A TLC-densitometric method for determination of allantoin in Symphytum officinale root was developed. Densitometric quantification of allantoin was carried out on TLC Si60 plates with butanol-50 % methanol/formic acid, 66.5:33.2:0.3 (V/V/V) as developing solvent, at a wavelength of 190 nm. The method was preliminarily validated in terms of specificity, linearity, precision, limit of detection, limit of quantification, recovery and robustness. The results of TLC quantification were compared with HPLC analysis carried out on a HILIC Luna NH 2 100A column, with mobile phase consisting of acetonitrile/water 80:20 (V/V) and UV detection at 190 and 210 nm. Allantoin content was determined in two herbal products and it varied from 0.94 to 2.09 %, depending on the producer, and was in agreement with literature reports.

Context Trigonella foenum-graecum L. (Fabaceae) has many therapeutic properties and anticancer potential. Objective The cytotoxic activities of standardized extracts and a fraction from fenugreek seeds and their compounds (sapogenins, flavone C-glycosides, alkaloid trigonelline) against human cancer SKOV-3, HeLa and MOLT-4 cells were evaluated. Materials and methods Fenugreek seeds were extracted with 70% methanol (A) or water (B). Furthermore, the seeds were purified with petroleum ether and chloroform and next extracted with methanol to obtain fraction (C). The quantitative analysis of saponins and flavonoids in the extracts was done with HPLC methods. The extracts (5–120 µg/mL) and compounds (1–50 µg/mL) were tested on the cells by MTT assay and RTCA system. The effect of a fraction on ROS production, mitochondrial membrane potential and caspase-3/7 activity in HeLa and SKOV-3 cells was also evaluated by flow cytometry. Results The strongest cytotoxic activity on cancer cells showed the fraction C (IC 50 was 3.91 ± 0.03 for HeLa, 3.97 ± 0.07 for SKOV-3, and 7.75 ± 0.37 for MOLT-4) with the highest content of steroidal saponins (163.18 ± 11.03 μg/mg) and flavone C-glycosides (820.18 ± 0.05 μg/mg). The fraction significantly increased ROS production (up to four times higher than in keratinocytes as control) and caspases activity in the cells. The examined flavonoids did not exhibit the cytotoxic activity in contrast to yamogenin, tigogenin, and diosgenin. Conclusions The obtained results complement the data on the cytotoxic activity of Foenugraeci Semen and synergistic effect of flavonoids and saponins complex contained in the plant.

Summary Introduction: Arnica flowers are used in pharmaceutical and cosmetic industry. According to EMA only endangered Arnica montana provides the medicinal plant material. However, some European countries also allow the use of A. chamissonis flowers, whose chemical composition is not known in detail. Objective: The aim of the study was to recognize and compare the chemical composition of A. montana and A. chamissonis flowers collected from plants cultivated in Poland. Methods: HPLC-DAD-ESI/MS analysis comprised phenolic acids, flavonoids and sesquiterpene lactones. Results: Thirty eight flavonoids and phenolic acids were recognized and quantified, with patuletin, 6-methoxykaempferol and quercetin present only in A. chamissonis flowers. Moreover, helenalin and acetyl-dihydrohelenalin were identified. Conclusion: A. montana and A. chamissonis flowers from plants grown in Poland possess similar composition of simple phenols and polyphenols, present in concentration slightly higher in A. montana. Helenalin and dihydrohelenalin esters identified in arnica flowers from various regions of Europe were not detected.

The herbal drugs obtained from Arnica montana flowers are known for their anti-inflammatory activity. However, there is no detailed data on chemical constituents of the whole Arnica plant. HPLC-DAD-ESI/MS analysis of the whole Arnica montana plant (Arnica TM), used in the preparation of allopathic and homeopathic medicinal products, has been conducted. The qualitative and quantitative analysis comprised phenolic acids, flavonoids and sesquiterpene lactones ñ helenalin and dihydrohelenalin esters identified earlier in Arnica flowers. The analyzed material showed to be a rich source of caffeoylquinic acid (CQA) derivatives, especially 3,5di-O-caffeoylquinic acid. According to the literature data, the anti-inflammatory activity of extracts from the whole Arnica montana plant can be due to CQA presence as dominating compounds.

Comfrey root – Symphyti radix has been present in medicine since ancient times. Nowadays, European Medicine Agency (EMA) restricts its only for external use, in short therapies not exceeding 10 days. The last decade has been characterized by the progress in research on the chemical composition and metabolism of biologically active compounds present in the root and leaves. The result is the presence of herbal medicinal products containing comfrey root extracts with reduced content or free of toxic pyrrolizidine alkaloids on the European market of herbal products. Moreover, the medicinal properties of leaves from another Symphytum species, namely Russian comfrey, are also in the use. The results of clinical trials indicate the beneficial effects of plant raw materials obtained from comfreys in musculoskeletal system disorders – comfrey root and skin inflammations – leaves of Russian comfrey. The paper presents the history of use, chemical composition, therapeutic effect and toxicity of comfrey, as well as characterizes clinical trials on products containing comfrey extracts from root and leaves.