Abstract
The aim of this work was to improve the approaches for standardization of Thymus L. species by development of HPTLC identification of phenolic compounds (flavonoids and phenylpropanoids) and terpenoids (essential oils and triterpenoids) and quantitative determination of rosmarinic acid and flavonoids for six Thymus L. species of Ukraine flora, as well as determination of chromatographic profiles of Thymus L. species’ extracts obtained using different extraction methods.
 Materials and methods. Objects were: samples of dry herb of Th.serpyllum, Th.vulgaris, Th.Marschallianus, Th.Pallasianus, Th.calcareus, and Th.moldavicus of Ukraine origin collected in 2019-2021 years or acquired commercially on Ukraine market. Instruments: CAMAG HPTLC Herbal System, visionCats 2.5. The analytical grade reagents were used. Reference substances were purchased from Extrasynthese, Sigma Aldrich. Chromatography was performed on HPTLC plates Si 60 F254, Merck according to the developed methods.
 Results. A new approach for quality control of different Thymus L. species of Ukraine flora included the development of HPTLC methods for identification of main groups of bioactive substances of these species, such as flavonoids and phenylpropanoids, essential oils, triterpenoids; development of quantification method of rosmarinic acid and assay of total flavonoids, expressed as luteolin-7-O-glucoside. The characteristic HPTLC fingerprints of six Thymus L. species in three mobile phases of different polarities that cover a wide range of bioactive substances were established. The content of rosmarinic acid in different Thymus L. species samples was in the range of 0,11-0,72 %: Th.moldavicus – 0,11 %; Th.Marschallianus – 0,19-0,27 %; Th.serpyllum – 0,38 %; Th.vulgaris – 0,51 %; Th.calcareus – 0,56 %; T.Pallasianus – 0,72 %. The total flavonoids content, expressed as luteolin-7-O-glucoside, was in the range of 0,8-2,72 %: Th.moldavicus – 0,8 %; Th.serpyllum – 0,87 %; Th.vulgaris – 1,06 %; Th.Pallasianus – 1,28 %; Th.Marschallianus – 1,89 %; Th.calcareus – 2,72 %.
 Conclusions. The proposed scientific approach for quality evaluation of Thymus L. species using HPTLC allows to determine comprehensive information of chemical composition and content of active substances of multiple samples in parallel, in a cost and time-efficient manner
Highlights
The Thymus L. genus belongs to the Lamiaceae family and includes several hundred species distributed in Eurasia, North Africa, Greenland [1,2,3,4]
A new approach for quality control of different Thymus L. species of Ukraine flora included the development of high-performance thin-layer chromatography (HPTLC) methods for identification of main groups of bioactive substances of these species, such as flavonoids and phenylpropanoids (ID 1), essential oils (ID 2), triterpenoids (ID 3); development of quantification method of rosmarinic acid (QD RA) and a specific assay of total flavonoids, expressed as luteolin-7-O-glucoside (TFC); as well as determination of specific HPTLC fingerprints for different Thymus L. species that are important for proper authentification and discrimination of potential falsification
The proposed scientific approach for quality evaluation of Thymus L. species using HPTLC allows to determining the comprehensive information of chemical composition and content of active substances of multiple samples of various species, obtained by a different technology, in parallel, in a time and cost-efficient manner
Summary
The Thymus L. genus belongs to the Lamiaceae family and includes several hundred species distributed in Eurasia, North Africa, Greenland [1,2,3,4]. The collectors of herbal raw material do not distinguish the species of Thymus, so its herbal raw material could be represented by different related species, as well as their combination in the various ratios [3, 4]. The modern high-performance thin-layer chromatography (HPTLC) method is included in the pharmacopoeial analysis [7]. It gives the possibility to compare results of different species or extraction in parallel at the same HPTLC plate or to compare tracks from different plates using “virtual plate”. That enables to use of the HPTLC method for comprehensive evaluation of the quality of closely related plant species
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