[STUDY OF THE CYTOTOXIC ACTIVITY OF INDOLE ALKALOIDS FROM THE ABOVEGROUND ORGANS OF VINCA ROSEA L. INTRODUCED IN WESTERN GEORGIA].

The objects of this research were the aboveground and underground parts of Vinca rosea L., a species of periwinkle introduced in Western Georgia. The sum of alkaloids and biologically aactive fractions of monomeric alkaloids were obtained by liquid-liquid extraction, precipitation with petroleum ether (I), polybuffer distribution (II) and 10% acetic acid (III). 14 known compounds were identified by LC-MS/MS and GC/MS: Low molecular weight compounds (М+226, М+202, №1,2, М+168) and alkaloids Ajmalicine, Tetragidroalstonine, C20-dihydrova llesiahotamine, C19-C20 Vallesiahotaminole, Vallesiahotamine lacton, Polyneuridine, Pericy clivine, Lochnerame, Norharmane, Vidorosine, Vindolinine, Isovindolinine, Akuammicine. The cytotoxicity of monomeric alkaloids was evaluated for A-549 (cells of the lung cancer line), DLD-1 (cells of the rectal adenocarcinoma line) and W-1 (cells of the normal human fibroblast line). All three substances (I, II, III) showed pronounced cytotoxic activity.

STUDY OF THE BIOACTIVE COMPOUNDS CONTENT IN THE FLOWERS OF POLIANTHES TUBEROSA L. INTRODUCED BY GREEN TECHNOLOGIES.

The content of bioactive compounds of perennial, fragrant species of tuberose or Polianthes tuberosa L. representing the genus Polianthes L., asparagus (Asparagaceae Juss) family, highly decorative and widely accepted in the perfumery industry and extremely sensitive to environmental conditions, native to Mexico, was studied based on of raw materials of the flowers of the said plant introduced to five different locations of the BBG with various exposition and soil content by green technologies. Based on the analysis of the soil obtained from all five locations, we can conclude, that the growth and development of the plant along with the quality and amount of received raw materials significantly depend on the rate of soil acidity, humus, and the content of basic feeders. Good results were revealed in the conditions of the experimental plot of fragrant plants with well enough warmth, sunlight, and humidity. Applying 100% natural bio preparation GeoHumate had a positive influence on the plants on the experimental plot in terms of the growth and development of the plant and accumulation of bioactive compounds. After doing GC-MS researches of tuberose, Polianthes tuberosa L., flowers introduced by green technologies in five different locations of the BBG with various exposition and soil content, there were identified important compounds valuable for cosmetics, perfumery, medicine, and of different fields.

; INTRODUCTION OF AROMATIC PLANTS IN THE BATUMI BOTANICAL GARDEN AND THEIR RESEARCH FOR THE CONTENT OF BIOLOGICALLY ACTIVE COMPOUNDS.

The research objectives were to study the introduction process of different aromatic and spice plants in the Batumi Botanical Garden and the content of bioactive compounds in aromatic plants introduced and grown in the soil and climatic conditions of the garden.; Among the aromatic plants introduced to the BBG by the authors, the following species are having full vegetative and generative development in the open ground:Cuminum cyminum L.; Polianthes tuberosa L.; Iris pallida Lam.;Geranium macrorrhizum L., Piper suaveolens Ham., Piper piperita L., Thymus citriodorus Schreb., Satureja Montana L., Mentha piperita L., Origanum vulgare L., Mentha longifolia L., Hyssopus officinalis L.; Phyla scaberrima Moldenke, Ruta graveolens L.; Vegetative organs reach the full development, but without blooming:Curcuma longa L..; Zingiber officinale Roscoe.; Elettaria cardamomum Maton.; Coffea arabica L.; Coffea conephera L..; Vanilla planifolia Jacks.; Cassia acutifolia Delile.; For the purpose of studying the content of bioactive compounds in plants introduced to the soil and climatic conditions of the BBG with the help of green technologies, the following species were taken for analysis:Cassia acutifolia Delile. - seed, flower, leaf; Cuminum cyminum L. - seed; Coffea conefera L. andCoffea Arabica L., leaves and seeds; Vanilla planifolia Jacks. - leaves; Elletaria cardamomum Maton. -Cardamom leaves.With the help of gas chromatography - mass spectrometry GC-MS method, there were identified bioactive compounds of various classes. The content of essential oils is identified in all research objects.

CHARACTERIZATION OF SULPHIDE SILT PE-LOIDS IN SOME LAKES WITHIN ADJARA REGION AND DETERMINATION OF THE BIOLOGICALLY ACTIVE COMPOUNDS CONTENT.

