Biologically active substances of aqueous-ethanolic extract of Tagetes signata Bartl. № 13152-8 „Vetvisty‟ specimen of nikita Botanical Gardens collection Текст научной статьи по специальности «Фундаментальная медицина»

PLANT BIOCHEMISTR Y

UDK 582.998.1:577.19 (477.75)

BIOLOGICALLY ACTIVE SUBSTANCES OF AQUEOUS-ETHANOLIC EXTRACT OF TAGETESSIGNATA BARTL. № 13152-8 'VETVISTY' SPECIMEN OF NIKITA BOTANICAL GARDENS COLLECTION

Gury Viktorovich Kornilyev, Anfisa Yevgenyevna Paly, Ivan Nikolayevich Paly, Valery Dmitriyevich Rabotyagov, Sergey Aleksandrovich Feskov

Nikita Botanical Gardens - National Scientific Centre 298648, the Republic of Crimea, the city of Yalta, urb.vil.Nikita [email protected]

Introduction

Tagetes L. genus includes more than 30 cultivars growing in vivo, mainly in Central America. Tagetes patula L. Tagetes erecta L., Tagetes signata Bartl., syn. Tagetes tenifolia Cav. are considered the most popular within crop [5, 18, 27].

Different Tagetes cultivars are used to treat liver or kidneys disorder, for pain control - as a diuretic and pathogenic remedy. Tagetes L. preparations permit to control nervous tension and improve spirit. The plant is popular in dermatology as well, because it cures blackheads and possesses tonic and regenerative properties, so important for skin [1, 10, 13, 16, 17, 19, 21, 26, 30].

Essential oils (EO) of different Tagetes L. cultivars are known as matters with antibacterial, antimicrobial, larvicidal, repellent and fungicidal effect [7, 10, 11, 22-24, 26, 28, 29].

Tagetes flowers are possible to use as a spice "Imeretian Saffron" [30]. Besides some species of Tagetes L. are source of carotenoids (for example, in poultry farming - for intensifying egg color) [2, 4, 9], pigments and lutein [3, 20].

Allowing for variety of useful Tagetes L. properties, introduction of new cultivars is of great importance now, particularly on South Coast of the Crimea, as well as breeding new sorts with valuable characteriristics.

Nikita Botanical Gardens - National Scientific Centre bred promising (by ornamentality, disease- and pest-resistance, crop capacity, essential oil content) specimenTagetes signata L. № 13152-8 "Vetvisty". It was found out that essential oil composition of this specimen included trans-tagetenon, dihydro-tageton, trans-tageton, cis-tagetenon, limonene, sabinene, trans-ocymen, eugenol and a number of minor components [15].

In this way EO extracting out of raw material is accompanied with oxidation process if to apply method of steam distillation; getting native substances demands their transition into water-ethanol extracts.

It's a well-known fact, that flower extracts of T.signata is a result not only of volatile substances transition, but phenol substances, which present antioxidant properties: hydroxycoric acids (caffeic, rosemary, chlorogenic, p-Coumaric acid), flavonoids and their glycosides (apigenin, hyperoside, isoquercetin, quercetin, rutin) [12, 15]. At the same time there is dependence of chemical composition in raw material from cultivar and sort belonging, soil and climatic conditions of habit.

Allowing for data mentioned above, investigation of chemical composition of T.signata specimen water-ethanol extract, № 13152-8 "Vetvisty", cultivated on South Coast of the Crimea, is a vital question.

The study purpose is to research qualitative and quantitative composition of biologically active substances (volatile terpenes, phenol compounds, vitamins) of T. signata № 13152-8 "Vetvisty" under conditions of South Coast of the Crimea, to get an opportunity for use it in medioprophylactic production.

Objects and methods of the research

Study research is water and ethanol extract out of specimen T.signata №13152-8 "Vetvisty" flowers, yielded during mass blooming.

Specimen was bred in NBG-NSC applying individual selection from seed population. Plant of 50-60 sm high. Stem is straight, branchy from the bottom, well-leafed. Leaves are sectile. Inflorescence is a basket of 19-24 mm across diameter, 5 semiflosculous flowers, corolla is golden-yellow. Blooming period is the whole summer, mass blooming happens from July the 10-20th. Specimen is disease- and pest-resistant. Crop capacity made 84,7 center/ha, essential oil content - 0,46% of the raw material, EO yield made 40,8 kg per hectare.

Concentration of biologically active substances was determined in water-ethanol extract, made of air-dry plant mass, yielded during mass blooming. Extraction was carried out applying 50-% ethanol having ratio of raw material and leach - 1:10 what was prepared for 10 days at room temperature.

