O cloridrato de raloxifeno é um fármaco utilizado para o tratamento e prevenção da osteoporose em mulheres na pós-menopausa e foi aprovado pelo FDA para prevenção de câncer de mama invasivo. Pertence à classe de moduladores seletivos de receptor estrogênico. Foi desenvolvido pela Ely Lilly Company e é comercializado como Evista® na forma de comprimidos na dosagem de 60mg. Devido à importância do fármaco e interesse do Instituto Nacional de Ciências e Tecnologia – Inovações Farmacêuticas no desenvolvimento métodos de síntese e de controle de qualidade, o presente trabalho teve como objetivo o desenvolvimento de métodos analíticos para assegurar a qualidade tanto da matéria-prima quanto do produto acabado. A matéria-prima foi analisada quanto às suas características físico-químicas através da determinação da solubilidade, do ponto de fusão e da calorimetria exploratória diferencial. A caracterização e identificação da matéria-prima foram realizadas identificando grupos característicos como o cloreto e grupo fenol e também por espectrofotometria na região do infravermelho e do ultravioleta e por cromatografia líquida de alta eficiência. A forma farmacêutica comprimidos também foi caracterizada e identificada através dos mesmos métodos que a matéria-prima e adicionalmente através de cromatografia em camada delgada e eletroforese capilar. Os métodos desenvolvidos e validados para quantificação da matéria-prima foram volumetria em meio não-aquoso e cromatografia líquida. Para os comprimidos, foram desenvolvidos métodos por espectrofotometria na região do ultravioleta, cromatografia líquida e eletroforese capilar. Os resultados de todos os métodos foram analisados estatisticamente para verificar equivalência nas determinações. / Raloxifene hydrochloride is used for the treatment and prevention of osteoporosis in post-menopausal women. It was approved by FDA for the prevention of invasive breast cancer. It was developed by Ely Lilly Company and marketed as Evista® in form of tablets of 60mg. Due to the importance of this substance and the interest of INCT-IF in its synthesis and quality control, the present work developed analytical methods to assure the quality of the raw substance and the pharmaceutical form. Qualitative and quantitative methods were developed for the analysis of raloxifene hydrochloride as raw substance and tablets. The raw substance was characterized by its physical and chemical characteristics by solubility, melting point and differencial scaning calorimetry. It was also identified by the analysis of characteristic groups such as chloride and phenol, and by infrared and ultraviolet spectrophotometry; also by high performance liquid chromatography. The tablets were also characterized and identified by the same methods as for the raw substance, but in addition by thin layer chromatography and capillary electrophoresis. The developed and validated methods for the assay of the raw substance were: non-aqueous titration and liquid chromatography. For the tablets, the assays were: ultraviolet spectrophotometry, liquid chromatography and capillary electrophoresis. The results of all methods were analyzed statistically to verify the equivalence of the determinations.

INTRODUÇÃO: Os polimorfismos do gene da proteína priônica (PRNP) podem estar associados a doenças neurológicas não priônicas. Estudos em pacientes com doença de Alzheimer (DA) apontam para possível associação entre os polimorfismos do códon 129 do PRNP e DA. Essa associação não foi estudada na população brasileira. Neste estudo, descrevemos a associação entre os diferentes polimorfismos do PRNP e DA. MÉTODOS: Foi estudada amostra composta por 100 pacientes com DA, acompanhados no Ambulatório de Neurologia Cognitiva e do Comportamento e no Centro de Referência em Distúrbios Cognitivos do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, pareados para grupo controle com 111 indivíduos, em relação à frequência dos diferentes polimorfismos do PRNP e o desempenho cognitivo. Os polimorfismos do PRNP foram estudados pelo método de cromotografia líquida de fase reversa desnaturante (DHPLC). Foi realizada extratificação da amostra pelo genótipo da apolipoproteína E (apoE). RESULTADOS: A frequência dos polimorfismos do códon 129 foi: 45,5% M/M, 42,4% M/V e 12,2% V/V nos pacientes com DA; e 39,6% M/M, 50,5% M/V e 9,9% V/V nos indivíduos controles (p=0,503). O códon 117 apresentou variante alélica silenciosa em 5% dos pacientes com DA e 3% dos controles (p=0,780). A deleção de um ocatapeptídeo repetido ocorreu em 5% dos pacientes com DA e 4% dos controles (p=0,738). Todos os pacientes com DA e os controles eram N171N. Uma paciente do grupo com DA apresentou a mutação V180I. A análise bivariada e regressão logística não mostraram associação entre os diferentes polimorfismos do códon 129 e o desempenho cognitivo nos pacientes com DA, assim como nos indivíduos cognitivamente normais. A extratificação segundo genótipo da apoE não revelou diferença em