CORRELATION OF INDICATORS OF INDEPENDENT METHODS FOR DETERMINING THE ANTIOXIDANT ACTIVITY OF BIOFLAVONOIDS

Sapieva Ardak Onalbekovna, Candidate of Chemical Sciences, Associate Professor of General and Biological Chemistry

NJSC «Astana medical university»

(010000, Republic of Kazakhstan, Nur-Sultan, 49A Beibitshilik street, e-mail: ardaksapieva73@mail.ru);

Kazbekova Ainagul Talgatovna, Candidate of Medical Sciences, Head of the Department of monitoring of clinical activity and interaction with clinical databases

NJSC «Astana medical university»

(010000, Republic of Kazakhstan, Nur-Sultan, 49A Beibitshilik street, e-mail: kazbekova_mua@mail.ru);

Madiyeva Sharapat Abdiganievna, Master of Agricultural Chemistry and soil science, Senior Lecturer at the Department of General and Biological Chemistry

NJSC «Astana medical university»

(010000, Republic of Kazakhstan, Nur-Sultan, 49A Beibitshilik street, e-mail: sharapat.828486@gmail.com);

Kenzheshova Akniet Kazbekovna, 3rd course student of the Specialty Pharmacy

NJSC «Astana medical university»

(010000, Republic of Kazakhstan, Nur-Sultan, 49A Beibitshilik street, e-mail: vousmevoyez.2000@mail.ru);

Baysarov Gabiden Maratovich, Master of Technical Sciences, Head of the Laboratory of natural compounds chemistry

JSC " International research and production holding "Phytochemistry"

(010000, Republic of Kazakhstan, Karaganda, e-mail: info@phyto.kz);

Seytembetov Talgat Sultanovich, Head of the Department of General and Biological Chemistry

NJSC «Astana medical university»

(010000, Republic of Kazakhstan, Nur-Sultan, 49A Beibitshilik street, e-mail: stalgat49@mail.ru);

Adekenov Sergazy Mynzhasarovich, Doctor of Chemical Sciences, Professor, Chairman of the Board

JSC "International research and production holding "Phytochemistry"

(010000, Republic of Kazakhstan, Karaganda, e-mail: info@phyto.kz).

Kazakhstan has unique reserves of plants of wild-growing species with medicinal properties, a significant part of which is promising for studies of their chemical composition and biological activity. Medicinal plants are the raw material for phytopreparations with a diverse spectrum of pharmacological and therapeutic effects, which, as a rule, are not accompanied by undesirable side effects.

Currently, many antioxidant and hepatoprotective drugs used in clinical practice are synthetic and cause allergic reactions. This circumstance determines the relevance of “free radical pathology” in pharmacotherapy and prevention of diseases and the corresponding use of herbal products, the effect of which is caused by the synergistic action of such natural compounds as polyphenols, aminophenol acids, higher fatty acids and others. In connection with the above aspects, the urgent problem of the development and implementation of phytopreparations and industrial use of plant materials in production [1,2]. Effective therapy of chronic liver diseases, as well as the prevention of drug hepatopathies are some of the urgent problems of modern medicine. In the pathogenesis of liver damage, oxidative stress is of great importance. One of the factors modifying the phospholipid composition of biological membranes is the development of free radical oxidation of unsaturated fatty acids in them. Flavonoids have the ability to neutralize reactive oxygen species and terminate chain reactions; therefore, a wide range of biological activity and their low toxicity, the absence of allergenic and cumulative properties allow us to consider bioflavonoids as a promising source for practical use in medicine [3,4].