Component composition of volatile substances was determined applying chromatograph Agilent Technology 6890N with mass-spectrum detector 5973N. Chromatographic column - capillary HP-1 (30 m long); inside diameter - 0,25 mm. Thermostat temperature was programmed in a range 50-250 0C with velocity of 40C/min. Injector temperature - 2500C. Carrier gas is helium, stream velocity - 1,0 sm3/min. Transfer from gas chromatograph to mass spectrometric detector was heated till 2300C. Source temperature was kept up at 2000C. Electronic ionization was carried out at 70 eV with mass range m/z from 29 till 450. Identification of essential oil components was guided by comparison results obtained in terms of chemical substances mass spectra with publishing data of mass spectra NIST05-WILEY2007 (about 500000 of mass spectra).

Component composition of phenol substances was determined applying chromatograph "Agilent Technologies" (model 1100), completed with running vacuum degasifier G1379A, 4-channeled pump of low pressure gradient G 13111A, automotive injector G 1313A, thermostat of heaters G13116A, diode-matrix detector G1316A, fluorescent detector G1315B. Chromatograph heater 2,1mm x 150 mm, filled with octadecilsilil sorbent "ZORBAX" SB-C18 and 3,5 mkm graining was applied during analysis. Gradient regime of chromatography that forsees changes of component A (0,1% orthophosphoric acid, 0,3%-tetrahydrofuran, 0,018%- triethylamine) and B (methanol) components in eluting mixture. Feed rate of mobile phase made 0,25 sm3/min; working pressure of eluent - 240-300 kPa; sample volume - 2 mkl; scanning time 2 sec, scale of measurements - 1,0. Identification of phenol substances was carried out according to time of standard keeping and spectrum characteristics (wave length - 313 (for phenol acids and their derivatives), 350 (for glycosides of flavonoids), 371 nm (for flavonoids)).

Concentration of carotinoids was determined applying photometric method [14], ascorbic acid - by titration of potassium iodate [6].

Results and discussion

It was found out that concentration of volatile substances in specimen extract (T.signata № 3152-8 "Vetvisty") made 32,96 mg/100 g of raw material. 7 components were identified (table 1, fig.1).

Table 1

Volatile substances of water and ethanol extract Tagetes signata Bartl. № 13152-8 "Vetvisty"

№ Output time, min Component Concentration, %

1 6.38 cyclohexanone 8,49

2 7.25 3,5 -dimethylcyclohexe-2-en- 1-on 10,2

3 8.27 propylvaleriate 2,28

4 8.36 hexe-1,2,6-triol 2,76

5 14.12 tagetenone 17,3

6 15.11 4-vinylphenyl acetate 8,23

7 16.63 P-vinylguaiacol 50.8

The principal volatile components of this extract are P-vinylguaiacol (50,8%) and typical for T.signata № 13152-8 "Vetvisty" tagetenone (17,3%). Dominant group of compounds in the extract is aroma (59%; P-vinylguaiacol and 4-vinylphenyl acetate). The biggest number of components (3,5-dimethylcyclohexe-2-en-1-on, cyclohexanone, hexe-1,2,6-triol, propylvaleriate) belongs to aliphatic compounds, what is 23,7% in amount. Typical for T.signata component - tagetenone is a monoterpenic compound.

Fig. 1 Chromatogram of volatile substances in water-ethanol extract of Tagetes signata Bartl. № 13152-8

"Vetvisty"

It was found out that content of phenolic substances in extract of T.signata № 131528 "Vetvisty" made 1821 mg/100 g of plant material (table 2, fig.2). 11 components were revealed, 6 compounds - identified.

Table 2

Phenolic substances of water and ethanol extract Tagetes signata Bartl.№ 13152-8 "Vetvisty"

№ Output, min Component Concentration mg/100 g

1 15.35 Caffeic acid derivatives 142

2 17.92 Ferulic acid 152

3 18.73 Unidentified flavonoid 132

4 18.96 Unidentified flavonoid 79

5 19.34 Quercetin -3-O-galactoside 301

6 19.92 Rutin 331

7 20.28 Unidentified flavonoid 65

8 20.76 Unidentified flavonoid 332

9 21.40 Quercetin -3-O-glucoside 217

10 23.70 Unidentified flavonoid 38

11 24.39 Lutheoline 32

Among identified substances in study extract the following dominate: glicosides of quercetin - rutin (331), quercetine-3-O-galactoside (301) and quercetine-3-O-glucoside (217 mg/100 g). Hydroxycoric acids are presented by ferulic acid and caffeic acid derivatives, which make 294 mg/100 g in total.