relação aos polimorfismos do códon 129 do PRNP entre os grupos DA e controles. CONCLUSÕES: Não houve diferença de frequência dos diferentes polimorfismos do códon 129 do PRNP entre os pacientes com DA e idosos cognitivamente normais, bem como em relação aos demais códons polimórficos do gene. Não houve diferença em relação ao desempenho cognitivo nos pacientes com DA e nos controles segundo o polimorfismo do códon 129 do PRNP. Um paciente apresentou mutação do códon 180 (V180I), e recebeu o diagnóstico de doença de Creutzfeldt-Jakob genética / INTRODUCTION: The polymorphism in the prion protein gene (PRNP) may influence non prion neurological diseases. Some reports associate Alzheimers disease (AD) and the polymorphic codon 129 of the PRNP. This association has not been studied in Brazilian population. In this study we aimed to describe the association between the polymorphisms of codon 129 of the PRNP and AD. METHODS: One hundred AD patients were evaluated in the Cognitive and Behavioral Neurology Unit and Cognitive Disorders Reference Center of the Hospital das Clínicas of the University of São Paulo School of Medicine, matched for 111 controls, regarding to the PRNP polymorphism and cognitive measures. The PRNP polymorphisms were analyzed using denaturing high-performance liquid chromatography (DHPLC). Analyzes stratifying by apoE genotype was performed. RESULTS: The distribution of the codon 129 polymorphisms were: 45.5% M/M, 42.4% M/V and 12.2% V/V in AD patients; 39.6% M/M, 50.5% M/V and 9.9% V/V in the control group (p=0.503). The 117 codon analysis revealed silent allelic variant in 5% of AD patients and 3% of controls (p=0.780). The octarepeat deletion occurred in 5% of AD and 4% of controls (p=0.738). All AD patients and controls were N171N. One AD patient had a point mutation at codon 180 (V180I). Logistic regression failed to confirm any association between AD cognitive performance and the codon 129 of PRNP, as well as in the control group. There was no association between the codon 129 genotypes and genotypes and AD according to the apoE stratification. CONCLUSIONS: There were no differences in the frequency of the codon 129 polymorphism between AD. control group, according to the codon 129 polymorphisms. A point mutation at the codon 180 (V180I) was diagnosed in one patient

<p>Statistical experimental designs have been used for method development and optimisation of separation. Two reversed phase HPLC methods were optimised. Parameters such as the pH, the amount of tetrabutylammonium (TBA; co-ion) and the gradient slope (acetonitrile) were investigated and optimised for separation of erythromycin A and eight related compounds. In the second method, a statistical experimental design was used, where the amounts of acetonitrile and octane sulphonate (OSA; counter ion) and the buffer concentration were studied, and generation of an α-plot with chromatogram simulations optimised the separation of six analytes.</p><p>The partial filling technique was used in capillary electrophoresis to introduce the chiral selector Cel7A. The effect of the pH, the ionic strength and the amount of acetonitrile on the separation and the peak shape of R- and S-propranolol were investigated.</p><p>Microemulsion electrokinetic chromatography (MEEKC) is a technique similar to micellar electrokinetic chromatography (MEKC), except that the microemulsion has a core of tiny droplets of oil inside the micelles. A large number of factors can be varied when using this technique. A screening design using the amounts of sodium dodecyl sulphate (SDS), Brij 35, 1-butanol and 2-propanol, the buffer concentration and the temperature as factors revealed that the amounts of SDS and 2-propanol were the most important factors for migration time and selectivity manipulation of eight different compounds varying in charge and hydrophobicity. SDS and 2-propanol in the MEEKC method were further investigated in a three-level full factorial design analysing 29 different compounds sorted into five different groups. Different optimisation strategies were evaluated such as generating response surface plots of the selectivity/resolution of the most critical pair of peaks, employing chromatographic functions, simplex optimisation in MODDE and 3D resolution maps in DryLab™.</p><p>Molecular descriptors were fitted in a PLS model to retention data from the three-level full factorial design of the MEEKC system. Two different test sets were used to study the predictive ability of the training set. It was concluded that 86 – 89% of the retention data could be predicted correctly for new molecules (80 – 120% of the experimental values) with different settings of SDS and 2-propanol.</p><p>Statistical experimental designs and chemometrics are valuable tools for the development and optimisation of analytical methods. The same chemometric strategies can be employed for all types of separation techniques.</p>