Medicinal muds, so called Peloids in the last decade attract attention in many countries as effective means for the treatment- preventive, and perfumery-cosmetic purposes. It has been established that the possibilities of this formation of nature are not limited to the many-year experience of the mankind. In terms of further improvement of the human social sphere, nowadays peloids are referred to as XXI century material. The aim of the research was to study the physical-chemical characteristics, such as: humidity, pH, ash content, bulk (volumetric) density and organoleptic properties of sulphide silt peloids of some Adjara region lakes, as well as to determine the content of mineral and organic compounds (humic and organic acids). The objects of the study were the sulphide silt peloids of some Adjara region lakes: Mtsvane, Gasaliani, Shratiani, Nurigeli, Ardagani and Niphi. The main good quality indicators of the sulphide silt peloids have been determined according to the methods described in the State Pharmacopoeia XI. The mineral and organic compounds in the research objects have been determined using the modern instrumental methods of analysis described in the literature. Based on the performed studies, the main physical-chemical characteristics of sulphide silt peloids of some Adjara region lakes (Mtsvane, Gasaliani, Shratiani, Nurigeli, Ardagani and Niphi) have been studied and their mineral contents (Ca-Na Feldspar, K Feldspar, Ca- Montmorillonite, Hematite, Quartz, Magnetite, Chlorite) determined. By the spectrophotometric and gravimetric methods of analysis there is established, that the sulphide silt peloids of the lakes Ardagani (1,22%), Niphi (1,72%) and Gasaliani (1,33%) are distinguished by relatively higher content of humic acids. By the Gas Chromatography-Mass Spectrometry - method of analysis, in the study objects there is established the content of complex ethers of the following organic acids: Stearic, Palmitic, Caprylic, Myristic, Valeric, Lauric, Oxalic, Phthalic and Fumaric acids.

DEVELOPMENT OF THE G C -M S /M S METHOD FOR QUALITATIVE AND QUANTITATIVE DETERMINATION OF CLOZAPINE IN HUMAN BLOOD.

A simple, rapid, selective, sensitive gas chromatography-mass spectrometry (GC-MS/MS) method was developed and validated for the determination of clozapine in human blood. For isolation was used solid phase extraction. The calibration standards range was 5-1500ng/mL. Analysis was carried out on a gas chromatograph Agilent 7000A Quadrupole GC-MS/MS. GC run conditions - capillary column HP-5MS, 30m × 0.25mm, coated with a 0.25µm film, column temperature from 50°C to 305°C with an increase of 10°C/min; the injection port and the transfer line temperature were 310°C; carrier gas - helium at flow rate of 1.0 mL/min; Ionization was performed in the positive mode (EI+). Acquisition was made in TIC and MRM mode, electron impact 70Ev. Quantitative analysis was carried out by recording ions m/z 326 → 256, m/z 326 → 243 and m/z 326 → 192 for clozapine. The method was validated by investigating the following parameters: selectivity, linearity, accuracy, precision, identification of LOD and LLOQ and recovery. Method, given in this paper can be useful for determination of clozapine in forensic investigations and for TDM of schizophrenic patients to improve therapeutic efficacy during clozapine treatment.

DEVELOPMENT OF LC-MS/MS METHOD FOR DETERMINATION OF THE OLANZAPINE IN HUMAN PLASMA.

The study aim was to develop a validated selective, reproducible and sensitive Liquid Chromatography - Tandem Mass Spectrometry (LC-MS/MS) method for qualitative and quantitative determination of olanzapin (OLZ ) in human plasma. For isolation was used solid phase extraction with the Gilson GX-271 ASPEC Solid phase extraction system, cartridge Supelclean LC-18 SPE 500mg. Elution solvent contained 40mL of isopropanol, 4mL of concentrated ammonium hydroxide and 156mL of freshly prepared methylene chloride. Samples containing OLZ and IS were chromatographed on AGILENT TECHNOLOGIES 1290 Infinity AGILENT TECHNOLOGIES 6460 Triple quad LC-MS/MS system. Separation was performed by isocratic elution on Zorbax Eclipse plus C18 (100´4.5 mm, 3.8 µm) column, equipped with pre-column: UHPLC GUARD Zorbax Eclipse plus C18 (5×2.1 mm, 3.8 µm); column temperature was 35°C. The mobile phases consisted of 0.1 % water solution of formic acid HCOOH (H2O):0.1 % acetonitrile solution of formic acid HCOOH (CH3CN) in 70 :30 (v/v) proportion. The flow rate was 0.4 mL/min. Determination was performed on a triple-quadrupole mass spectrometer employing electrospray ionization technique (ESI+), operating in multiple reaction monitoring (MRM) and positive ion mode. Total chromatographic run time was 5 min. The optimized MRM transitions for OLZ were m/z 313 ® 256, m/z 313 →213 and m/z 313→198 as the quantitative and confirmative traces, respectively. Method was validated on following parameters: specificity, linearity, LLOQ, accuracy and precision, recovery, matrix effect and stability. The method was subsequently applied to in vitro plasma samples for determination of OLZ. The obtained data allows the opportunity for forensic investigation of the intoxication in postmortem cases.

Analysis of Perilla nankinensis decne essential oil using gas chromatography coupled with time-of-flight mass spectrometry.

