The study of the phenolic substances of the aerial part of the plant "Kokamaron" ( Scutellaria Leptosiphon Juz. )

. In the article, the extraction of secondary metabolites, i.e., flavonoids, of the Scutellaria leptosiphon Juz plant, belonging to the Scutellaria family growing in Uzbekistan, and their chemical structure was studied. The methods of extraction, fractionation, thin-layer chromatography, column chromatography, paper chromatography, and quality reaction methods were used during the research. Also, the structure of flavonoids was analyzed by physical methods - ultraviolet (UV), infrared (IR), nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry. The aerial part of Scutellaria leptosiphon Juz collected from the mountains of the Sherobod district of Surkhandarya region (Uzbekistan) was studied for the first time and it was found that it contains baicalein, wogonin and baicalein-7-О -  -D-glucopyranoside. Based on the results of chemical reactions and UV, IR, 1N NMR spectrum data, the structure of 3 flavonoid substances isolated from the examined plant was determined to be Baicalein (5,6,7-trihydroxyflavone), Vogonin (5,7-dihydroxy-8-methoxyflavone) and Baicalein-7-О -  -D-glucopyronazide substances. The results and conclusions of the research conducted on the isolation and chemical structure of the secondary metabolites of the S. Leptosiphon plant belonging to the Scutellaria family growing in the flora of Uzbekistan were presented.


Introduction 1.Relevance of the topic
Over 6,000 plant species play a vital role in our daily lives.Around 1,500 of them have significant medicinal properties.Currently, extensive research is underway to extract medicinal compounds from plants, investigate their chemical composition and biological functions, and incorporate them into the national economy.The demand for natural remedies derived from plants is growing in modern medicine.While synthetic drugs can take effect quickly and efficiently, their long-term use can trigger harmful changes in the body.As a result, it is crucial to extract physiologically active compounds from plants, synthesize new derivatives, and develop effective medications to address this issue [1].
Plants are a limitless source of natural compounds containing various chemical structures and high biological activity.
Currently, natural remedies used in disease treatment have nearly no harmful effects on the body.
The structure of a plant organism is highly intricate, with a composition of multiple organic and mineral compounds.Many of these compounds can cure specific diseases, providing a healing effect.Plant organisms are the only ones that can synthesize organic compounds required to sustain their life from inorganic substances.Normally, organic compounds synthesized in plant tissue can be classified into two groups: Around the world, there are over 420 plant species belonging to the Scutellaria family (known as blueberry (Kokamaron) in the Russian -shlemnik family Lamiaceae), some of which are utilized in scientific and folk medicine.A considerable number of Scutellaria L. plant species grow natively in mountainous and sub-mountainous regions of Uzbekistan.Scutellaria L. plant-derived tinctures (extracts) are commonly employed in folk medicine for their antibacterial, antimetastatic, refreshing, blood pressure-modulating, and heartimproving properties, amongst many others.Tibetan medicine includes more than 40 Scutellaria L. plant species in phyto-infusion [2].Phenolic substances present in Scutellaria L. plants can account for their biological activity.
In both folk and scientific medicine, isolating and scientifically researching phenolic substances present in plant raw materials is of significant practical importance.Flavonoids abundant in plants of the Scutellaria L. genus exhibit high physiological activity, making them popular in medicinal preparations [3].
Since the chemical composition of the S. leptosiphon plant under investigation is insufficiently studied, researching flavonoids from these species growing within our nation's borders is an urgent and priority task.

Literature review
Over 65 Scutellaria plant species have been studied globally for their chemical composition, leading to over 330 isolated phenolic compounds.To date, foreign scholars, including Y. Imoto, H. Kizu, T. Namba, N. Joshee, Y. Y. Zhang, C. R. Yang, Z. H. Zhoi, J. Miao, T. Tomimori, S. Shibata, Y. Kikuchi, Y. Miaichi, I. I. Chemesova, N. K. Chirikova, V. I. Litvinenko, and T. P. have studied these plants' chemical structures.Popova, M. Linuma, and A. L. Budantsev have conducted research on the flavonoid chemical structure of these plants, identifying their pharmacological properties and using them as a basis for new, effective medical formulations.Substances such as baicalin, baicalein, and wogonin extracted from Scutellaria plants have antiviral and antibacterial properties; these substances have also emerged as an effective anti-inflammatory, anti-AIDS, anti-cancer, and anti-seizure agents.
In Uzbekistan, scientists such as V.M. Malikov, E.Kh.Botirov, Sh.V. Abdullaev, M.P. Yuldashev, R. Muradov, and F.D. Eshbakov have conducted research in this area.These scientists have isolated both new and known flavonoids from plants belonging to the Scutellaria L. family.The structure of the obtained substances was scientifically proven, and their pharmacological activity was determined.
These scientists have isolated several new and known flavonoids from plants belonging to the Scutellaria L. family and scientifically proven the obtained substances' structure and pharmacological activity.Based on this, the investigation of new species in the Scutellaria L. family is an urgent scientific and practical topic.
Many flavonoid substances have been extracted from studied Scutellaria (blueberry) plants.The vast reserve of plants in this group, along with many unexplored species, offers an excellent opportunity for isolating new and effective biologically active compounds, resulting in scientific and practical significance.These facts serve as a basis for selecting this study topic.

Research goals and objectives
The purpose of the work is to extract flavonoids from the S. Leptosiphon plant species belonging to the Scutellaria family collected from the slopes of Kugitang Mountain, Sherabad District, Surkhandarya Region (Uzbekistan) in July 2021, and to determine their chemical structure.
-S. Leptosiphon Juz.extraction of plant tops, separation into fractions, chromatography and other methods to extract pure substances; -Research of the chemical structure and properties of isolated flavonoids using chemical, instrumental and spectroscopic methods; -to determine the pharmacological activity of isolated flavonoids in our future scientific research; -Based on the sum of flavonoids of this plant, in our future scientific work, we will obtain natural dyes, and dye natural and artificial fabrics under their influence.