Within the field of depression the “monoamine hypothesis” has been the leading theory to explain the biological basis of depression. This theory proposes that the biological basis of depression is due to a deficiency in one or more of three key neurotransmitter systems, namely noradrenaline, dopamine and serotonin which are thought to mediate the therapeutic actions of virtually every known antidepressant agent. Citalopram is a selective serotonin-reuptake inhibitor (SSRI) used for the treatment of depression and anxiety disorders. Citalopram is a racemic compound, in other words composed of a 50:50 mixture of two enantiomers (S-(+)-citalopram and R-(-)-citalopram) and with one of the enantiomers (S-(+)-citalopram) accounting for the inhibitory effect. At the time of introduction of citalopram the physician needed a therapeutic drug monitoring service to identify patients with interactions, compliance problems and for handling questions concerning polymorphic enzymes and drug metabolism. An achiral analytical separation method based on solid-phase extraction followed by high-performance liquid chromatography (HPLC) was developed for routine therapeutic drug monitoring (TDM) of citalopram and its two main demethylated metabolites. As the data available on citalopram were from achiral concentration determinations and to be able to further investigate citalopram enantiomers effects and distribution, a chiral method for separation of the enantiomers of citalopram and its demethylated metabolites was established. The advances within chiral separation techniques have made measurement of the concentrations of the individual enantiomers in biological fluids possible. The process behind enantioselective separation is however not fully understood and the mechanism behind the separation can be further scrutinized by the use of multivariate methods. A study of the optimization and characterization of the separation of the enantiomers of citalopram, desmethylcitalopram and didesmethylcitalopram on an acetylated ß-cyclodextrin column, by use of two different chemometric programs - response surface modelling and sequential optimization was performed. Sequential optimization can be a quicker mean of optimizing a chromatographic separation; response surface modelling, in addition to enabling optimization of the chromatographic process, also serves as a tool for learning more about the separation mechanism. Studies of the antidepressant effect and pharmacokinetics of citalopram have been performed in adults, but the effects on children and adolescents have only been studied to a minor extent, despite the increasing use of citalopram in these age groups. A study was initiated to investigate adolescents treated for depression, with respect to the steady-state plasma concentrations of the enantiomers of citalopram and its demethylated metabolites. The ratios between the S- and R-enantiomers of citalopram and didesmethylcitalopram were in agreement with studies involving older patients. The concentrations of the S-(+)- and R-(-) enantiomers of citalopram and desmethylcitalopram were also in agreement with values from earlier studies. The results indicate that the use of oral contraceptives may have some influence on the metabolism of citalopram. This might be because of an interaction of the contraceptive hormones with the polymorphic CYP2C19 enzyme. Even though the SSRIs are considered less toxic compared with older monoamine-active drugs like the tricyclic/tetracyclic antidepressants, the risk of developing serious side effects such as ECG abnormalities and convulsions has been seen for citalopram, when larger doses have been ingested. Furthermore, fatal overdoses have been reported where citalopram alone was the cause of death. Data on the toxicity of each of the enantiomers in humans have not been reported and no data on blood levels of the enantiomers in cases of intoxication have been presented. An investigation was initiated on forensic autopsy cases where citalopram had been found at the routine screening and these cases were further analysed with enantioselective analysis to determine the blood concentrations of the enantiomers of citalopram and metabolites. Furthermore the genotyping regarding the polymorphic enzymes CYP2D6 and CYP2C19 were performed. In 53 autopsy cases, we found increasing S/R ratios with increasing concentrations of citalopram. We found also that high citalopram S/R ratio were associated with high parent drug to metabolite ratio and may be an indicator of recent intake. Only 3.8 % were found to be poor metabolizers regarding CYP2D6 and for CYP2C19 no poor metabolizer was found. Enantioselective analysis of citalopram and its metabolites can provide valuable information about the time that has elapsed between intake and death. Genotyping can be of help in specific cases but the possibility of pharmacokinetic interactions is apparently a far greater problem than genetic enzyme deficiency. / On the day of the public defence the status of article IV was: Submitted.

