Anticoagulant activity of some Artemisia dracunculus leaf extracts.

Platelet hyperactivity and platelet interaction with endothelial cells contribute to the development and progression of many cardiovascular diseases such as atherosclerosis and thrombosis. The impact of platelet activity with different pharmacological agents, such as acetylsalicylic acid and coumarin derivatives, has been shown to be effective in the prevention of cardiovascular disease. Artemisia dracunculus, L. Asteraceae (Tarragon) is used for centuries in the daily diet in many Middle Eastern countries, and it is well known for its anticoagulant activity. The present study investigates the presence of coumarins in tarragon leaves and subsequently determines the extract with a major amount of coumarin derivatives. The solvents of different polarities and different pH values were used for the purpose of purifying the primary extract in order to obtain fractions with the highest coumarin content. Those extracts and fractions were investigated for their anticoagulant activity by determining prothrombin time (PT) and the international normalized ratio (INR), expressed in relation to the coagulation time of the healthy person. Purified extracts and fractions obtained from plant residue after essential oil distillation, concentrated in coumarin derivatives, showed the best anticoagulant activity, using samples of human blood. INR maximum value (2.34) and consequently the best anticoagulant activity showed the methanol extract at concentration of 5%.  The INR value of normal plasma in testing this extract was 1.05.

Identification and isolation of pharmacologically active triterpenes in Betuale cortex, Betula pendula Roth., Betulaceae.

Betulae cortex, Betula pendula Roth., Betulaceae, comprise triterpene substances which are confirmed to posses very important pharmacological activities such as anti-inflammatory, anticancer and antiviral. In this study, extraction of triterpene substances from both, inner and external birch bark was carried out and after that qualitative analysis on betulin, betulinic acid, oleanolic acid and lupeol was performed by method of thin layer chromatography. By this separation method, applying system for development benzene-ethyl acetate-formic acid (36:12:5), is gained a good separation of examined triterpene substances from methanol extracts of inner and external birch bark as well as used standards. From obtained row triterpene mixtures, certain triterpene substances are isolated using method of dry column chromatography. To those substances infrared (IR) spectra were recorded and compared with IR spectra of adequate standards. The study encloses all obtained IR spectra and interpretations on the basis of which can be concluded that triterpene substances, betulin, betulin acid and lupeol isolated from external birch bark give identical characteristic signals and absorbance as referent standards. Method of dry column chromatography has resulted as simple, efficient, repeatable and economical for laboratory conditions. Beside this, a sufficient quantity of examined triterpene substances is also obtained for continuation of their further analytical analysis.

Urinary hippuric acid after ingestion of edible fruits.

Aim of this study was to evaluate the biotransformation of simple phenols after ingestion of edible fruits and mixed food. It was analyzed hippuric acid in urine as biomarker of conjugation in the liver cells of glycine with aromatic phenolic acids such benzoic and salicylic acid from ingested food. Measurement of hippuric acid in urine samples of 10 healthy individuals: 5 female and 5 male with a mean age 51,5 years were recruited to participate in this study. Urine samples were collected for 24 hours. The additional meals 300 g of fruits: blueberry, cherry, raspberry, melon, blackberry and mixed food were given immediately before the 24 hr urine sampling. Otherwise, the meals given during 24 hr was a usually food. Biotransformation of phenols in edible fruits, that are together with liver glycins precursors of hippuric acid biosynthesis, was evaluated by direct spectrophotometric measurement of excreted hippuric acid in urine at 410 nm. It was established that the highest quantity of hippuric acid was after ingestion of 300 g of bilberry fruits (p< 0,003), and same quantity of cherries (p< 0,003). Concentration of excreted hippuric acid was twice higher after ingestion of these fruits in comparison with hippuric acid concentrations in urine after ingestion of common - mixed food. Quantity of biosynthesised hippuric acid was in direct correlation with the concentrations of its precursors, primarily phenol acids and other simple aromatic acids ingested with food.

Potential antimicrobial effects of pharmacognostic drugs.

Pharmacognostic aspect, in treating of natural medicinal materials and medicinal of natural origin, is presented by very complex group of analytical and applicative methods. They are applied by means of identification, purities and quality of natural medicinal materials of vegetal, animal or mineral origin, more precisely of pharmacognostic drugs. A great number of drugs and their isolated products show the antimicrobial activities. The chemical substances, which are present in drugs, are currier of antimicrobial activities manifested by action on different bacteria, fungus, parasites and even viruses. The whole series of methods for analyzing of antimicrobial effects of drugs and the medicines in general are treated by scientific discipline microbiology. In order to complete the pharmacognostic analyses of drugs and there medicinal products, the microbiological analyses take a very important role. The antimicrobial and antifungal effects of series of plant materials, of pharmacognostic drugs against the large number of bacteria and fungus Candida have been carried out. The drugs we analyzed were treated from their chemical point of view and that is the presence of: coumarins, mucus and anthraquinones. The contents of these substances in plant material were analyzed by method of thin layer chromatography that was published preliminary. The antimicrobial effects of analyzed drugs were determinate by using dilution method in antimicrobial analyses which have been carried out with decocts of drugs.

Thin layer chromatography-application in qualitative analysis on presence of coumarins and flavonoids in plant material.

Drugs, natural medicinal plant, animals and mineral materials, have a large and various application in official pharmacy and medicine. Carriers of multilateral pharmacological effects that those drugs shown, are chemically define as active components that are present in them. Methods of qualitative and quantitative analysis are used for the chemical investigation of components that drugs contain. Method of thin layer chromatography has been shown as very reliable. According to the chemical investigation of single drugs, it is possible to define a group of compound or single compound comparing them with standards. Relating to the usage of method of thin layer chromatography, it has been carried out investigation on presence of coumarins and flavonoids in domestic plant material that have wide everyday usage. Coumarins and flavonoids from the point of view of chemical belonging are phenol derivatives with important pharmacological effects. Applying method of thin layer chromatography, it is detected presence of coumarins and flavonoids substances in plant material that has been tested. Anethi graveolens fructus et folium (fruit and leaf of dill), Anethum graveolens L., Apiaceae, Avenae sativae fructus (fruit of oats), Avena sativa L., Poaceae and Asperulae odoratae herba (sweet woodruff), Asperula odorata L., Rubiaceae. Chromatograms are developed in systems cyclohexane-ethylacetat (13:7) and toluene-ether (1:1) saturated with 10% acetic acid, and visualisation by observing on UV lamp (254 and 366 nm), spraying with reagents KOH (10% ethanol solution) and diphenylboryloxyethylamine (1% methanol solution).