DETERMINATION OF TOTAL FLAVONOIDS CONTENT IN CALENDULA TICNCTURE BY HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY
DOI:
https://doi.org/10.11603/2312-0967.2021.2.12182Keywords:
calendula tincture, high performance thin-layer chromatography, total flavonoids content, rutin, quantitative determinationAbstract
The aim of the work was to develop a method for determination of the total flavonoids’ content (TFC) of Calendula tincture by high-performance thin-layer chromatography (HPTLC) that is specific, fast and economy.
Materials and Methods. The investigation was carried out by HPTLC technique in automated system of CAMAG (Switzerland), on the base of Scientific-Research Training Laboratory of Chemical and Technological Research of National University of Pharmacy (Kharkiv, Ukraine). The results were evaluated using visionCats software, CAMAG. As a reference standard rutin was used, as objects of study three samples of Calendula tincture of domestic manufactures were used.
Results and Discussion. During development of TFC method the optimal conditions of analysis were chosen, including: determination of characteristic peak profiles of Calendula tincture in the selected mobile phase at different wavelengths; the selection of reference standard for expression, optimization of steps sample preparation for work and reference solutions, derivatization and detection; determination of calculation’s method and calculation formula. Thus, the determination of TFC was carried out after compounds separation in the selected mobile phase and after derivatization with aluminium chloride reagent at a wavelength of 408 nm. For the calculation, the calibration method – plot by peak area was chosen. According to the developed HPTLC methods, the TFC, expressed as rutin, in samples of Calendula tincture was: for sample 1 – 0,26 ± 0,02%; for sample 2 – 0,14 ± 0,01%; for sample 3 – 0,26 ± 0,02%.
Conclusions. The developed method can be used as a specific, convenient and economical express method for determination of TFC in Calendula tincture.
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