Statistical experimental designs have been used for method development and optimisation of separation. Two reversed phase HPLC methods were optimised. Parameters such as the pH, the amount of tetrabutylammonium (TBA; co-ion) and the gradient slope (acetonitrile) were investigated and optimised for separation of erythromycin A and eight related compounds. In the second method, a statistical experimental design was used, where the amounts of acetonitrile and octane sulphonate (OSA; counter ion) and the buffer concentration were studied, and generation of an α-plot with chromatogram simulations optimised the separation of six analytes. The partial filling technique was used in capillary electrophoresis to introduce the chiral selector Cel7A. The effect of the pH, the ionic strength and the amount of acetonitrile on the separation and the peak shape of R- and S-propranolol were investigated. Microemulsion electrokinetic chromatography (MEEKC) is a technique similar to micellar electrokinetic chromatography (MEKC), except that the microemulsion has a core of tiny droplets of oil inside the micelles. A large number of factors can be varied when using this technique. A screening design using the amounts of sodium dodecyl sulphate (SDS), Brij 35, 1-butanol and 2-propanol, the buffer concentration and the temperature as factors revealed that the amounts of SDS and 2-propanol were the most important factors for migration time and selectivity manipulation of eight different compounds varying in charge and hydrophobicity. SDS and 2-propanol in the MEEKC method were further investigated in a three-level full factorial design analysing 29 different compounds sorted into five different groups. Different optimisation strategies were evaluated such as generating response surface plots of the selectivity/resolution of the most critical pair of peaks, employing chromatographic functions, simplex optimisation in MODDE and 3D resolution maps in DryLab™. Molecular descriptors were fitted in a PLS model to retention data from the three-level full factorial design of the MEEKC system. Two different test sets were used to study the predictive ability of the training set. It was concluded that 86 – 89% of the retention data could be predicted correctly for new molecules (80 – 120% of the experimental values) with different settings of SDS and 2-propanol. Statistical experimental designs and chemometrics are valuable tools for the development and optimisation of analytical methods. The same chemometric strategies can be employed for all types of separation techniques.

The general aim of this thesis was to evaluate a newly designed and constructed miniaturized rotating disk apparatus for in vitro dissolution rate measurements of different drug substances from all of the classes in the Biopharmaceutical Classification System (BCS). The new equipment is based on a low volume flow-through cell of Plexiglas, a gold plated magnetic bar and a special designed press. The disk of drug substance (approx. 5 mg) is placed eccentrically in the bar. Rotation speeds were set with a graded magnetic stirrer. An external HPLC pump delivered a continuous flow of aqueous medium to the flow-through cell during dissolution testing. A reversed phase high-performance liquid chromatography system using diode array detection (RP-HPLC-DAD) was coupled online to the new equipment. The injections from the miniaturized rotating disk outlet into the quantifying HPLC system were controlled by a six-position switching valve. The injection volumes from the valve and the autosampler, used for the external standards, were statistically evaluated to match each other volumetrically. No analyses were longer than three minutes, using isocratic mode. A traditional USP rotating disk apparatus was used as a reference system and the two instruments were shown to be statistically dissimilar in the numerical dissolution rate values probably due to different hydrodynamics, but had approximately the same precision/repeatability. When correlating the logarithmic values of the in vitro dissolution rate (G) with the apparent solubility (S), using shake-flask methodology in the solubility studies, the two apparatuses gave the same correlation patterns. Further correlation studies were done where the media components were altered by the use of different buffer species or additives into the buffers, such as inorganic salts. Chemometric tools, e.g. orthogonal partial least squares (OPLS), were used to better evaluate the most influential factors for G and S in different media. The most significant factor for a model basic drug substance (terfenadine) was pH, followed by the ionic strength (I) and added sodium chloride in one of the media. However, the surfactants in the Fasted State Simulated Intestinal Fluid (FaSSIF-V2) were found to be insignificant for G and S in this study (using a 95% confidence interval). The new miniaturized apparatus is a promising prototype for in vitro dissolution rate measurements both for early screening purposes and in dissolution testing during drug development, but needs further instrumental improvements.

