Yu. I. Korniyevsky, V. G. Korniyevskaya, S. V. Panchenko

Zaporizhya State Medical University

CHROMATO-MASS-SPECTROMETRY OF AN ESSENTIAL OIL OF VALERIANA OFFICINALIS L.s.l, WEST OF UKRAINE

Key words:Valeriana officinalis, gas chromatography, chromato-mass spectrometry, essential oil.

By the method of comparative gas chromatography in examples of V. stolonifera Czern essential oil, collected in Zaporizhya region, had found 91 components, in V. grossheimii Worosch, collected in Crimea,- respectively 66. By means of chromato-mass spectroscopy in essential oils identified the row of components of: V. stolonifera Crern-30 chemical compounds, V. grosshemii Worosch -25 chemical compounds. The similarity of oils counts 10 components, the difference of two examples of essential oils composition the next: V. stolonifera Crern -81 components ,V. grosshemii Worosch -56 accordingly.

The main components of the essential oils of V. stolonifera Crern -2,2-dimethyl-3-methylene-(1R)-bicyclo[2.2.1] heptane (12,35%); 1,7,7-trimethyl- (1S-endo)bicyclo[2.2.1] heptane-2-ol (7,66%); 1Sα-pinene (4,3%); d-limonene (2,4%); trans-α-bergamoten (2,23%); V. grosshemii- 1,7,7-trimethylbicyclo[2.2.1]hept-2-ilacidum acetic (9,43%); 1,7,7-trimethyl-(1S-endo) bicyclo[2.2.1]heptane-2-ol (3,8%); dihidroyasmon (2,17%).

Valeriana officinalis is a sectional species,to composition of which in Ukraine enter 13 species, including the most common on the southof Ukraine: V. stolonifera Crern and endemic of the Crimea - V. grosshemii Worosch.

Valeriana officinalis as a medical plant has an old history of it's using by humanity.

Beginning from XIX century, the biological activity of Valeriana associated with an essential oil [4, 9].The essential oil's amount in Valeriana depends on the species of a plant, it's age ecology, harvesting time, drying conditions and conservation of a plant [2, 3]. Moreover had published the raw of works with information about the existence of wild growing Valeriana’s species with the large number of different chemotypes (the plants, which have the same form, but significantly differs in essential oils composition) [10]. Thereby, is continuing the research of essential oil’s comparison of the Valeriana’s radix by scientists of different countries of the world [5 – 10].

Moreover,in accordance with the latest facts the neuro-mediatory activity of plants may be caused by valeriana’s acids [4, 7 - 9].

Tranquilizing valerianas effect is connected with valepotriates,which are promoting to the elimination of worry and fear, help if you have an insomnia.This substances are the classic hypnotic phytotranquilisators,which are show(as like the most common for the frequency of using preparations- benzodiazepine derivatives) the tranquilizing properties, what is expressed mainly in anxiolytic, antiphobic,anticonvulsant, antiagressive, antidepressant, antistres effects. Valeriana accelerates the dreaming processes, makes dream deeper, prolongs on 30-50% the effects of other dormitive preparations, potentiates the influence of sedative, anticonvulsant and other preparations.

Experiments on animals show the Valeriana’s ability to renders a sedative effect on central nervous system, and also, it's property to prevent spasms. Conducted the greate ammount of clinical researches, expecially in heriartry, with the participation of patients,which suffered from insomnia and nervous disorders. As a rule, valeriana’s drugs reduce the time of falling asleep and improve dream's quality [6].Valeriana is recommended for the treatment of neuro-vegetative disorders and light dream's violations as at adults,as at child’s.Valeriana often uses together with other sedative preparations, for example with pasiflora and hawthorn.

In accordance with nowadays facts a chemical composition of an essential oil of the Valeriana officinalis is very complex, have already identified 84 substances [10]. During the research of different examples of view of Valeriana's collectional sort was established three hemotypes of an essential oil, which differs from each other by it's quantitative composition of components,that had confirmed by different researches.

Aim of the work: by means of gas-fluid chromatography with mass-spectrometry detector to study a composition of essential oil of Valeriana stolonifera Crern and Valeriana grosshemii Worosch.

