INTRODUCTION:

Morina longifolia Wall. of family Dipsaceae is found at the height of 3000-3500 m. It has spiny margined leaves and long interrupted spike of flowers, but flowers white to rose pink and paired bracts subtending whorls with an enlarged ovate base and fused below. Flowers with long slender corolla tube to 2.5 cm, somewhat two lipped the lips to 6 mm, hairy. Leaves strap-shaped with shallow 3-spined lobes and long pointed spiny apex (Polunin and Stainton, 1984).Itis commonly known as "Whorl flower". Its stem leaves and flowers are used in Tibetan medicine. They are said to have a sweet and astringent taste with a heating potency. They are digestive, emetic and stomachic and are used in the treatment of stomach disorders (Tsarong, 1994). The plant possesses strong aromatic properties, used as incense and in the preparation of dhup, agarbatties, etc (Chopra 1996). The root paste has been applied externally on wounds and the aroma of the flowers has been used for unconsciousness in Indian traditional medicine (Gaur, 1999).The plant used in treatment of maggot wounds (Handoo, 2006).

Previous reports on Morina species showed that five new phenylpropanol derivatives, called morinins, as well as two known compounds as 3, 4-dimethoxycinnamylalcohol methyl ether, and p-methoxycinnamaldehyde, reported from the methanol extracts of the roots of the medicinal Chinese plant, M. chinensis.( Bao-Ning SU et al., 1999)

A novel acylated flavonol glycoside, quercetin 3-O-[2' "-O-(E)-caffeoyl]-alpha-L-arabinopyranosyl-(1-6)-beta-D-galactopyranoside was isolated from whole plant of Morina nepalensis var. alba (Tang et al., 2002). Seven new phenylpropanol derivatives, named morinins A-G (1-7), along with five known compounds, 4-O-methylcinnamyl alcohol, 4-O-methylcinnamyl methyl ether, 4-O-methylcinnamyl acetate, p-methoxybenzaldehyde, and 4-O-methyl-(E)-coniferyl alcohol, have been isolated from the roots of the medicinal Chinese plant, Morina chinensis (Yoshihisa Takaishi et al., 1999). Two novel tetrahydropyran sesquineolignans with a new carbon skeleton, named morinols A-B and other ten novel neolignans, named morinols C-L along with two known lignans, pinoresinol and lariciresinol, have been isolated from the roots of Chinese medicinal herb, Morina chinensis (Bao-Ning Su et al., 1999). In general, wild plants have been regarded as a natural reservoir of novel and more exotic fragrances however even after the assessment of Morina longifolia properties and its use in traditional medicine; the attention has been limited because of the lack of information about its chemical composition (S.K. Bhattacharjee, 2000)

Volatile oils are generally complex mixtures of different compounds like sesquiterpenes. Potential synergistic as well as antagonistic effects should be taken into account during evaluation of their biological activities. Essential oils can be antimicrobial, cytotoxic or antiseptic. The essential oils can be used as an alternative to antibiotics due to numerous side effects of existing antibiotic drugs (http://www.therapeuticoils.com).

In the present investigation an attempt was made to carry out the study of composition of volatile oils of Morina longifolia collected from Western Himayan region of Uttrakhand.

MATERIALS AND METHODS:

Plant Material:

The fresh aerial parts of M. longifolia were collected from Milam glacier of Kumaun Himalaya at the altitude of 3600m. Plant herbaria were identified in Botanical Survey of India and Forest Research Institute, Dehradun. The voucher specimen (No.CHEM/DST/06/03) has been deposited in the Phytochemistry Research Laboratory, Kumaun University, Nainital.

Oil Isolation:

The fresh plant materials (1.5 kg each) were subjected to steam distillation using a copper electric still, fitted with spiral glass condensers the yields 0.32%. The distillates were saturated with NaCl and extracted with n-hexane and dichloromethane. The organic phase was dried over anhydrous Na₂SO₄ and the solvent was distilled off in a rotary vacuum evaporator at 30^oC.

GC and GC-MS analysis:

The GC analysis was run on Nucon 5765 gas chromatograph (Rtx-5 column, 30 m 0.32 mm i.d., FID), split ratio 1: 48, N₂ flow of 4 kg/cm² and on Thermo Quest Trace GC 2000 interfaced with Finnigan MAT PolarisQ ion trap mass spectrometer fitted with a Rtx-5 (Restek Corp.) fused silica capillary column (30 mm 25 mm; 0.25 μm film coating). The column temperature was programmed from 60^oC-210^o at 3^oC /min. using He as carrier gas at 1.0 mL/min. The injector temperature was 210^oC, injection size 0.1μL prepared in n-hexane, split ratio 1:40. MS were taken at 70 eV with mass scan range of m/z 40-450 amu.

