STUDY OF FLAVONOID AGLICONES OF THE MULBERRY WHITE LEAVES DRY EXTRACT BY THE HPLC METHOD
DOI:
https://doi.org/10.11603/mcch.2410-681X.2020.i4.11744Keywords:
white mulberry leaves, dry extract, aglycones, quercetin, kaempferol, myricetinAbstract
Introduction. The flavonoids of the white mulberry leaves are represented by various glycosides, which complicate the standardization of their content of both – raw materials and its extracts. The spectrophotometric determination of flavonoids is difficult due to the variety of glycosides forms of flavonoids, and the presence of other phenolic nature substances, which have the similar to flavonoids analytical properties.
The aim of the study – to research the composition and content of flavonoid aglycones in the white mulberry leaves dry extract.
Research Methods. Dry extract of the white mulberry leaves was obtained from raw materials of domestic origin by fractional maceration; extractant – ethanol (80–60 %, v/v); extraction ratio – 1: 9-15, the multiplicity of extraction – 5, the currency of the one extraction – 24 h. Laborota 4011 rotary evaporator (Heidolph Instruments GmbH & Co. KG, Germany), SP-100 drying cabinet (UOSLAB, Ukraine), Mettler Toledo XP205DR analytical scale (Mettler Toledo, Switzerland), Bandelin Sanorex digitec electronic ultrasonic bath (GmbH & Co. KG, Germany), Agilent 1200 liquid chromatograph with G1315D DAD Detector (Agilent, USA), Kromasil 100 C18 (0.125 m × 4.6 mm, 5 μm, Supelko, USA) chromatographic column, aglycones standards (Sigma-Aldrich), chemical reagents of analytical grade and solvents of HPLC-grade (Sigma-Aldrich, Macron Fine Chemicals ™) were used in the study.
Results and Discussion. The flavonoid aglycones were obtained by the acid hydrolysis of the dry extract sample carried out during 2 hours. Separation of the obtained aglycones was performed by exhaustive extraction with ethyl acetate. For HPLC, the solvent was distilled off, and the dry residue containing the test aglycones was dissolved in methanol to prepare the test solutions. A number of peaks were observed on the chromatograms of the test solutions, three of which corresponded to myricetin, quercetin and kaempferol. Therefore, the flavonoids of the dry extract of white mulberry leaves are represented by glycosides of these three flavonols. Data on the presence of quercetin and kaempferol coincide with data from studies by other authors.
Conclusions. The composition of flavonoid aglycones of the white mulberry leaves dry extract was studied. It was found that they are represented by flavonols: quercetin, kaempferol and myricetin. 0.20 % of quercetin, 0.14 % of kaempferol and trace amounts of myricetin – 0.003 % were found. The obtained data can be used to develop a method of standardization of white mulberry leaves dry extract on the indicator "Quantitative determination".
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