University of Macau / Institute of Chinese Medical Sciences

Meloksikamas, (4-hidroksi-2-metil-N-(5-metil-2-tiazolil)-2H-1,2-benzotiazin-3-karboksamid-1,1-dioksidas), yra nesteroidinis vaistas nuo uždegimo (NVNU), selektyvus ciklooksigenazės-II (COX-II) inhibitorius ir naudojamas reumatoidiniam artritui, osteoartritui ir kitoms sąnarių ligoms gydyti. Jis yra žymiai efektyvesnis ir sukelia mažiau šalutinių poveikių virškinamąjam traktui lyginant su kitais tradiciniais nesteroidiniais vaistais nuo uždegimo (NVNU). Gydomoji meloksikamo paros dozė (15 mg) yra viena mažiausių lyginant su kitais tradiciniais nesteroidiniais vaistais nuo uždegimo (NVNU). Europoje ir Lietuvoje yra registruotos tik peroralinės ir parenteralinės meloksikamo vaistų formos. Todėl siekiama sukurti transdermalines meloksikamo vaistų formas. Šio darbo tikslas – sukurti meloksikamo išsiskyrimo iš tirpalų pro dializės membraną in vitro metodą ir įvertinti išsiskyrusio meloksikamo kiekį. Tyrimo metu buvo naudojamas 1 proc. meloksikamo propileno glikolio tirpalas. Dializės membrana - regeneruotos celiuliozės. Akceptorinis tirpalas – buferinis tirpalas, kurio pH reikšmė 8,5. Palaikoma temperatūra - 37°C. Bandiniai buvo imami po 15, 30, 60, 120, 180 ir 300 minučių. Išsiskyręs meloksikamo kiekis iš bandinių kiekybiškai buvo vertinamas efektyviosios skysčių chromatografijos (HPLC) metodu. Didžiausias meloksikamo srautas (346, 963 µg/cm²/h) buvo po 15 minučių nuo eksperimento pradžios. Vėliau meloksikamo srautas mažėjo. Mažiausias meloksikamo srautas (67,401 µg/cm²/h)... [toliau žr. visą tekstą] / Meloxicam, (4-hydroxy-2-methyl-N-(5-methyl-2-thiazolyl)-2H-1, 2-benzothiazine-3-carbox-amide-1, 1-dioxide), a non-steroidal anti-inflammatory drug (NSAID) and selective cyclooxygenase-II (COX-II) inhibitor, is used in the treatment of rheumatoid arthritis, osteoartritis and other joint diseases. It has comparable efficiency and greater gastric tolerability in comparison to conventional non-steroidal anti-inflammatory drugs (NSAIDs). The efficiency dose of meloxicam (15 mg/day) is one of the lowest in the non-steroidal anti-inflammatory drugs (NSAIDs). There are registered only oral and parenteral formulations of meloxicam in Europe and Lithuania. So it is strive to create transdermal formulations of meloxicam. The aim of the present work was to develop in vitro meloxicam release through dialysis membrane method and measure the amount of released meloxicam. On release of meloxicam from 1 % meloxicam propylene glycol solution was used regenerated cellulose dialysis membrane. As acceptor fluid was used buffer pH 8,5. Temperature was controlled at 37°C. Samples 1 ml were withdrawn at intervals of 15, 30, 60, 120, 180 and 300 min. The quantitative determination of meloxicam in samples was performed by high performance liquid chromatography (HPLC). The maximum flux of meloxicam (346,963 µg/cm²/h) was after 15 min from the beginning of the experiment. Later on meloxicam flux was on the decrease. The minimum flux of meloxicam (67,401 µg/cm²/h) was after 300 min from the beginning of... [to full text]

Fenolinės rūgštys ir fenilpropanoidai pasižymi reikšmingu biologiniu aktyvumu ir yra vaistinių žaliavų veiklieji komponentai. Įteisintų tyrimo metodikų vystymas šių junginių nustatymui (kokybiniam ir kiekybiniam žaliavų tyrimui) yra ypač svarbus augalinių vaistinių preparatų efektyvumo, saugumo ir kokybės kontrolei. Iš skirtingų Lietuvos regionų buvo surinkta 18 propolio pavyzdžių. Atlikto darbo tikslas - analitiniais metodais ištirti Lietuvoje surinktų propolio pavyzdžių cheminę sudėtį, bei kiekybiškai ir kokybiškai įvertinti šias fenolines rūgštis ir fenilpropanoidus: chlorogeno rūgštį, cinamono rūgštį, galo rūgštį, ferulo rūgštį, kavos rūgštį, kumaro rūgštį, protokatecho (3,4-dihidroksibenzoinę) rūgštį, rozmarino