Experimental part. Essential oils from the raw examples received by method of distillation with water vapor in accordance with SPU 1,2 [1]. In a flack, with volume 2000ml added 500ml of purified water and 40g of fresh, shredded in a powder raw. Distillation was performed with speed from 3ml/min to 4ml/min during 4 hours.Received essential oils dissolved in 5 ml of hexane, conducted reaction with dehydrate sodium sulfate and researched by capillary gas- chromatography method with mass-spectrometry detection. The essential oil’s content in our examples was 1,20% for Valeriana stolonifera Crern and 1,09% for Valeriana grosshemii Worosch.

Received examples of essential oils were chromatographied on gas chromatograph with seria 6890 N of "Agilent Technologies" production (injector 7883 B; mas-selective defector 5975).

Chromatography terms: sample volume -1mcl ,a column -DB-WaX30 m x 0,25 mm 0,25 mkm with Makrogol 20000 sorbent with layer thickness 0,25 mkm; the column temperature- 40°C 9 min , temperature increase 4°C/min up to 250°C, firmness at 250°C 9 min. The evaporators temperature – 300°C , the stream division(helium) = 1:20; V(He)=1,9 ml/min(52sm/c)

Detection: electronic-shock ionization (70 eB); temperature of the ionization camera 230°C, quadrupole temperature 150°C . Identification of component’s examples performed by means of NISTO5a spectra’s library.

The results of the components identification of two examples of essential oils are presented in table 1. Contents of the individual components in essential oils examples estimated by normalization method(the peak area represented in percents up to sum of all peak’s areas on the chromatogram of an example, except the peak of a solvent).

Table 1

The results of chromato-mass spectrometry identification of essential oils of valeriana

№	The retention time, min	Component	Peak area, %
			Valerianastolonifera	Valeriana grossheimii
1	2,38	3-Methyl-butanal	0,00089
2	3,21	1,7,7-Trimethyl-bicyclo[2.2.1]hept-2-en	0,075
3	3,73	1,7,7-Trimethyl-tricyclo[2.2.1.0(2,6)]heptan	0,11
4	4,05-4,17	1 S α-Pinene	4,3	0,152
5	4,97	7,7-Dimethyl-2-methylene-bicyclo[2.2.1] heptan		0,3
6	5,19	Camphene	0,03	0,372
7	5,53	2,2-Dimethyl-3-methylene-(1R)-bicyclo[2.2.1]heptan	12,35
8	6,60	6,6-Dimethyl(1S)bicyclo[3.1.1]heptan		0,1
9	7,01	2,2-Dimethyl-3-methylene-(1S)-bicyclo[2.2.1]heptan	1,74
10	10.80; 23,31-23,54	1-Methyl-4-(1-methylethyl)-cyclohexa-1,3-diene	0,12 1,32	1,69
11	11,68-12,00	d-Limonene	2,4	0,118
12	12,05	β-Phelandren		0,042
13	14,12-14,28	1-Methyl-4-(1-methylethyl)-cyclohexa-1,4-diene	0,42	0,018
14	15,17-15,29	1-Methyl-2-(1-methylethyl)-benzen	0,7	0,032
15	15,78; 16,87	3-Methylbutyl esther de 3-methyl butanoic acid	0,18	0,008
16	15,90	1-Methyl-4- (1-methilethiliden) -cyclohexen	0,17
17	19,30	Hexanol-1		0,006
18	20,07	1,3,3-Trimethyl-bicyclo[2.2.1] heptan-2-on		0,06
19	27,03	1,7,7-Trimethyl-bicyclo[2.2.1] hept-2-il acetic acid		9,43
20	27,07; 30,37	1,7,7-Trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol	7,66	3,8
21	27,66	4-Methyl-1- (1-methylethyl)-cyclohex-3-en-1-ol	0,49
22	28,95	4,6,6-Trimethyl-(1S)-bicyclo[3.1.1]hept-3-en-2-on	1,1
23	29,34	3-Methylbutanoic acid	1,28	0,65
24	29,87	(-)-Mirtenil acetate		1,52
25	30,36	2-Methyl-1-methylene-3-(1-metiletinil)-cyclopentan	0,39
26	30,45	Borneol	0,55
27	30,49-30,57	1-Ethyl-3-vinyl-adamantane	0,32	1,04
28	31,25	Trans-α-bergamoten	2,23
29	32,79-32,91	Bicyclo[3.1.1] hept-2-en-2- methanol-6,6-dimethyl	1,58	0,71
30	33,46	1-Methoxy-4-(1-propenyl)-benzen		0,27
31	33,52	Izoftaldiamidoksim	0,21
32	35,23	Esther phenylmethylpentenoic acid		0,44
33	35,65	Cis-2-methyl-5-(1-methylethyl) cyclohex-2-en-1-ol acetat	0,1
34	36,46-36,53	3-Buten-2-on,4-(2,6,6- trimethyl-1-cyclohexen-1-il)-,(E)	1,3	1,92
35	37,25	Dihydroyasmon		2,17
36	37,96	Caryophyllene oxide		0,4
37	39,00	2-Methyl bicyclo[2.2.2]octane	0,11
38	42,44	2-Methyl-5-(1-methylethyl)-phenol	0,55
39	45,96	2,2a,3a,4,6a,6b-Hexahydro-3a-methyl-1,2,4-meteno-1H-cyclobuta[b]cyclopenta[d] furan	0,08
40	49,20	1-Methyl-4-(2-methyloxyranil)-7-oxabicyclo[4.1.0]heptane	0,48
41	49,46	(+,-)-1,3,3-Trimethylcycloheks-1-en-4-karboksalydegid		0,26
42	54,06	Acidum benzene aceticestherphenylmethyl		0,1
43	62,03	Tetradesin		0,3

