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

Abstract

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.

Keywords: Artemisia cina; HSCCC; anti-inflammatory; antinociceptive; pectolinarigenin; santonin-free CO2 subcritical extract.

Figure 1

Pectolinarigenin isolated from Artemisia cina CO₂ subcritical extract.

Figure 2

The results from the first phase of formalin of treated mice (santonin-free CO2 subcritical extract (SFCO₂E) at 10, 30, and 100 mg/kg or pectolinarigenin (Pecto) at 1, 3, and 10 mg/kg) were compared with mice that were given vehicle (blank), ASA (200 mg/kg) or morphine (2.5 mg/kg) used as positive controls. This figure shows the decrease of the bar against the untreated mice. The treatments which show a significant decrease in the bar size compared to blank are marked with (*), p < 0.05.

Figure 3

The results from the second phase of formalin of treated mice (SFCO₂E at 10, 30, and 100 mg/kg or pectolinarigenin at 1, 3, and 10 mg/kg) were compared with mice that were given vehicle (blank), ASA (200 mg/kg) and morphine (2.5 mg/kg) used as positive controls. This figure shows the decrease of the bar against the untreated mice. The treatments which show a significant decrease in the bar size compared to blank are marked with (*), p < 0.05.

Figure 4

The preliminary results of Pecto 1 (1 mg/kg of pectolinarigenin) using students t-test to analyse the data (* p < 0.05).

Figure 5

The pilot results of 1 (P) (1 mg/kg of pectolinarigenin) using students t-test to analyse the data. Treatment with Pectolinarigenin was not statistically significant.

Figure 6

The results from the hot plate model of treated mice (SFCO₂E at 10, 30, and 100 mg/kg) were compared with mice that were given vehicle (blank) and morphine (2.5 mg/kg) used as positive control. This figure shows the increase of the baseline against the untreated mice (blank). The treatments which show a significant increase in the baseline size compared to blank are marked with (*), p < 0.05. (#), p < 0.01.

Figure 7

The results from the total leukocytes migration to the air pouch of treated mice (SFCO₂E at 10, 30, and 100 mg/kg, pectolinarigenin at 1, 3, and 10 mg/kg or dexamethasone at 2.5 mg/kg were compared with mice that were given vehicle by oral gavage 60 min before carrageenan (1%, 1 mL) or PBS (1 mL) injection into subcutaneous sir pouch (SAP). This figure shows the decrease of the bar against the carrageenan injected group that received vehicle (p.o). The treatments that show a significant decrease in the bar size compared to the group receiving carrageenan are marked with (*), p < 0.05. (#), p < 0.01.