Semi-quantitative method for determining coumarin derivatives in objects of various origin

Introduction. Coumarin and its derivatives are biologically active substances (BAS) of plant origin that exhibit a range of pharmacological activities as well as toxicological effects. The adverse effects of coumarin derivative drugs, along with poisonings caused by the use of rodenticides, coumarin-containing plants, and other related factors, justify the relevance of developing a rapid analytical method for the detection of coumarin derivatives.

Aim. To develop a rapid semi-quantitative method for the determination of coumarin derivatives using paper chromatography, perform validation tests, and apply the method to plant-based samples.

Materials and methods. For the development of the method were used substances coumarin and sodium hydroxide, filter paper brand FS and UV-lamp (365 nm). The technique includes impregnating paper bars with NaOH solutions (5–30 %), sample preparation with water or ethanol extraction followed by filtration, and visual fluorescence detection. Validation was performed using standard samples of phenolic compounds (coumarin derivatives, hydroxycorium derivatives, benzoic acids and flavonoids).

Results and discussion. As a result of the studies, the optimal conditions of the technique were established: concentration of the impregnating solution of sodium hydroxide – 10 %, use of SF-mark paper and detection time of 20 seconds. It is shown that the method has selectivity to coumarin derivatives when performing a number of tests, which are reflected in the «decision tree». Confirmed the possibility of semi-quantitative determination with detection limit 1 · 10^–6 mg/ml and testing on synthetic and plant objects.

Conclusion. A rapid semi-quantitative method for the determination of coumarin derivatives has been developed, based on their alkaline hydrolysis on a paper substrate with subsequent fluorescence detection. The method is characterized by simplicity, high sensitivity (limit of detection 1 · 10^–6 mg/mL), and was successfully tested on medicinal products, plant materials, and other objects. The proposed methodology is recommended as an effective tool for preliminary screening in laboratory practice.

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