Some of the components included in the two kinds of valerianа: α-Pinene (4,3%; 0,152%); сamphene (0,03%; 0,372%); 1-methyl-4-(1-methylethyl)-cyclohexa-1,3-diene (0,12%; 1,32%; 1,69%); d-Limonene (2,4%; 0,118%); 1-methyl-4-(1-methylethyl)-cyclohexa-1,4-diene (0,42%; 0,018%); 1-methyl-2-(1-methylethyl)-benzen (0,7%; 0,032%); 3-methylbutyl esther de 3-methyl butanoic acid (0,18%; 0,008%); 1,7,7-trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol (7,66%; 3,8%); 3-methylbutanoic acid (1,28%; 0,65%); 1-ethyl-3-vinyl-adamantane (0,32%; 1,04%); bicyclo[3.1.1] hept-2-en-2- methanol-6,6-dimethyl (1,58%; 0,71%); 3-buten-2-on,4-(2,6,6- trimethyl-1-cyclohexen-1-il)-,(E) (1,92%; 1,3%).

In the essential oil of Valeriana stolonifera it found 91 components, Valeriana grossheimii– 66 components.

Using chromato-mass-spectrometry in the essential oil of Valeriana grossheimii had found:α-Pinene; 7,7-Dimethyl-2-methylene-bicyclo[2.2.1] heptan; Camphene; 6,6-Dimethyl(1S)bicyclo[3.1.1]heptan; 1-Methyl-4-(1-methylethyl)-cyclohexa-1,3-diene; d-Limonene; β-Phelandren; 1-Methyl-4-(1-methylethyl)-cyclohexa-1,4-diene; 1-Methyl-2-(1-methylethyl)-benzen; 3-Methylbutyl esther de 3-methyl butanoic acid; Hexanol-1; 1,3,3-Trimethyl-bicyclo[2.2.1] heptan-2-on; 1,7,7-Trimethyl-bicyclo[2.2.1] hept-2-il acetic acid; 1,7,7-Trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol; 3-Methylbutanoic acid; (-)-Mirtenil acetate; 1-Ethyl-3-vinyl-adamantane; Bicyclo[3.1.1] hept-2-en-2- methanol-6,6-dimethyl; 1-Methoxy-4-(1-propenyl)-benzen; Esther phenylmethylpentenoic acid; 3-Buten-2-on,4-(2,6,6- trimethyl-1-cyclohexen-1-il)-,(E); Dihydroyasmon; Caryophyllene oxide; (+,-)-1,3,3-Trimethylcycloheks-1-en-4-karboksalydegid; Acidum benzene aceticestherphenylmethyl; Tetradesin.

