INTRODUCTION:

Due to the increase in allergic reactions to synthetic drugs, there has been a demand to expand the range of medications based on medicinal plant-based raw materials. Tanacetum vulgare is a medicinal plant, that contains a large amount of biologically active substances and is widely used in modern and traditional medicine.

Medicinal plant-based raw materials tansy flowers (Flores Tanaceti in Latin) are included in the pharmacopoeias of Belgium, Finland, Portugal (decoction, infusion) as an anthelmintic agent^1,2. Tansy flowers collected at the beginning of flowering are normally used in medicine. Dried individual flower baskets or shields with a peduncle no longer than 4cm (from the upper baskets) can also be used¹. Medications from tansy flowers stimulate appetite, improve digestion, can be used for liver and intestinal diseases, for bronchial asthma, rheumatism, as an anthelmintic for ascariasis and pinworms (infusion), as a medication to increase the acidity of gastric juice. It also relieves the general symptoms for migraines⁹ and can be used as an antibacterial agent. Antiviral and antioxidant activity of tansy flowers is widely studied^4,11,15. The study of Tanacetum vulgare as a callus culture for obtaining biologically active substances in vitro is promising⁵. It has been claimed that the essential oil of Tanacetum vulgarehas insecticidal activity, especially against Leptinotarsa decemlineata, which is very important for the development of potato growing⁸.

Fresh raw tansy has shown to be effective against Acrobasis advenella¹⁰. The toxicity of lyophilized aqueous extraction from tansy leaves has been studied in detail on rats. It is proved that these drugs do not have high toxicity⁷. Any toxic effects of extracts from tansy are associated with the content of thujone in the raw material¹⁴. In the experiment, the antiparasitic properties of the extract from tansy flowers were studied, which turned out to be especially effective in the mass destruction of fish fry by parasitic infusoria Ichthyophthirius multifiliis¹³. An aqueous infusion is prepared from dried flowers. It is taken 1 tablespoon 3 times a day. Tansy preparations are contraindicated during pregnancy.

Flowers of Tanacetum vulgare are part of choleretic collections. Preparations of Tanacetum Vulgare containing the sum of flavonoids and phenol-carboxylic acids are used as choleretic agents⁹. A large number of pectins were found in tansy¹². All the vegetative parts of the plant contain relatively large amounts of terpenoids. The composition of terpenoids can vary greatly depending on climatic conditions, soil fertility and fertilizers applied⁶. The composition and amount of terpenoids can vary significantly in tansy depending on the genotype of the plant⁶.

In the pharmacopoeias of various countries there are articles on medicinal plant-based raw “Flores Tanaceti vulgaris”, however during the harvesting of the medicinal raw material, all the remaining aerial vegetative part is not used, which is an irrational use of a natural resource.

The aim of the study was to study the chemical composition and pharmacological activity of extracts (essential oil, tincture and petroleum extract) obtained from medicinal plant raw materials of tansy grass.

MATERIALS AND METHODS:

The objects of the study were tansy essential oil obtained from fresh raw materials, petroleum extraction of tansy and tincture obtained using the maceration method. Tanacetum vulgare was harvested in the botanical garden of the SechenovFirst Moscow State Medical University during the flowering phase. Petroleum extraction was obtained by maceration with a ratio of raw materials and extractant of 1:10. The tincture was obtained by maceration with a ratio of raw materials and extractant of 1:10. To obtain the essential oil, two different methods of hydrodistillation were used – the Ginsberg method and the Clevenger method. In the Clevenger method, unlike the Ginsberg method, the receiver is placed outside the heating zone, as a result of which the oil is less exposed to temperature.

The qualitative composition of essential oil, tincture and petroleum extraction of tansy was determined by chromatography-mass spectrometry.

Chromatography-mass spectrometric study was carried out with an Agilent Technologies device. Separation of essential oil components was carried out by gas chromatography using a 7890 gas chromatograph (HP column- 5.50m x 320mkm x 1.05 microns). The components were detected using a 5975°C mass-selective detector with a quadrupole mass analyzer. Temperature chromatography program: at 40°C – isotherm for 2 minutes; then programmable heating up to 250°C at a speed of 5°C/min and at 250°C - isotherm for 15 min; then programmable heating up to 320°C at a speed of 25°C/min and at 320°C - isotherm 5 min. Sample input of 1µl. An injector with a 1:50 flow division. The injector temperature was 250°C. Interface temperature was 280°C. The carrier gas was helium; the flow rate is 1ml/min. The chromatogram of the samples was based on the total ion current. Software - ChemStation E 02.00. Conditions of mass spectrometric analysis were: energy of ionizing electrons -70eV; registration of mass spectra in positive ions in the range (m/z) from 20 to 450 at a rate of 2.5scans/sec. The identification of the component composition (qualitative analysis) was carried out using the NIST-05 library of complete mass spectra and the corresponding values of Kovacs chromatographic indices. The relative content of the components of the mixture (quantitative analysis) is calculated from the ratio of the areas of chromatographic peaks according to the total ion current (by simple normalization).

