INVESTIGATION OF THE POSSIBILITY OF USING THE METHOD OF DIFFERENTIAL SPECTROPHOTOMETRY FOR THE ANALYSIS OF OVERLAPPING ABSORPTION BANDS OF TRITERPENE SAPONINS CHROMOPHORES AND THEIR QUANTITATIVE DETERMINATION IN AQUEOUS SOLUTIONS

A detailed analysis of the absorption spectra of mono- and bidesmoside triterpene glycosides was performed. It is suggested that the maximum in the region of 198–208 nm is attributed to the absorption of the double bond in the cyclohexene ring of the aglycone. The second less seen maximum at a wavelength of 280 nm is observed in the spectrum of saponin Quillaja Saponaria and is almost completely absent in the spectrum of saponin Mukorossi Sapindus. Identification of this maximum is extremely difficult, since its presence can be due to both the aldehyde group in the aglycone and the hydroxyl and carboxyl groups in the carbohydrate molecules. Using the method of differential spectrophotometry, a method for decomposing the UV spectrum of saponins into its constituent components is proposed and justified. A spectral analysis was performed, including the assignment of the absorption bands to the functional groups of the studied compounds. The possibility of estimating changes in the spectral properties of glycoside solutions with changes in the acidity of the medium is considered. The possibility of a bathochromic shift of the maximum absorption of the aglyconic part of saponin depending on the microenvironment (the structure of the carbohydrate part) and changes in the pH of the solution is established. It is shown that the bands at 274, 280.5 nm correspond to n-π* transitions of carbonyl and carboxyl groups and are determined only in the regions of high concentrations in Quillaja Saponaria saponin solutions. Based on the results obtained, calibration models for the quantitative determination of saponins in solutions are proposed. The regression analysis of the calibration equations is carried out, the main statistical indicators are calculated.

DEVELOPMENT OF A METHOD FOR QUANTITATIVE DETERMINATION OF THE AMOUNT OF FLAVONOIDS IN TRIPLEUROSPERMUM INODORUM’S FLOWERS

Chamomilla recutita is used in scientific medicine. Tripleurospermum inodorum (L.) Sch. Bip. is widely spread among possible impurities of Chamomilla recutita (L.) Rauschert. This plant is perspective for establishment into scientific medicine but it can’t change Chamomilla recutita (L.) Rauschert. Purpose of research: development of a method for the quantitative determination of the amount of flavonoids in terms of rutin in Tripleurospermum inodorum’s flowers using differential spectrophotometry. Materials and methods. The samples prepared in different parts of Russia were used as objects of research. (2017 – 2020). Spectral studies were carried out in the wavelength range of 350-430 nm with a step of 1 nm using a spectrophotometer SF-2000. Results. To determine the analytical wavelength, the UV spectra of alcohol extraction of Tripleurospermum inodorum’s flowers were studied. Maximum of absorption was noticed at wavelength 370 nm. The differential spectrum of the same extraction with an aluminum chloride solution of 2% in 96% alcohol has a maximum at a length of 410±2 nm, which coincides with the maximum of the Standard Sample (SS) of rutin. The largest number of flavonoids is extracted by 70% alcohol. The maximum optical density and the highest output of the number of flavonoids from the raw material is observed at a degree of grinding of 2 mm with a single extraction for 60 minutes. In the conditions of complexation, the optimal ratio of the volume of the test solution and aluminum chloride with a solution of 2% in 96% alcohol is the ratio of 1:1. The stability of the complex with an aluminum chloride solution of 2% in 96% alcohol is observed in 40 minutes after the start of the reaction and retains it for 30 minutes. Findings. The method is developed and the parameters of the quantitative determination of the amount of flavonoids in Tripleurospermum inodorum flores are determined in terms of rutin using differential spectrophotometry.

THE PREDICTION OF THE ANTIOXIDATIVE ACTIVITY OF GROSSULARIA RECLINATA LEAVES

At present, in order to expand the raw material base, modern phyto-production is most interested in the use of medicinal plants' organs that were not previously used, but containing flavonoids. The Grossularia reclinata leaves, which are a cheap and affordable plant material, can become a promising source for the creation of phytopreparations containing flavonoid compounds. The aim of the study was to identify and quantify the sum of flavonoids' substances in terms of rutin in the Grossularia reclinata leaves. Material and methods. Qualitative reactions were selected to confirm the presence of flavonoids' substances; differential spectrophotometry was used for their quantitative assessment. Results. It was found that the total content of flavonoids in terms of rutin in alcohol extract from these leaves is on average 0.557 ± 0.0061%, which exceeds their content fruits that were used in medicine by 1.86 times. Comparative analysis of the flavonoids' sum of in the medicinal plants' leaves made it possible to determine that this group of biologically active substances in the Grossularia reclinata leaves is represented in greater quantities than in the motherwort herb (0.2%), wormwood (0.3%) and the tripartite series (0 ,five%). Conclusions: The results of the study allow us to consider this plant raw material promising for the production of new therapeutic and prophylactic drugs.

