An Estimation of the Antiviral Activity and Toxicity of Biologically Active Substances Obtained from the Raw Materials of Artemisia cina Berg. In Vitro and In Vivo.

Species of the genus Artemisia are well known for their use as ingredients in ancient medicine. The advantage of using plant extracts compared to individual pharmaceutical ingredients is the rate of adaptation of the pathogenic microorganisms to the drug. Due to the rapid development of multidrug-resistance in microorganisms in the field, it is essential to search for novel, effective drugs with low toxicity. Therefore, the purpose of this study was to isolate and study the biologically active substances obtained from various substances in the raw materials of Artemisia cina Berg. The identification of the main biologically active components was performed using the method of chromato-mass spectrometry. Moreover, the antiviral activity of several extracts was studied using the method of measuring limiting dilutions (the Reed-Mench method), with some modifications. For the first time, the biological activity of extracts from the raw material of Artemisia cina Berg. upon the SARS-CoV-2 virus was confirmed. All the obtained extracts exhibited nontoxic effects in animals, with an LD50 greater than 2 g/kg. Comprehensive toxicological analyses are also presented in the study, such as those of the biochemical parameters of urine after one day and one week of the extracts' administration in mice at a dose of 2 g/kg body weight. In all groups of animals that received extracts of Artemisia cina Berg., a slight increase in the presence of red blood cells in their urine was observed one day following the administration of the extracts. This increase decreased somewhat after a week; however, it remained higher than the levels observed in the control animals. In the three groups, there was also a slight increase in the amount of ketones in the urine. Two weeks following the administration of the extracts to these groups, the internal organs of the animals were examined. The examination showed that the internal organs of the animals that received the extracts were not visibly different from those of the control animals in terms of their size or appearance. The weight of the internal organs of the animals that received the extracts was also similar to the weight of the internal organs of the control animals, illustrating the absence of toxicity.

Multidirectional Characterization of Phytochemical Profile and Health-Promoting Effects of Ziziphora bungeana Juz. Extracts.

Ziziphora species (Lamiaceae) have been used in traditional medicine as sedatives, antiseptics, carminatives, or expectorants. Despite their common applications in phytotherapy, there is still lack of evidence about the composition of their extracts and its impact on biological properties of the plants. The aim of this study was to evaluate the content of Ziziphora bungeana, a less studied species growing in Kazakhstan, using HPLC-ESI-QTOF-MS/MS instrumentation and to determine its antimicrobial, antioxidant, and cytotoxic activity together with inhibitory properties against tyrosinase and toxicity in erythrocyte lysis assay. Extracts from Z. bungeana were found to be sources of flavonoids, phenolic acids, organic acids, and terpenes that determined their antiradical activity. The minimum inhibitory concentrations of extracts were lower for Gram-positive bacteria (1.25-10 mg/mL) than for Gram-negative bacteria and fungi (5-20 mg/mL). The EC50 value calculated for antiradical activity ranged between 15.00 ± 1.06 µg/mL and 13.21 ± 3.24 µg/mL for ABTS and DPPH assays, respectively. Z. bungeana extracts were found to decrease the activity of tyrosinase by 50% (at 200 µg/mL) similarly to kojic acid and were slightly cytotoxic for human melanoma A375 cell line (at 200 µg/mL) with no effect on HaCaT keratinocytes. In the end, Z. bungeana did not reveal toxic effects in hemolytic assay as compared to the positive control Triton X-100. The performed tests show potential application of the plant in the treatment of infectious diseases, disorders caused by free radicals, and skin problems.

Pharmacological Evaluation of Artemisia cina Crude CO2 Subcritical Extract after the Removal of Santonin by Means of High Speed Countercurrent Chromatography.

Artemisia species are highly important due to their economic significance as medicines, fodder and food. Artemisia cina is an endemic species to Kazakhstan. In folk medicine, water extract of A. cina was used in the treatment of bronchial asthma while the alcohol extract has larvicidal and antituberculosis activity. The most common and most extensively studied compound from this species is the terpenoid santonin. The toxicity of this compound occurs at the doses of 60 mg for children and 200 mg for adults causing among other issues xanthopsia, leading to blindness. Having this in mind, the main idea of this work was to remove santonin from the crude extract and to check if the santonin-free extract would still be of any pharmacological importance. A CO2 subcritical extract was chromatographed using high-speed countercurrent chromatography (HSCCC) for the removal of santonin. The santonin-free CO2 subcritical extract (SFCO2E) as well as the isolated compound pectolinarigenin, a flavonoid, were assessed for their pharmacological actions. From the results obtained we can safely suggest that HSCCC is an efficient methodology to completely remove santonin from the CO2 subcritical extract. It was also possible to observe promising antinociceptive and anti-inflammatory activities for both SFCO2E and pectolinarigenin at concentrations that can justify the production of a phytomedicine with this endemic plant from Kazakhstan.

Quantification of santonin in eight species of Artemisia from Kazakhstan by means of HPLC-UV: Method development and validation.

Santonin, a powerful anthelmintic drug that was formely used to treat worms, is Artemisia cina's main constituent. However, due to its toxicity to humans, it is no longer in use. Kazakhstan is looking to introduce this plant as an anthelmintic drug for veterinary purposes, despite the known toxic properties of the santonin. The objective of this study was to develop a fast and specific method for the identification of santonin and its precise quantitation using HPLC-UV in order to avoid unnecessary intoxication, which is paramount for the development of veterinary medicines. The results obtained showed that santonin appears at around 5.7 minutes in this very reliable HPLC method. The validation of the method was performed by the investigation of parameters such as precision, accuracy, reproducibility and recovery. The method was used to identify and quantify santonin in leaves of A. scoparia, A. foetida, A. gmelinni, A. schrenkiana, A. frigida, A. sublesingiana, A terra-albae, and A. absinthium from Kazakhstan as well as in three different extracts of leaves of A. cina. This study has provided a faster and simpler method for the identification and quantification of this compound in other species of Artemisia of economic importance.