For a model assessment of antioxidant activity, an amperometric method for measuring the total content of antioxidants was used, implemented in the Tsvetyauza-01-AA device. The antioxidant activity of raw materials is associated with the content of flavonoids and essential oil. The point of the method is to register the electric current that occurs during the oxidation of the substance under study (or a mixture of substances) on the surface of the working electrode at a certain potential (from 0 to 1.3 V) At this potential, the oxidation of groups occurs in phenolic and other types of antioxidants. Electrochemical oxidation, proceeding according to the scheme: R-OH → R-O • + e- + H+ (R is the residue of a natural antioxidant: flavonoids, tannins, terpenoids, etc.), can be used as a model process when measuring the ability of substance molecules to intercept free radicals in the biosystem. During the study of antioxidant activity, during extraction, the samples were heated to a temperature of 40°C.

RESULTS:

Monoterpenes, sesquiterpenes, triterpene saponins, fatty acid esters, steroid compounds and vitamins – provitamin A and vitamin D derivative-have been identified in the essential oil obtained by steam distillation. By chromatography-mass spectrometry, 87 compounds were identified in essential oil, from which marker compounds such as camphor, borneol, ascaridiol, cubenol, limonene, carvacrol, pinene, cineol, terpineol and chrysanthenone can be isolated. The relative content of the most important identified compounds is presented in Table 1 (table 1). The essential oil, which has been subjected to prolonged exposure to temperature, contains mainly oxidized, acetylated and aromatic structures. In essential oil the effect of temperature has been minimized. Therefore, there are more reduced forms, OH groups, and unsaturated compounds. It is very important to note that the essential oil obtained by the second method contains provitamin A and a derivative of vitamin D.

Table (1):The main compounds identified in the essential oil of tansy grass obtained by two methods (the essential oil content in the sample is 1.00 ± 0.11%).

S. No	Name	Formula	Rt	Essential oil obtained by the Ginsberg method ,%	Essential oil obtained by the Clevenger method,%
1.	o-Cymene		8.722	0.40	0.24
2.	Eucalyptol		8.847	3.42	5.43
3.	Artemisia ketone		9.478	0.94	1.8
4.	Terpineol,		9.570	0.02	0.09
5.	Tanacetone		10.300	4.98	5.21
6.	.(+)-Camphor		11.030	15.97	16.50
7.	Isoborneol		11.431	12.48	13.16
8.	4-Terpineol		11.582	3.52	3.48
9.	.Karvakrol		13.502	0.80	0.49
10.	CaryophylleneCaryophyllene oxide		15.212	2.80	1.78
11.	Limonen-6-ol, pivalate		16.967	1.29	1.13
12.	(-)-Spathulenol		17.303	1.06	1.22
13.	à-Cadinol		17.671	0.86	1.60
14.	á-Eudesmol		18.197	3.38	2.13

Table (2):Compounds identified in tincture based on tansy grass.

No.	Name of the compound	Formula	Rt	S	%
1	(+)-Camphora		11.030	2435899	2.4
2	Ascaridole epoxide		18.140	27736	0.1
3	(-)-Myrtenyl acetate		18.847	1100370	1.0
4	D-Limonene		19.192	1052558	0.9
5	Cubenol		19.853	338868	0.3
6	Caryophyllene		20.699	1002562	0.9
7	á-Cubebene		21.806	2214492	2.0
8	7-Ethynyl-1,4a-dimethyl-4a,5,6,7,8,8a-hexahydro-2(1H)-naphthalenone		22.437	126159	0.1
9	ç-Selinene		22.510	193507	0.1
10	Limonen-6-ol pivalate		23.049	2435595	2.2
11	Spathulenol		23.478	857776	0.7
12	Caryophyllene oxide		23.592	1941455	1.7
13	Транс-лонгипинокарвеол		25.246	1316935	1.2
14	Isoaromadendrene epoxide		27.164	827993	0.7
15	Achillin		32.648	1206402	1.1