DAD1 C, Sig=350,32 Ref=off (HAR014-3\PHEN0034.D) mAU

.^yv/V^wvJ

Fig.2 Chromatogram of phenolic substances in water-ethanol extract of Tagetes signata Bartl. № 13152-8

"Vetvisty"

Vitamin concentration was determined in the study extract as well. Concentration of ascorbic acid made 14,54 mg/100 g, carotinoids - 5,19 mg/100 g.

2

1 50

5

8

6

1

56

0

5

20

25

m n

In this sway water-ethanol extract of T.signata specimen №13152-8 "Vetvisty" contains not so much of volatile terpenes, but could be considered as a source of P-vinylguaiacol, glicosides of quercetine, ascorbic acid and carotinoids.

Conclusions

In terms of the research qualitative and quantitative composition of biologically active substances (volatile terpenes, phenolic substances and vitamins) in water-ethanol extract of Tagetes signata Bartl. specimen, № 13152-8 "Vetvisty", cultivated on South Coast of the Crimea.

It was found out that among volatile compounds of the specimen, P-vinylguaiacol and tagetenone dominate.

At the same time among phenolic substances glicosides of quercetine (rutin, quercetine-3-O-halactoside, quercetine-3-O-glucoside) and flavonoids prevail. Hydroxycoric acids are presented by ferulic acid and derivatives of coffeic acid.

In extract of T.signata № 13152-8 "Vetvisty" we determined vitamin concentration -ascorbic acid and carotinoids.

In general specimen of T.signata Bartl. № 13152-8 "Vetvisty" could be considered as a source of biologically active substances, especially of P-vinylguaiacol, glicosides of quercetine, ascorbic acid and carotinoids.

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The article was receaived at editors 1.02.2016

Kornilyev G.V., Paliy А.Ye., Rabotyagov V.D., Feskov S.A. Biologically active substances of aqueous-ethanolic extract of Tagetes signata Bartl. № 13152-8 'Vetvisty' specimen of Nikita Botanical Gardens collection // Bull. of the State Nikit. Botan. Gard. - 2016. - № 118. - P. 39-45.

The qualitative and quantitative composition of Tagetes signata Bartl. № 13152-8 'Vetvisty' specimen was investigated in terms of the research. It was established that among volatile substances there were aromatic (59%; p- vinylguaiacol prevails), aliphatic (23,7 %) and monoterpenoide (tagetenon content is 17,3%) compounds. Phenolic compounds were presented by flavonoids and their glycosides (rutin, quercetin-3-O-galactoside, quercetin-3-O-glycoside), and hydroxycinnamic acids (caffeic, ferulic). The ascorbic acid (14,54) and carotenoids (5,19 mg/100 g) were identified as well. On the whole, the specimen T. signata № 13152-8 "Vetvisty" can be considered as a sources of biologically active substances such as p- vinylguaiacol, flavonoid glycosides, ascorbic acid and carotenoids.

Key words: Tagetes signata Bartl.; specimens; aqueous-ethanolic extract; volatile substances; phenolic substances; vitamins.

PHYTOMONITORING

UDK 635.055:504.753:712.253

LOWER LAYER ORNAMENTAL PLANTS OF ARBORETUM IN NIKITA BOTANICAL GARDENS AND THEIR GROWTH AND STATUS DEPENDING ON

ILLUMINATION CONDITIONS

Nikolay Ivanovich Klymenko, Ivan Nikolayevich Paly

Nikita Botanical Gardens - National Scientific Centre 298648, the Republic of Crimea, the city of Yalta, urb.vil.Nikita [email protected]

Introduction

Investigation of shade tolerance of ornamental plants, peculiarities of their light conditions plays a big role for matching of cultivars and crop combinations projecting ladscape compositions. Shade-tolerant plants have wide ecological amplitude; concerning light, good illumination favors their better growth, but at the same time they adapt to weak light easily. As a rule they are divided into more and less shadow-tolerant. Those trees and bushes that grow on open areas and don't stand continuous shade can be classified as less shadow tolerant (light-requiring) arboreal crops. Photosynthesis gets the highest point if good sun illumination. This group includes: birch, willow, larch, aspen, walnut, locust, pine, ash. More shadow tolerant arboreal plants are presented by trees and bushes standing some shadow, but intensively growing under conditions of good illumination: box tree (Buxus), arrowwood (Viburnum), Euonymus Japonicus, cherry laurel, Japan laurel (Aucuba japonica),