Abstract
The analytical task of determining the phenolic compound content of water-ethanol extracts of Populus tremula L. (common aspen) leaves is complicated by the heterogeneity of compound groups having different polarities and appearing in varying concentrations. The purpose of the present work is to study the conditions of solid-phase extraction and high-performance liquid chromatography used to analyse the content of different groups of phenolic compounds in water-ethanol extracts of leaves from the P. tremula plant. In order to facilitate the derivation of phenolic compounds, an exhaustive extraction process was carried out using ethanol. Solid-phase extraction was carried out using a Diapak C16 cartridge, after which the eluates were passed through a membrane filter having a pore diameter of 0.45 μm. The high-performance liquid chromatography method was used to determine the content of phenolic acids and flavonoid glycosides, as well as salicin and individual flavonoid glycoside components: hyperoside, rutin, astragalin and two unidentified flavonoid glycosides in aqueous (analyte 1) and aqueous-alcoholic fractions (analyte 2) in two systems along the gradient elution. The requirement of analysing the primary aqueous eluate together or in parallel with the main aqueous-alcoholic fraction in the preparation of P. tremula leaf extracts for high-performance liquid chromatography using solid-phase extraction cartridges was substantiated. For separating the extract to determine the hydroxycinnamic and hydroxybenzoic acid content, it is preferable to use system 2; for determining the phenologlycoside (salicin) content, system 1 is more effective. Flavonoid glycosides (hyperoside, rutin, astragalin and two unidentified flavonoids) make the most significant contribution to the difference between the aqueous and aqueous-alcoholic fractions.
Highlights
The study of phenolic compounds (PCs) in plant material is of current interest in order to identify new economically-significant sources of biologicallyactive substances, primarily for medicinal use
MATERIALS AND METHODS We examined mature undamaged leaf blades from aspen trees of between 10 and 15 years old, collected from the 1st to the 5th August, 2015, on the territory of the Kedrovsky coal mine in southwestern Siberia
For solid-phase extraction (SPE), 1 ml of the extract was diluted with bidistilled water to 5 ml and passed through a Diapak C16 concentrating cartridge (CJSC BioKhimMak ST)
Summary
The study of phenolic compounds (PCs) in plant material is of current interest in order to identify new economically-significant sources of biologicallyactive substances, primarily for medicinal use. For taxonomic and bioindication purposes, it is necessary to obtain the most complete information on the content of individual phenolic substances, which presupposes exhaustive extraction and careful sample preparation. In terms of significantly simplifying the analysis procedure and improving its metrological characteristics, the most efficient and versatile method for isolation, purification and concentration of phenolic substances from plant samples having a complex composition is solid-phase extraction (SPE) [2, 3]. Mention of the use of solid-phase extraction (SPE), including the use of cartridges such as Phenomenex Strata-X and C18 (Torrance, CA, USA), Agilent SampliQ (Agilent Technologies, CA, USA) separately or together with other methods of purification and / or fractionation, was found in about a third of the reviewed publications [4–13]. A sorbent with a grafted C18 phase is optimally used for sample preparation within the framework of HPLC analysis [1]
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