Perilla is the genus of herbaceous plants of Lamiaceae family. The essential oil of Perilla is believed to be essential part of the herb. It was claimed to have anti-inflammatory, anti-hyperlipidemia, antioxidative and antimicrobial activities. Apart from pharmaceutical and nutrition purposes, It is an edible plant frequently used as a fresh vegetable. It was also applied to produce perfume and cosmetics. Due to the importance of the essential oil from the P. nanakinensis and the lack of detailed studies of them, our work aimed investigation of contents of essential compositions by GC- TOF/MS method. The materials of the study were the aerial parts of P. nankinesis collected at full flowering stage, Guria Region, Georgia. A qualitative analysis of the individual compounds in P. nankinensis essential oil performed by gas chromatography (GC) coupled with time-of-flight mass spectrometer (TOF/MS) for the identification of the resolved peaks. 28 components, which accounted for 77,7% of the oil, were identified. The main components of this essential oil were 1,3,6,10-Dodecatetraene, 3,7,11-trimethyl-, (Z,E)- (α-Farnesene) (34.3%), Caryophyllene oxide (10.2%), 1,6-Octadien-3-ol, 3,7-dimethyl- (linalool) (10.2%). Essential oil also contained humulene (3.9%),caryophyllene (1.9%), methyl salicilate (1.15%), 2-hexanoylfuran (1.0%), gernacrene (1.1%). Based on the results of our experimental data and literature reviews, Perilla essential oil can represent special interest and can be used as an important natural substance for obtaining medicinal and preventive products with the several biological activities.

Biological active compounds from Georgian Galanthus shaoricus.

Amaryllidaceae alkaloids exhibit antitumour, antiviral and anticholinergic activities. Some of them have been used in the treatment of myasthenia gravis, myopathy and diseases of the nervous system. In this study, the characterization of these compounds from Amaryllidaceae plants along with some biological activities and some regulations to conserve the native flora will be reviewed. Plants materials: Galanthus shaoricus Kem.-Nath., were collected in 2007-2008 during the flowering period in Georgia. The preparation of extracts and fractions were obtained using methanolic maceration. Crude alkaloidal extracts were typically purified by liquid-liquid partitioning of their basic forms in chloroform. Lycorine, galantamine and tazettine has been found as one of the major alkaloid from Amaryllidaceae plants. Galanthus shaoricus have shown good antimalarial and cytotoxic activity in a dose-dependent manner. Methanolic extracts from bulbs demonstrated significant growth inhibition on human Hela and HCT-116 cells lines with IC50 (µg/mL) 16.3±1.8; 22.1±2.9 (aerial parts) and 12.8±1.7; 16.5±1.9 (Bulbs), respectively. Concerning the Amaryllidaceae alkaloids, lycorine with IC50 (µM) 0.8±0.5 and 2.6±0.2, haemantaimene (IC50=1.1±0.7 and 2.7±0.8 µM), hamaine (IC50=3.4±1.0 and 6.2 ±1.4 µM), homolycorine (IC50=1.4±0.9 and 3.3±1.0 µM), hipeastrine (IC50=2.8±1.0 and 7.5±1.8 µM) were found to be responsible for the cytotoxic activity on HCT-116 and Hela cell lines, respectively.

Antioxidant polyphenols from Populus alba growing in Georgia.

The aim of the study was to evaluate natural antioxidant properties of Populus alba. The antioxidant effects in plants are mainly due to the presence of phenolic compounds such as flavonoids, phenolic acids, tannins and phenolic diterpenes. Oxidative damage is implicated in most diseases processes. In vitro studies are able to demonstrate for flavonols and flavones a considerable antioxidative activity, mainly based on scavenging of oxygen radicals. The antioxidative potential of different fractions (respective organic fractions of n-hexane, chloroform and ethyl acetate) of 70% methanol extract of Populus alba was evaluated using free radical-scavenging activity on DPPH (1,1-diphenyl-2-picrylhydrazyl ) assays. The methanol extract showed significant (p<0.05) activities in all antioxidant assays and contained a high level of total phenolic content. It was observed that the level of hydrophilic phenolic content was higher than that of hydrophobics. Among those organic solvent fractions, ethyl acetate fraction exhibited significant activities due to the highest level of total phenolic content and their IC50 values were 0.18±0.02 mg/mL, 0.26±0.03mg/mL and 0.28±0.02 mg/mL in DPPH, respectively. These activities were superior to those of a commercial natural antioxidants tested. The chloroform and hexan fractions also exhibited significant (p<0.05) free radical-scavenging activity, attributed to the high amount of hydrophilic phenolics.

Production of effervescent tablets Mucomali from polysaccharides of Malva Silvestris L.

The aim of the article was the estimation of efficiency of a method of production of effervescent tablets Mucomali from polysaccharides of Malva Silvestris L sum. The 5% polyethylene glycol solution of polyvinylpyrrolidone was used as connective agent. Experimentally was ascertained good solubility of effervescent tablets (Mucomali), produced from polysaccharide of Malva Silvestris L sum. The pressing power 110 mpa was found optimal for getting effervescent tablets of Mucomali.