The scientific novelty of the research
Flavanoids in the aerial parts of Scutellaria Leptosiphon, a member of the Scutellaria plant, were studied.For the study of flavonoids, the plant was extracted with ethyl alcohol and fractionated with chloroform, ethyl acetate and butanol.Fractions were separated and studied in paper chromatography and thin-layer chromatography.Flavonoids were isolated by elution in different solvents in column chromatography columns.

The scientific and practical significance of research
With the creation of synthetic and chemical methods of obtaining medicine, various new compounds that do not occur in nature, including medicinal substances, began to be synthesized.In general, there have been many cases of negative effects on the immune system of the human body.Proper use of medicinal plants in the territory of our republic, identification, isolation, analysis of physiologically active substances contained in them, and the study of their pharmacological effects are of great importance.

Materials and methods
Extraction, fractionation, thin-layer (TLC), column (CC) and paper (PC) chromatography methods and qualitative reactions were used in the course of research.[4,5].Various colour reactions were used to determine the position of hydroxyl groups in the structure of flavonoids in paper and thin-layer chromatograms [6].

Flavonoid analysis reactions
1.When flavones, flavanones, flavonols, and flavanonols interacted with ammonia and alkali solutions, a yellow colour was formed.When heated, this colour changed to orange or red.
Flavanone and chalcones were analyzed with this reaction because flavanones are converted to chalcones under the influence of alkali [7][8][9].2. If Dimrot's reagent (a saturated solution of boric acid in acetic anhydride) is applied to the plant extract at 100-110 ℃, the yellow, orange colouring of the solution indicates the presence of 5-oxyflavonols and their methyl esters.When a 3-5% aqueous solution of boric acid is applied to the plant extract at room temperature, all flavonoids and other polyphenols with a dioxy group form a precipitate or yellow precipitate [10].
When a solution of boric acid in acetone is exposed to the plant extract at room temperature, the yellow colouration of the solution indicates the presence of 5-oxyflavones.At this time, flavonoids form complex compounds with boric acid: Physical methods: ultraviolet light (UV) [10,11].13S NMR spectroscopy [12].Mass spectrometry [11][12][13] methods were used.
For research, above-ground and root parts of Scutellaria leptosiphon Nevski, collected in July, were taken from a mountain slope near Kugitang Shark, Sherabad District, Surkhandarya Region of the Republic of Uzbekistan.UV-spectra of the studied substances were recorded on a Lambda-16 spectrophotometer (Perkin Elmer, USA) in solutions in ethanol and methanol, IR-spectra were recorded on a System 2000 FT spectrophotometer (Perkin Elmer, USA) in KBr-based tablets and solutions in glycerol, mass spectra were recorded on MX-1310 (Russia) obtained in a spectrometer at an ionization voltage of 50 eV.1N-YMR spectrum Tesla BS-567A (Czechoslovakia), 100 MHz (, m.b O-GMDS) in spectroscopy, 13S-YMR spectrum was obtained on the same device at 25 MHz.Chemical shifts are given in -scale in parts per million, and SSO'TD in -hertz.
The liquidus temperature was measured on a "BOETIUS" type viewing device.Column chromatography on KSK brand 100/160 μm silica gel (Reakhim-Russia, Chemapol-Czechoslovakia), thin-layer chromatography (TLC) on Silufol UV-254 plates and silica gel L 5/40 (Czechoslovakia) sorbent glass plate, paper chromatography (PC) Filtrak no.11 paper (Germany).In TLC, 1% AlCl3 solution in alcohol, 1% vanillin solution in concentrated sulfuric acid and 3% FeCl3 solution in alcohol, in the presence of ammonia vapour, and in paper chromatography by heating sugar substances with acidic aniline phthalate at 90-100 ℃ for 3-5 minutes identified with For the separation of flavonoids into individual compounds, the following mixture of solvents was used in column chromatography with ethylacetate and n-butanol compounds: 1. chloroform-methanol 97:3, 19:1, 9:1, 85:15, 4:1; 2. chloroform-isopropyl alcohol 97:3, 19:1, 9:1, 85:15, 4:1; The solvent system used in column chromatography for chloroform summation: Solvent system for thin layer chromatography (TLC).1. n-butanol-pyridine-water 6:4:3; 2. benzene-ethylacetate-acetic acid 50:50:1; The following reagents are used as bleaching reagents for chromatograms: Ammonia and iodine vapours, 50% potassium hydroxide solution dissolved in 10% ethanol, 2% zirconyl chloride solution dissolved in methanol.The condensed extract was diluted with water (1:1) in a separatory funnel, and the water-alcohol extract was extracted 6 times with gasoline from 0.400 l, and it was cleaned from oils and other additives.The purified extract was successively extracted 6 times with 400 ml of chloroform, 8 times with 350 ml of ethyl acetate, and 10 times with 350 ml of nbutanol in a separatory funnel and divided into fractions.Solvents were removed from the fractions and sums of fractions containing 10.0 g of gasoline, 14 g of chloroform, 21.0 g of ethyl acetate, and 26.0 g of n-butanol were obtained.

Separation of ethyl acetate compound
20.0 g of chloroform aggregate was separated into components using 1.8 x 140 cm silica gel sorbent 1.8 x 140 cm using 1-6 columns of chloroform: methanol system.The aggregate from the column was first washed with pure extracted chloroform and then with