Volume 8, (Spl-2- AABAS), December Issue - 2020, Pages:S327-S335 |
Authors: Alexander Loenidovich Mikhailov , Olga Arnoldovna Timofeeva , UlianaAleksandrovna Ogorodnova , Nikita Sergeevich Stepanov |
Abstract: The current study was carried out to study the effect of growing conditions on the pharmaceutically promising biologically active phenolic compounds on the meadow clover plants. The impact of the plant development site on the contemplated mixes was additionally explored. The estimation of phenolic compounds in raw plant materials was carried out with the help of a spectrophotometer and photocolorimeter. Results of the study revealed that the place of plant growth conditions have remarkable effect on the content of the phenolic compounds. Further, the most ideal conditions available for clover plant growth and higher phenolic compounds were reported from the broad-leaved forests. Furthermore, the results also suggested that the most important elements which affecting the aggregation of phenolic compounds are soil nitrogen. |
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Full Text: 1 Introduction The development of biologically active substances (BAS) in plants is a dynamic process and it is dependent on the plant ontogenesis and growing conditions (Jaakola & Hohtola, 2010). Plants of the same species grown under various natural conditions have differences in the biosynthesis and collection of optional metabolites (Koricheva & Barton, 2012). Among the reported metabolites, phenolic compounds, alkaloids, saponins, and terpenes are some common ones. Further, in reported secondary metabolites, phenolic mixes might be the biggest group of plant metabolites. They are broad in plants and contribute essentially to the shading, taste, and fragrance of numerous plant spices, nourishments and drinks. Some of the phenolic intensifies are pharmacologically important for their various medicinal properties. Huge numbers of the phenolic atoms are additionally viable cell reinforcements and free extreme scroungers, particularly flavonoids. Clover (Trifolium) is a genus of the family Fabaceae that includes about 300 species. Red clover (Trifolium pratense L.) is a perennial or biennial herb with a well-developed tap root system with a large number of lateral and hypoctile adventitious roots. The leaves are tri compound, almost glabrous, the lower ones are long while the upper ones are short petioles. The leaflets of the lower leaves are obovate, the upper ones are oval or ovate, usually more pubescent below, the stipules are spurious, narrowed (Hoekstra et al., 2017). This plant has a long history of ethnomedicinal uses and various attempts have been taken to explore the phytochemical profile, physiological effects, and possible therapeutic action of various biochemicals (Kolodziejczyk?Czepas 2016). The objective of this study was to identify the effect of soil and climatic conditions on the accumulation of phenolic mixes in clover plants filling in the Republic of Tatarstan. 2 Materials and Methods The aboveground parts of red clover were used as objects of study. The clover plant material was sampled from various "model" populations viz., deciduous forests (Kamsko-Ustinsky district, Verkhne-Uslonsky district, Apastovsky district, Tetyushsky district, Laishevsky district); coniferous-deciduous forests (Zelenodolsk district); southern taiga (Atninsky district); forest-steppe zone (Spassky district) of the districts of the Republic of Tatarstan: The assay of phenolic compounds in raw plant material was carried out by spectrophotometry and photocolorimetry. Distilled water (1.5 ml) was added to the 50 mg raw material and boiled it 45 minutes in a water bath at a temperature of 70°C. After this, the mixture was centrifuged at 15,000 rpm, and the resulted mixture was used for further study. From the prepared aqueous extracts, 75 μl of the aqueous extract was taken in the 1.5 ml vial and 75 μl of Folin-Denis reagent was added to it, and the mixture was stirred for three minutes, this was followed by the addition of 120 μl NaCHO3 solutions (10%) and 1.2 ml of water. The prepared mixture was left for 45 minutes and after this centrifuged at 16 thousand rpm for 2 minutes. Then, the optical density of the supernatant was determined at a wavelength of 725 nm. For a control solution, 75 μl of solvent was used instead of the extract. The total content of soluble phenolic compounds was calculated by using Sibgatullina (2011) formula: C = E*?*R*V / m*100 Where C - the concentration of phenolic compounds (mg/g dry weight of the sample);E - optical density at 725 nm; R dilution, times; V - volume of extract (ml); ? - reference substance (epicatechin (480) conversion factor); and m - sample weight of plant material (g). The assay of flavonoids in terms of quercetin, avicularin, and cenaroside was conducted as follows: 1 g thin section of the raw material analytical sample was placed in a 150 ml flask, this was followed by the addition of 30 ml of 90% alcohol containing 1% concentrated hydrochloric acid, the flagon was associated with a reflux condenser and warmed on bubbling water shower for 30 min. The jar was then cooled to room temperature and sifted through a paper channel into a 100 ml volumetric carafe. The extraction was rehashed again as mentioned above with 90% alcohol for 30 min and the filtrate was again diluted by adding 25 ml of 90% alcohol. From this solution, 2 ml was taken out in a 25 ml volumetric flask and 1 ml of 1% aluminum chloride solution prepared in 95% alcohol was added and the volume of the solution was diluted up to the mark with 95% alcohol (Klyshev et al., 1978). The content of the total flavonoids cynaroside, avicularin, and quercetin in absolutely dry raw materials was calculated in percent as per the given formula. |
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