International audienceTwo sesquiterpene lactones, grosheimin and cynaropicrin, present in the aerial part of the Chartolepis intermedia Boiss. plant can be obtained from the ethyl acetate solid extract of the dried plant. A large-scale purification processing 16 kg of dry plant is presented. The first liquid-liquid extraction step of the 16 kg produced 550 g of dry extract that were divided in two equal portions. A classical low-pressure chromatography method can produce from 275 g of dry extract 13.8 g of grosheimin and 52.3 g of cynaropicrin 95 % pure lactones in 110 h using large amounts of organic solvents and 8 kg of silica gel. Another purification method employing countercurrent chromatography, with a modern hydrostatic column, and reversed phase preparative liquid chromatography were used with the second 275 g dry extract portion. It could produce 17.9 g of 99.4 % pure grosheimin and 68 g of 98.7 % pure cynaropicrin lactones in 95 h, with better yields and minimized losses in chromatographic columns, and using smaller solvent volumes than the classical method. The two purification processes are fully described and compared

BACKGROUND: The medicinal plant of Thymus serpyllum L. in nature, depending on the geographical region, climatic conditions, and growing environment, is represented with some chemotypes. Composition and quantitative content of the basic groups of the biologically active substances can be differed, and thus their biological properties are also various. AIM: The aim of the study was to determine possibility of the using the ultrasonic extracts of two chemotypes of T. serpyllum L. of Central Kazakhstan as an antimicrobial agent against test strains of microorganisms. MATERIALS AND METHODS: Two samples of T. serpyllum were extracted with 70% ethanol using ultrasound. The polyphenol content of the ultrasound extracts was determined using the LC-ultraviolet-ESI- tandem mass spectrometry technique. A study of an antimicrobial activity of the ultrasonic extracts was performed with eight strains of Gram-positive bacteria, six strains of Gram-negative bacteria, and four cultures of fungi. RESULTS: The ultrasonic extracts of two chemotypes of T. serpyllum L. are similar in composition of phenolic compounds but differ in a quantitative content of phenolic acids and flavonoids, except for a rosmarinic acid. The ultrasonic extracts have a wide spectrum of antimicrobial activity, exhibit the bactericidal or bacteriostatic activity against all tested bacteria and fungi at a concentration of 0.0625–20 mg/ml, but differ in their strength of action against test strains of microorganisms. CONCLUSION: The ultrasonic extracts of two chemotypes of T. serpyllum L. of Central Kazakhstan can be considered as a potential drug with a wide spectrum of antimicrobial activity. The results of chromatographic analysis will be used for standardization of a drug.

molecular structure of which was confirmed by an XSA, were produced by reacting α-santonin and chlorine in aqueous acetonitrile. It was found that 2-chloro-α-santonin is highly effective against trichomonas.One of the most efficient synthetic pathways to chloro-derivatives of terpenoids is the reaction of terpene enones with gaseous chlorine [1][2][3]. Japanese researchers observed that the chemo-and stereoselectivity of the reaction of chlorine with α-santonin (1) depended on the nature of the solvent [1].In order to expand the list of available potentially biologically active chloro-derivatives of lactone 1, we carried out for the first time the chlorination of 1 in aqueous acetonitrile. Two reaction products, chloroepoxide 2 and chloride 3, were isolated in yields of 20 and 60%, respectively.The structures of chlorination products 2 and 3 were confirmed by UV, IR, and PMR spectra (see Experimental). The molecular structure of 3 was established by an x-ray structure analysis (XSA). It has the structure 2-chloro-3-oxo-7α,6,11β(H)-eudesm-1,4-dien-12,6-olide (Fig. 1).Six-membered rings A and B are pseudo-trans-fused. Ring A is planar within 0.02 Å. Ring B has the 7α,10β-chair conformation. Ring B is trans-fused to the lactone (C). Ring C has the 7α-envelope conformation. The conformation of 3 is similar to that of the previously studied 2-bromo-α-santonin [4]. Differences in the corresponding torsion angles are less than 2.8°. Table 1 gives the atomic coordinates of 3.The preparation in two steps of 3 from 1 by chlorination in CHCl 3 followed by dehydrochlorination in the presence of N,N-dimethylaniline has been described [1]. Four steps were required to prepare 2 [1].The reason that 3 forms upon chlorination in aqueous CH 3 CN is that the solvent itself is weakly basic. Scheme 1 shows the probable pathway for formation of 2. The appearance of the epoxy group in the 4,5-position can be explained by the presence of water in the reaction mixture. In order to check the correctness of this hypothesis, we carried out 10 model experiments using weighed portions (25 mg) of 1 for each experiment and CH 3 CN:H 2 O solvent mixtures with water content from 0 to 50%. Then, chlorination was carried out under identical conditions until starting 1 was fully converted. The solvent was distilled off, The solid was extracted with ethylacetate. The resulting extracts were left in vials until the solvent was completely evaporated. The solids were analyzed by HPLC. Table 2 gives the analytical results.

BACKGROUND: A possible reduction in stocks of medicinal plant raw materials of Thymus serpyllum L. and Thymus vulgaris L. leads to the need to expand the raw material base of the official medicinal plants with using of endemic species of the flora of Kazakhstan, in particular, Thymus rasitatus Klokov, and Thymus eremita Klokov. AIM: The aim of the study was to study the possibility of using 70% ethanol extracts of T. rasitatus and T. eremita as antimicrobial agents. MATERIALS AND METHODS: The aerial parts of T. rasitatus and T. eremita were extracted with 70% ethanol using ultrasound assisted extraction. The qualitative and quantitative analyses of the 70% ethanol extracts were determined using the liquid chromatography-detection-ESI-mass spectrometry-(MS)/MS technique. The study of the antimicrobial activity of these extracts was performed for eight strains of Gram-positive bacteria, six strains of Gram-negative bacteria, and four cultures of fungi. RESULTS: Chromatographic analysis of hydroalcoholic extracts of both investigated Thymus species showed very similar phenolic compounds composition. In both cases, the major components are luteolin-7-O-glucoside and rosmarinic acid. About 70% ethanol extracts of T. rasitatus and T. eremita have a broad spectrum of antimicrobial activity, exhibit the bactericidal or bacteriostatic activity against all tested bacteria and fungi at concentration range of 0.0195–20 mg/ml, but differ in their potency against tested strains of microorganisms. CONCLUSION: About 70% ethanol extracts of T. rasitatus and T. eremita, endemic plants in the flora of Kazakhstan, can be considered as potential drugs with a wide spectrum of antimicrobial activity. The results of chromatographic analysis could be used for drug standardization.

An HPLC method for quantitative determination of cynaropicrin in Centaurea scabiosa L. dry extract obtained using 40% aqueous ethanol has been developed and its performance has been validated. The proposed method is specific; ensures high accuracy, reproducibility, and linearity in an analytical range of ±30%; and can be used for reliable quality control of C. scabiosa L. dry extract and related preparations.