Table 1 Volatile oil composition of the aerial parts of Morina longifolia

S. No	Compounds	RI	%Compo-sition (FID)	Mode of Identifica-tion
1.	n-heptanal	899	2.36	a, b
2.	tricyclene	926	4.34	a, b
3.	α-thujene	931	t	a, b
4.	α-pinene	939	4.84	a, b
5.	camphene	953	0.13	a, b
6.	benzaldehyde	961	t	a, b
7.	sabinene	976	0.11	a, b
8.	β-pinene	980	0.55	a, b
9.	6-methyl-5-hepten-2-one	985	0.24	a, b
10.	2-octanone	988	0.16	a, b
11.	myrcene	991	2.96	a, b
12.	2-octanol	997	0.10	a, b
13.	α-phellandrene	1005	0.67	a, b
14.	α-terpinene	1018	t	a, b
15.	p-cymene	1026	0.34	a, b
16.	β-phellandrene	1031	3.24	a, b
17.	1,8-cineole	1033	0.26	a, b
18.	(Z)-β-ocimene	1040	0.15	a, b
19.	(E)-β-ocimene	1050	0.22	a, b
20.	cis-sabinene hydrate	1068	0.11	a, b
21.	terpinolene	1088	1.01	a, b
22.	trans-sabinene hydrate	1068	0.54	a, b
23.	linalool	1098	1.27	a, b
24.	trans-thujone	1114	0.55	a, b
25.	cis-p-menth-2-en-1-ol	1121	0.21	a, b
26.	chrysanthenone	1123	0.12	a, b
27.	trans-p-menth-2-en-1-ol	1140	t	a, b
28.	borneol	1165	t	a, b
29.	terpinen-4-ol	1177	t	a, b
30.	α-terpineol	1189	0.67	a, b
31.	hexyl butyrate	1191	1.27	a, b
32.	verbenone	1204	0.73	a, b
33.	citronellol	1228	0.89	a, b
34.	bornyl acetate	1285	0.99	a, b
35.	trans-sabinyl acetate	1290	0.13	a, b
36.	(E)-methyl cinnamate	1301	0.77	a, b
37.	δ-elemene	1339	t	a, b
38.	α-cubebene	1351	0.58	a, b
39.	citronellyl acetate	1354	2.73	a, b
40.	β-bourbonene	1384	0.49	a, b
41.	β-cubebene	1390	t	a, b
42.	α-gurjunene	1409	0.12	a, b
43.	β-caryophyllene	1418	1.42	a, b
44.	β-gurjunene	1432	0.16	a, b
45.	α-humulene	1454	0.67	a, b
46.	γ-gurjunene	1473	0.17	a, b
47.	γ-muurolene	1477	t	a, b
48.	germacrene D	1480	10.75	a, b,c
49.	cis-β-guaine	1490	t	a, b
50.	bicyclogermacrene	1495	4.26	a, b
51.	α-muurolene	1499	1.65	a, b
52.	germacrene A	1503	1.93	a, b
53.	cubebol	1514	t	a, b
54.	δ-cadinene	1524	1.23	a, b
55.	germacrene B	1556	0.85	a, b
56.	germacren D-4-ol	1574	1.29	a, b
57.	spathulenol	1576	1.14	a, b
58.	caryophyllene oxide	1581	3.30	a, b
59.	(E)-2-hexenyl benzoate	1583	2.08	a, b
60.	humulene epoxide-II	1606	t	a, b
61.	10-epi-γ-eudesmol	1619	0.95	a, b
62.	γ-eudesmol	1630	0.32	a, b
63.	epi-α-cadinol	1640	1.90	a, b
64.	β-eudesmol	1649	t	a, b
65.	α-cadinol	1653	4.26	a, b
66.	bulnesol	1666	0.85	a, b
67.	(E)-citronellyl tiglate	1667	4.20	a, b
68.	β-bisabolol	1671	1.70	a, b
69.	α-bisabolol	1683	3.02	a, b
70.	(Z,E)-farnesol	1697	0.29	a, b
71.	(E,E)-farnesol	1722	1.57	a, b
72.	curcumenol	1726	0.50	a, b
73.	oplopanone	1733	1.11	a, b
74.	pimaradiene	1941	3.90	a, b
75.	sandaracopimara-8 (14),15-diene	1960	1.07	a, b
Aliphatic compunds			5.97
Monoterpene hydrocarbons			18.56
Oxygenate monoterpenes			14.41
Sesquiterpene hydrocarbon			24.28
Oxygenated sesquiterpenes			22.20
Diterpenoids			4.97
Total identified			90.39

a=Retention Index (RI) on Rtx-5 capillary column, b=MS (GC-MS), c= ¹H NMR ¹³C NMR data,

