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THE USE OF THE DEVELOPED TLC METHOD FOR ANGIONORM COMPLEX HERBAL PREPARATION IDENTIFICATION BASED ON MARKER COMPOUNDS

DOI: https://doi.org/10.29296/25877313-2018-05-02
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Issue: 
5
Year: 
2018

P.A. Struchkov Post-graduate Student, Assistant, Chemistry Department, I.M. Sechenov First MSMU (Sechenovskiy University) (Moscow) V.L. Beloborodov Dr.Sc. (Pharm.), Professor, Chemistry Department, I.M. Sechenov First MSMU (Sechenovskiy University) (Moscow) Dr.Sc. (Med.), Chief Research Scientist, Department of Experimental and Clinical Pharmacology, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants (Moscow) I.V. Voskoboynikova Dr.Sc. (Pharm.), Leading Research Scientist, Department of Experimental and Clinical Pharmacology, All-Russian Scientific Research Institute of Medicinal and Aromatic Plants (Moscow) A.M. Savvateev Ph.D. (Pharm.), Assistant Professor, Chemistry Department, I.M. Sechenov First MSMU (Sechenovskiy University) (Moscow)

dry extract of the four herbs mixture: horse chestnut fruits (Aesculus hippocastanum L.), liquorice roots (Glycyrrhiza glabra L.), hawthorn fruits (Crataegi) and dog rose fruits (Rosae). The aim of the study was to develop Angionorm identification method by TLC based on previously selected marker compounds. The marker compounds of each raw material in the dry extract were previously determined by HPLC-MS-MS: glycyrrhizin for liquorice roots, β-escin for horse chestnut fruits, chlorogenic acid for hawthorn fruits and gallic acid for dog rose fruits. Different solvents and heating methods were tested for sample preparation optimization and the micro-wave heating with 80% ethanol (v/v) was chosen as the most suitable combination. The effective chromatographic separation was reached by using highperformance TLC plates (Merck Kieselgel 60 F254). The various organic solvents and acidic modifiers were tested as mobile phase components to optimize the separation. The most satisfying mobile phase was found to be water/formic acid/isopropanol/ethyl acetate/ toluene 10:10:35:30:15 (v). The marker compounds were detected by UV light (glycyrrhizin at 254 nm, chlorogenic acid at 366 nm) and by the detection reagent application. A number of reagents were tested; the best results were obtained by using vanillin/ sulfuric acid reagent, which allows detecting glycyrrhizin, escin and gallic acid. The developed method has optimal Rf parameters of the marker compounds, which are reliably extracted and easily detected. The method can be used for Angionorm tablets and dry extract identification
Keywords: 
TLC
Identification
Angionorm
horse chestnut fruits
liquorice roots
hawthorn fruits
dog rose fruits

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