When studying antioxidant activity, it was found that the maximum antioxidant activity is shown by petroleum extraction when heated to 40°C, petroleum extraction at normal temperature shows high antioxidant activity and when diluted with alcohol, antioxidant activity decreases approximately twice. The essential oil obtained by the first and second methods exhibits approximately the same antioxidant activity, although the antioxidant activity of the essential oil obtained by the second method is somewhat higher, which is apparently associated with a higher content of reduced compounds compared to the essential oil obtained by the first method. In the experiment, the antioxidant activity of the tincture is slightly inferior to other medicinal forms from medicinal plant raw materials of tansy grass, which makes it possible to recommend all of the above extracts for further study of pharmacological activity. Essential oil obtained from tansy grass by the first and second methods, tincture obtained by maceration and petroleum extraction from tansy grass showed high antioxidant activity,С, mg/g×10(figure 2).

Figure (2): Antioxidant activity of extracts from tansy grass.

DISCUSSION:

At the moment, oil extracts from tansy grass have not been studied at all, where mineral oil - petroleum is used as an extractant. Petroleum is a fraction obtained by rectification of oil in the temperature range from 200 - 300°C Petroleum is used as a drug with an anti-inflammatory effect. Petroleum is a multicomponent extractant in which more than 200 compounds have been identified by chromatography-mass spectrometry. The pharmacological activity of petroleum extracts from various types of medicinal plant raw materials is studied: from peppermint leaves, from medicinal sage leaves, from walnut fruits, from pumpkin seeds. These extracts in the experiment show high antiparasitic activity. It is advisable to study the pharmacological activity of petroleum extraction from tansy grass. In preliminary experiments, this drug showed high antiparasitic activity. To conduct detailed preclinical studies, it is necessary to develop methods of standardization of both medicinal plant raw materials of tansy grass and petroleum extraction from tansy grass. Based on the literature data, there are three main classes of compounds with high pharmacological activity in the common tansy plant. Essential oils, which are associated with anthelmintic properties, flavonoids and phenolic carboxylic acids, with which hepatoprotective properties are correlated. In petroleum extraction, it was possible to identify, in addition to volatile compounds characteristic of oil, essential oil components, including camphor, according to the content of which end-to-end standardization of medicinal plant raw materials of common tansy grass, essential oil, tincture and petroleum extraction is possible.

As a result, it was concluded that:

1. The method of chromatography-mass spectrometry revealed the multicomponence of drugs obtained from medicinal plant raw materials of Tanacetum vulgare. In essential oil, regardless of and modification of the method of preparation (hydrodistillation using a Ginsberg method or hydrodistillation using a Clevegermethod, more than 80 compounds have been identified; more than 100 compounds have been identified in the tincture, in petroleum extraction more than 150 compounds from which marker compounds such as camphor, borneol, ascaridiol, isoaromadendrene, cubenol, limonene, carvacrol, pinene, cineol, terpineol and chrysanthenone can be isolated). Comparative analysis of essential oil samples obtained by different methods showed that mainly oxidized, acetylated and aromatic structures are present in essential oil that has been subjected to prolonged exposure to temperature (Ginsberg method). In essential oil, where the effect of temperature was minimized (the Clevenger method), there are more reduced forms, OH groups, and unsaturated compounds. The essential oil obtained by the second method contains provitamin A and a derivative of vitamin D.

2. The wider component composition of the tincture obtained from tansy grass compared with essential oil is explained by the presence of flavonoids, organic acids, tannins and a more diverse composition of vitamins in the alcohol extraction. Vitamin E and provitamin A have been identified.

3. The maximum amount of biologically active compounds was identified in the petroleum extraction from tansy grass, due to the multicomponent composition of the extractant- petroleum. This dosage form is promising for further study of anthelmintic activity, where there is a synergistic effect of the extractant and extract from medicinal plant raw materials.

4. The maximum antioxidant activity is shown by the petroleum extraction from the tansy grass. Tincture and essential oil exhibit antioxidant activity slightly lower than petroleum extraction.

CONFLICT OF INTEREST:

The authors declare no conflictof interest.

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Received on 17.03.2022 Modified on 19.07.2022

Research J. Pharm. and Tech 2023; 16(1):229-235.

DOI: 10.52711/0974-360X.2023.00042