Compounds >3.0% represented in bold face, t= trace (<0.1%)

The identification was done on the basis of Linear Retention Index (LRI, determined with reference to homologous series of n-alkanes (C₉-C₂₄Polyscience Corp., Niles IL) under identical experimental conditions, co-injection with standards (Sigma), MS Library search (NIST and WILEY), by comparing with the MS literature data(Adams, 1995).

RESULTS AND DISCUSSION:

The GC and GC-MS analysis of leaf oil of Morina longifolia, identified constituents of the oil are listed in Table 1 in order of their elution in Rtx-5column. The major constituents in leaf oil were germacrene D (10.75 %) α-pinene (4.84 %), bicyclogermacrene (4.26 %), α-cadinol (4.26 %), (E)-citronellyl tiglate (4.20 %) β-phellandrene (3.24 %). Morina species showed that, a new aromatic glycoside characterized as 2,6-dihydroxy-5-methoxy-(3-C-glucopyranosyl) benzoic acid was isolated along with four known compounds from the aerial parts of Morina longifolia (Bodakhe et al., 2010). Previous study of essential oil of M. longifolia showed as a totally different composition as β-myrcene as major constituents has been reported from different regions (R.S. Chauhan et al., 2012; D. K. Semwal et al., 2010). We can see that the results of our work is totally different with previous one, this also showed that the oil composition vary with different regions.

ACKNOWLEDGMENT:

The authors are grateful to the Department of Science and Technology (DST) New Delhi for GC-MS grant. We are also thankful to BSI, Dehradun for the identification of the plant.

REFERENCES:

1. Adams, R.P, 1995.Identification of Essential Oil Components by Gas Chromatography/ Mass Spectroscopy. Allured Publishing Corporation. Carol Stream, IL., USA.

2. Bodakhe SH, Ram A, Pandey DP. A new aromatic glycoside from Morina longifolia Wall. Asian Journal of Chemistry. 2010; 22: 2789-93.

3. Chopra, R.N., Nayer, S.L.,Chopra, I.C., 1956. Glossary of Indian Medicinal Plants, pp.105, Council of

4. Industrial and Scientific Research, New Delhi.

5. Gaur, R.D., 1999. Flora of District Garhwal North West Himalaya (With ethno botanical notes). 1st ed. Srinagar Garhwal, India, Trans Media, pp.550.

6. Handoo S. 2006 A surveys of plants used for wound healing in animals, Veterin J 1:2.

7. Polunin, O., Stainton, A., 1984. Flowers of the Himalaya,Oxford University Press, New Delhi.

8. Tsarong, T. J., Tibetian., 1994. Medicinal Plants, pp.94, Tibetian Medical Publications, New Delhi.

9. Tang, R., Xie, H., Liu, X., Wang, D. and Yang, C., A novel acylated flavonol glycoside from Morina nepalensis var. alba. Fitoterapia, 2002, 73, 1.

10. Su, Bao-Ning., Takaishi, Yoshihisa 1999 Morinins L—P, Five New Phenylpropanol Derivatives from Morina chinensis Chem. Pharm. Bull. 47(11) 1569—1572) 1569

11. Su Bao-Ning, Takaishi Yoshihisa, Duan Hong-Quan, Chen Bei, Phenylpropanol Derivatives from Morina chinensis J. Nat. Prod. 1999, 62, 1363-1366

12. Su Bao-Ning, Takaishi Yoshihisa Kusumi Takenori, 1999 Twelve Novel Sesquineolignans and Neolignans Morinols A-L, With a New Carbon Skeleton isolated from Marina chinensis Tetrahedron 55 (1999) 14571-14586

13. S.K. Bhattacharjee, Handbook of Aromatic Plants. Pointer Publisher, Jaipur (2000).

14. www.therapeuticoils.com

Received on 26.12.2012 Accepted on 02.02.2013

Asian J. Res. Pharm. Sci. 3(1): Jan.-Mar. 2013; Page 12-14