Implications of Fagopyrin Formation In Vitro by UV Spectroscopic Analysis

The present work aims at studying the possible biosynthesis of fagopyrin in buckwheat plants with an attempt to address the existing gaps. The developed method of differential spectrophotometry can be used for identification of naphthodianthrones fagopyrins. It was found that in the vegetative mass of buckwheat plants, fagopyrin precursor-2-(piperidine-2-yl)-emodindianthron could be present. As fagopyrin can be produced by light effect, the temperature factor may influence the formation of protofagopyrin in vitro. An optimum temperature range was estimated for protofagopyrin formation. A possible fagopyrin biosynthesis under in vitro conditions was suggested.

DEVELOPMENT OF COMPOSITION, TECHNOLOGY AND ANALYSIS OF GRANULES WITH LARYNX TREE EXTRACT LIQUID

The purpose of the study was to substantiate the optimal composition, development of technology and methods for analyzing a new dosage form - granules with hydrangeas with a liquid tree extract. Optimal auxiliary substances for granule production were experimentally selected and process parameters were determined: fractional composition, abrasion, disintegration, mass loss during drying, bulk density. It was found that for all indicators granules with hydrangea tree extract liquid meet the requirements of GF XIV. Process diagram of granules manufacturing by wet granulation method has been developed. The TLC method made it possible to identify rutin, quercetin, luteolin, chlorophyll, coumarin. The method of differential spectrophotometry was tested for the method of quantitative determination of the sum of flavonoids. The proposed quantification methodology has been found to be suitable for analytical purposes. The content of the sum of flavonoids in the granules in terms of rutin is about 0,0225%. The relative error of the technique does not exceed ± 2,5%.

New approaches for the standardization of the Monarda fistulosa herb

Objectives. The Monarda fistulosa herb of the Lamiaceae family is particularly interesting among essential oils from medicinal plants that have a wide spectrum of pharmacological activities. However, information regarding some of its flavonoids, which are found in the essential oil, is controversial. Inaccuracies in identification of the chemical composition of the herb have led to several different standardization approaches, which are cumbersome. To establish a uniform classification, here, we present confirmation for new approaches for the standardization of the Monarda fistulosa herb. Methods. Silica gel column chromatography was used to extract the flavonoids. Identification was based on ultraviolet spectroscopy, nuclear magnetic resonance spectroscopy, mass spectrometry, and acidic hydrolysis. The quality of the proposed quantitation methodology for total flavonoids was assessed by differential spectrophotometry at 394 nm, in isorhoifolin equivalent.Results. We have verified new approaches for the standardization of the Monarda fistulosa herb. The approaches can determine the authenticity of the herb by detecting monoterpene phenols and flavonoids that have diagnostic value. We also developed a technique for quantitation of the total flavonoids. Conclusions. We investigated the possibility of establishing the authenticity of the Monarda fistulosa herb based on the diagnostically significant flavonoids, isorhoifolin and linarin.

Development of the quantitative test methods to determine the number of flavonoids in flowering shoots of semi-soft hawthorn

Flowers, fruits and shoots of various genus Hawthorn (Crataegus L.) species are used in domestic and foreign medical practice as cardiotonic agents. It is also possible to use hawthorn raw material for the prevention of cardiovascular system diseases among medical and pharmaceutical personnel. We consider semi-soft hawthorn Crataegus submollis Sarg. to be a prospective raw material species as its flowering shoots are chemically similar to the red-haw hawthorn flowers. We developed the quantitative test method to determine the number of flavonoids expressed as hyperoside in flowering shoots of semi-soft hawthorn using differential spectrophotometry with an analytical wavelength of 412 nm.

Development of the method for spectrophotometric quantitative determination of the total flavonoids in Bupleurum multinerve herb

The article presents the method for the quantitative determination of flavonoids for the analysis of Bupleurum multinerve herb based on the method of differential spectrophotometry. The optimal conditions for analysis have been determined. They include extractant 40% ethyl alcohol, ratio of raw materials and extractant 1 : 100, extraction time of 60 minutes in the boiling water bath, complexing agent of 1 ml of 2% aluminum chloride. The use of rutin as a standard has been experimentally validated, analytical wavelength is 412 nm. The relative error of the mean result (for n = 9) was 3.20%. Validation studies of the method have shown that it meets the criteria: linearity (r = 0.99988), correctness, specificity, and precision. The analytical range of the method is 8.6726.08 g/ml. The method is recommended for the inclusion into the new edition of the Pharmacopoeia Monograph for this type of plant material.