The method for determining the iron-reducing potential is FRAP (FerricReducingAntioxidantPower) of the studied compounds. To 0.1 ml of an alcohol solution of the test sample in the concentration range of 0.25; 0.5; 0.75 and 1.0 mg / ml are added 0.25 ml of phosphate buffer (0.2 M, pH 6.6) and 0.25 ml of 1% potassium hexacyanoferrate (III) solution. The reaction mixture is incubated for 20 minutes at 50 ° C, the reaction is stopped by the addition of 0.25 ml of a 10% solution of trichloroacetic acid. The mixture is centrifuged for 10 minutes at 3000 rpm. The upper layer of the resulting solution with a volume of 0.5 ml is mixed with 0.5 ml of distilled water and 0.1 ml of 0.1% FeCl₃. The absorbance is measured at 700 nm with an AgilentCary 60 spectrophotometer. Ascorbic and gallic acids are used to evaluate antioxidant activity (AOA).

Determination of antiradical activity (ARA) by inhibition of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical by the analytes. 0.1 ml aliquots of the test sample in the concentration range of 0.01-1 mg / ml were added to 3 ml of 6 × 10- 5M ethanol solution of DPPH radical. After vigorous stirring, the solutions were left in the dark for 30 minutes. The optical density was measured on an AgilentCary 60 spectrofluorometer at 520 nm, butylhydroxyanisole was the standard [5].

The use of the method of initiated chemiluminescence (CL) to determine the antioxidant and antiradical activity. The intensity and dynamics of chemiluminescence were recorded on a CaryEclipce spectrofluorometer (Agilent Technologies) at 420 nm. The method is based on the free radical oxidation of luminol with hydrogen peroxide (pH 7.4), which results in the emission of light, which decreases in the presence of a potential antioxidant [6,7].

In the aspect of studying the correlation between the data of different methods for assessing bioactivity, the issue of the relationship between the data of different indicators is relevant. For example, for antioxidant activity, we performed a comparison between the compounds of the optical density, concentration and antioxidant effect values ​​(%) of a substance. It has been established that there is a definite correlation between these indicators: an increase in antioxidant activity with an increase in concentration and an increase in antioxidant activity with an increase in the optical density of a solution. The obtained results indicate the presence of a single mechanism of inhibition of peroxide processes in the presence of exogenous flavonoids. In this work, we evaluated the antioxidant activity of flavonoids dihydroquercetin (DHQ); 2- (3,4-dimethoxyphenyl) -5-hydroxy-3,6,7-trimethoxy-4H-chromen-4-one (Af-1); 5-hydroxy, 7-methoxy-2-phenyl-4H-chromen-4-one (Pb-3) and (R, E) -7-methoxy-4- (propan-2-ylidene hydrazine)-2-phenyl chromen-5-ola (G3Pb). The antioxidant property of flavonoids pinostrobin and oxymapinostrobin was established, which was compared with the similar effect of ionol, butylhydroxyanisole and other antioxidants. An indicator of the antioxidant activity of the studied objects is the optical density, in particular, AK has the highest optical density, and for compounds Af-1, Pb-3, and G3Pb, the antioxidant effect is less pronounced.

Flavonoids dihydroquercetin (DHQ) and oximpinostrobin, using the example of luminolinated chemiluminescence, inhibited free radical oxidation and the degree of effect depended on the concentration of exogenous substance, which follows from the form of the kinetics curve of this reaction (Figure 2) [8, 9]. An analysis of the literature on chemiluminescent analysis of both individual organic compounds isolated from plants and subsequent chemical modification, and total objects (plant extracts) indicates the prospects of this method and may find application in further studies on the development of new antioxidants and hepatoprotectors [10].

Concept and design: A.O. Sapieva, A.T. Kazbekova, Sh.A. Madiyeva.

Data collection and processing: A.K. Kenzheshova, Sh.A. Madiyeva.

Provision of research materials: G.M. Baysarov, S.M. Adekenov.

Analysis and interpretation of data: A.O. Sapieva, T.S. Seytembetov.

Manuscript Preparation: A.O. Sapieva, A.T. Kazbekova, Sh.A. Madiyeva, A.K. Kenzheshov.

Final approval of the manuscript: A.O. Sapieva, T.S. Seytembetov.

Administrative support: A.O. Sapieva.

The study did not have sponsorship.