Using chromato-mass-spectrometry in the essential oil of Valeriana stolonifera had found:3-Methyl-butanal; 1,7,7-Trimethyl-bicyclo[2.2.1]hept-2-en; 1,7,7-Trimethyl-tricyclo[2.2.1.0(2,6)]heptan; α-Pinene; Camphene; 2,2-Dimethyl-3-methylene-(1R)-bicyclo[2.2.1]heptan; 2,2-Dimethyl-3-methylene-(1S)-bicyclo[2.2.1]heptan; 1-Methyl-4-(1-methylethyl)-cyclohexa-1,3-diene; d-Limonene; 1-Methyl-4-(1-methylethyl)-cyclohexa-1,4-diene; 1-Methyl-2-(1-methylethyl)-benzen; 3-Methylbutyl esther de 3-methyl butanoic acid; 1-Methyl-4- (1-methilethiliden) –cyclohexen; 1,7,7-Trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol; 4-Methyl-1- (1-methylethyl)-cyclohex-3-en-1-ol; 4,6,6-Trimethyl-(1S)-bicyclo[3.1.1]hept-3-en-2-on; 3-Methylbutanoic acid; 2-Methyl-1-methylene-3-(1-metiletinil)-cyclopentan; Borneol; 1-Ethyl-3-vinyl-adamantane; Trans-α-bergamoten; Bicyclo[3.1.1] hept-2-en-2- methanol-6,6-dimethyl; Izoftaldiamidoksim; Cis-2-methyl-5-(1-methylethyl) cyclohex-2-en-1-ol acetat; 3-Buten-2-on,4-(2,6,6- trimethyl-1-cyclohexen-1-il)-,(E); 2-Methyl bicyclo[2.2.2]octane; 2-Methyl-5-(1-methylethyl)-phenol; 2,2a,3a,4,6a,6b-Hexahydro-3a-methyl-1,2,4-meteno-1H-cyclobuta[b]cyclopenta[d] furan; 1-Methyl-4-(2-methyloxyranil)-7-oxabicyclo[4.1.0]heptane.

The main components the essential oils of V. stolonifera Czern: 2,2-Dimethyl-3-methylene-(1R)-bicyclo[2.2.1]heptan (12,35%); 1,7,7-Trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol (7,66%); 1 S α-Pinene (4,3%); d-Limonene (2,4%); Trans-α-bergamoten (2,23%);

The main components the essential oils of V. grossheimii: 1,7,7-Trimethyl-bicyclo[2.2.1] hept-2-il acetic acid (9,43%); 1,7,7-Trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol (3,8%); Dihydroyasmon (2,17%). This distinctive components of valerianastolonifera and valerianagrossheimii.

Conclusions

1. By comparison gas-liquid chromatography method in researched examples of an essential oil of V. stolonifera Czern revealed 91 specific components, and in V. drossgeimii Worosch accordingly 66.

2. By means of chromate-mas-spectroscopy was identified in essential oil of V. stolonifera Czern – 30 chemical compounds, V. drossgeimii Worosch – 25 chemical compounds.

3. The similarity of essential oils counts 10 components, the composition difference of two examples of essential oils consists V. stolonifera Czern – 81 components, and V. drossgeimii Worosch accordingly 56.

4. The main components the essential oils of V. stolonifera Czern: 2,2-Dimethyl-3-methylene-(1R)-bicyclo[2.2.1]heptan (12,35%); 1,7,7-Trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol (7,66%); 1 S α-Pinene (4,3%); d-Limonene (2,4%); Trans-α-bergamoten (2,23%);V.grossheimii: 1,7,7-Trimethyl-bicyclo[2.2.1] hept-2-il acetic acid (9,43%); 1,7,7-Trimethyl-(1S-endo)bicyclo[2.2.1] heptan-2-ol (3,8%); Dihydroyasmon (2,17%).

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