenoate C21H34O2 318 4.8
25 27.4 Stigmasta-5, 22-dien-3-ol C29H48O 412 0.8
26 27.8 γ-Sitosterol C29H50O 414 8.6

The GC-MS analysis of root extract revealed the presence of twenty three constituents representing 100% of the chromatographical root extract. The phytoconstituents of the root extract is presented in table 2.

Table 2: The phytoconstituents in the root extract of D. tomentosa

Peak no. RT Name of the phytoconstituents Mol. formula MW %
1 6.2 1,3,5-Triazine-2,4,6-triamine C3 H6 N6 126 0.5
2 7.2 2,3-Dihydro-3,5-dihydroxy-6-methyl-4 (H)-pyran-4-one C6H8O4 144 0.4
3 8.3 5-Hydrxoymethylfurfural C6H6O3 126 0.6
4 9.6 2-Methoxy-4-vinyl phenol C9H10O2 150 0.4
5 11.2 2-Hydroxy-6-methylbenzaldehyde C8H8O2 136 0.9
6 16.6 Methyl hexadecanoate C17H34O2 270 0.8
7 17.0 Hexadecanoic acid C16H32O2 256 4.0
8 17.7 9-Octadecenoic acid C18 H34 O2 282 0.3
9 18.3 Cis, cis-octadeca-9,12-dienoic acid methyl ester C19 H34 O2 294 3.2
10 22.1 4H-1-Benzopyran-4-one, 2,3-dihydro-5,7-dimethoxy-2-phenyl- C17H16O4 284 2.8
11 22.6 (2S)-5-Methoxy-6-methyl-2-phenyl-7-chromanol C17H18O3 270 0.4
12 22.9 4-Phosphalotricyclo[6.1.1.0(2,6)]dec-2-ene, 4,9,9-trimethyl- C12 H19 P 194 3.0
13 23.1 1-Benzylidene-2-(diphenylmethylene) hydrazine C20H16N2 284 3.4
14 23.8 4-Nitrophenylsuccinic acid C10H9NO6 239 3.8
15 24.2 2,6,10,15,19,23-Hexamethyl-2,6,10,14,18,22-tetracosahexaene C30H50 410 0.6
16 25.9 2,7,8-Trimethyl-2-[(4R,8R)-4,8,12-trimethyltridecyl]-6-chromanol C28H48O2 416 0.4
17 26.2 Cholesta-4, 6-dien-3-ol C27H44O 384 0.4
18 27.2 Ergost-5-en-3β-ol C28 H48 O 400 0.9
19 27.4 Stigmasta-5,22-dien-3-ol C29 H48 O 412 2.3
20 27.8 γ-Sitosterol C29 H50 O 414 21.3
21 28.3 Lupenone C30H48O 424 29.1
22 28.4 Lupeol C30H50O 426 12.0
23 28.7 4-Stigmasten-3-one C29H48O 412 8.6

The majority of the phytoconstituents identified in the extracts is attributed with various biological activities. For example, Hydroxymethylfurfural (5-Hydroxymethylfurfural), a known cytotoxic phytochemical may prevent cancer-related processes in breast, prostate and endometrial cancer cells [8]. It also possesses nematicidal activity [9], antioxidant and anti-inflammatory activities [10] and antiproliferative activities [11]. Another major constituent present in both the extracts was γ-Sitosterol, which is known for lowering the cholesterol in human beings [12]. Recently, it has also been reported as an antidiabetic agent [13]. Hexadecanoic acid a very common saturated fatty acid is known as an anti-inflammatory phytoconstituent as it is a phospholipase inhibitor [14] and it's also known for its antibacterial activity [15]. Recently, Lupenone and Lupeol have been reported to inhibit protein tyrosine phosphatase 1B (PTP 1B) a possible alternative to the type 2 diabetes and obesity drug development [16]. In addition, Lupeol is attributed with anticancer activity [17, 18], anti-inflammatory activity [19], antiarthritic action [20, 21] and antifungal properties [22]. Lupenone has more recently been used in the treatment for hypopigmentation diseases [23]. Therefore the presence of various bioactive compounds in the leaf and root extracts of D. tomentosa justifies the use of the plant in the treatment of different ailments by the traditional practitioners and it further holds promise for the production of novel pharmaceuticals.

CONFLICT OF INTERESTS

The authors declare that there is no conflict of interest.

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