Comparative chromato-mass spectrometric study of tinctures based on Echinacea purpurea (L.) Moench

An urgent problem today is the spread of secondary immunodeficiencies associated with increased stress, urbanization, and negative changes in the environment that lead to impaired immune function. Herbal remedies are milder, less likely to become addictive and have side effects, and are long-lasting. A promising plant with immunomodulatory action is Echinacea purpurea (L.) Moench, a member of the family Asteraceae. The main active ingredients of echinacea are phenolic compounds, carbohydrates, and alkylamides. The chemical composition of echinacea is also represented by flavonoids, tannins, saponins, higher fatty acids, amino acids, betaine, essential oil, macroand microelements (Se, Co, Ag, Mo, Zn, Mg, K, Na, Fe, etc.). Drugs have immunostimulatory, antioxidant, membrane-stabilizing effect, promote healing of wounds, burns, ulcers, are used in infectious and viral diseases, especially HF. They are traditionally used for furunculosis, septicemia (infection of the blood), pyorrhea, tonsillitis, especially for the treatment of boils, carbuncles, and abscesses. It is also important that in modern pharmaceutical science the leading place is occupied by research related to the introduction into medical practice of herbal medicines, the study of their chemical composition, standardization, development of optimal technologies for the manufacture of phytopreparations. That is why the objects of our research were tinctures based on Echinacea purpurea.

Актуальные вопросы фармацевтической и медицинской науки и практики. 2021. Т. 14, № 3(37). С. 275-281 Comparative chromato-mass spectrometric study of tinctures based on Echinacea purpurea (L.) Moench Today is the actual problem there is a spread of secondary immunodeficiencies, related to high-stress loads, urbanization, and negative changes in ecology, that lead to violations functioning immune system. These states inevitably need it to carry out immune correction. Use for this purpose synthetic drugs complicated a significant risk of developing unwanted reactions and usually low economic availability of such medicines for the population. In return, drugs, which are produced based on medicinal plants, are characteristic with softer action, less likely to get used to it and side effects, as well as the opportunity long enough application [1,2]. Therefore, in the practice of rehabilitation of persons with secondary immunodeficiency states, including as support for chemotherapy in cancer, it is advisable to use medicinal plants that have immunotropic effects [1,3,4].
It is also important that in modern pharmaceutical science the leading place is occupied with the researches connected with the introduction into medical practice of herbal medicines, studying of their chemical composition, standardization, development of optimum technologies of production of herbal preparations. That is why tinctures based on purple Echinacea have become the object of our research [11].

Aim
The purpose of the work is to study and comparative analysis of the component composition of tinctures based on purple Echinacea (Echinacea purpurea (L.) Moench) using gas chromatography (GC).

Materials and methods
Tinctures of rhizomes with purple Echinacea roots were used for the study: 1) experimental -manufactured according to an industrial, technological recipe (1:5, extractant -ethanol 70 %) from raw materials, which was harvested in July 2019 at the experimental site of the Zaporizhzhia State Medical University. The receipt of this tincture was carried out according to validated technology by the approved technological regulations. The main stages of production are reflected in the flow chart (Fig. 1); 2) control -finished products of LLC "Zhytomyr Pharmaceutical Factory", series 20319.
The tinctures were examined on an Agilent 7890B gas chromatograph with a 5977B mass spectrometry detector. Chromatography conditions: DB-5ms column, 30 m long, with an internal diameter of 250 μm and a phase thickness of 0.25 μm. The carrier gas (helium) velocity is 1.3 ml/min. Injection volume -0.5 µl. Flow split -1:5. The temperature of the sample input block is 265 °C. Thermostat temperature: programmable -70 °C (holding 1 min), up to 150 °C at a speed of 20 °C/min (holding 1 min), up to 270 °C at a speed of 20 °C/min (holding 4 min). A NIST14 mass spectra library was used to identify the components.

Discussion
The results of the study show that the qualitative and quantitative chemical composition of rhizomes with roots of purple Echinacea was characterized by complexity and variability. This fact complicates the standardization of herbal preparations and may affect their effectiveness and safety. ISSN         2. In the experimental tincture of rhizomes with roots of purple Echinacea identified 31 components, and in the control -23. The composition of the tinctures was characterized by complexity and variability. In the tinctures studied, 15 components were common, with only 2 components in quantitative terms.
3. The results of the study were of value for the further development and improvement of analytical regulatory documentation for raw materials and herbal preparations of purple Echinacea. They may also be the basis for the creation of new immunomodulatory substances based on the underground organs of Echinacea.
Prospects for further research. Tinctures based on Echinacea purpurea (L.) Moench can be considered as a basis for the creation of new substances with immunomodulatory activity based on the underground organs of echinacea.

Funding
The work is part of the joint integrated work of the Department of Pharmacognosy, Pharmacology and Botany of Zaporizhzhia State Medical University.