rūgštį, vaniliną ir vanilino rūgštį. Atlikti tyrimai paprastos ir greitos atvirkštinių fazių efektyviosios skysčių chromatografijos metodikos optimizavimui ir įtiesinimui. Sukurto crhromatografinio metodo judrioji skystoji fazė yra metanolis ir 0,5 procento acto rūgšties tirpalas vandenyje. Esant optimalioms atskyrimo sąlygoms (aprašytoms skyriuje „Tyrimo metodas“), analizė atliekama 27 min., sulaikymo laikas 4,04 min (galo rūgštis), 6,93 min (protokatecho rūgštis), 11,30 min (chlorogeno rūgštis), 12,26 min (vanilinė rūgštis), 12,93 min (kavos rūgštis), 13,80 min (vanilinas), 16,97 min (kumaro rūgštis), 17,95 min (ferulo rūgštis), 22,29 min (rozmarino rūgštis) ir 26,30 min (cinamono rūgštis). Aptikimo riba ir nustatymo riba buvo atitinkamai tarp 0,004–0,021 ir 0,0... [toliau žr. visą tekstą] / The phenolic acids and phenylpropanoids have an important biological activity and are therapeutic agents of crude drugs. Development of validated analysis techniques of these phytotherapeutic agents (fingerprinting and assay procedures) is an important practice for efficacy, safety and quality control of herbal drug preparations. 18 samples of propolis were collected from different districts of Lithuania. The aim of the present work was to study chemical composition of collected samples and estimate analytical capabilities of the evaluation of selected phenolic acids and phenylpropanoids: caffeic acid, chlorogenic acid, cinnamic acid, coumaric acid, ferulic acid, gallic acid, protocatechuic (3,4-dihydroxybenzoic) acid, rosmarinic acid, vanillic acid and vanillin. Optimization and validation procedures of rapid and simple method of reversed phase high performance liquid chromatography were carried out. The mobile phase of the optimized chromatographic method consisted of methanol and 0.5 per cent acetic acid solvent in water. Under the optimal separation conditions (described in Methods), analysis was done in 27 min, with the retention times 4.04 min (gallic acid), 6.93 min (protocatechuic acid), 11.30 min (chlorogenic acid), 12.26 min (vanillic acid), 12.93 min (caffeic acid), 13.80 min (vanillin), 16.97 min (coumaric acid), 17.95 min (ferulic acid), 22.29 min (rosmarinic acid) and 26.30 min (cinnamic acid). The limits of detection and the limits of quantitation were between... [to full text]

Tyrimo tikslas - parinkti neetanolinio propolio ekstrakto technologiją ir atlikti veikliųjų medžiagų analizę. Uždaviniai: nustatyti propolio koncentracijos, ekstrakcijos tirpiklių bei jų koncentracijų, ekstrakcijos laiko, temperatūros, ekstrakcijos metodo įtakas fenolinių junginių išsiskyrimui. Nustatyti propolio ekstrakto antiradikalinį aktyvumą ir flavonoidų kiekį bei efektyviosios skysčių chromatografijos metodu nustatyti propolio ekstrakte esančius cheminius junginius ir jų antiradikalinį aktyvumą. Metodika. Bendras fenolinių junginių kiekis, buvo nustatytas naudojant farmakopėjinį Folino Ciocalteu metodą pagal galo rūgšties ekvivalentą. Flavonoidų kiekis nustatytas naudojant spektrofotometrinį metodą pagal rutino ekvivalentą. Antioksidacinis aktyvumas spektrofotometriškai buvo nustatomas naudojant DPPH radikalą. Efektyviosios skysčių chromatografijos su elektrocheminiu detektoriumi būdu buvo identifikuoti fenoliniai junginiai ir nustatytas jų antiradikalinis aktyvumas. Rezultatai. Didinant propolio koncentraciją, naudojant tirpiklius didėja bendras fenolinių junginių kiekis (p<0,05). Technologiniai procesai t.y. ekstrakcijos trukmė, temperatūros padidinimas daro reikšmingą įtaką fenolinių junginių išsiskyrimui (p<0,05). Didžiausias bendras fenolinių junginių kiekis 35,60 mg/ml buvo gautas naudojant ultragarsu skatinamą ekstrakciją 10 min esant 70 ⁰C temperatūrai, ekstrakcijos tirpikliu naudojant 30 proc. makrogolio ir vandens mišinį. Šiame ekstrakte nustatytas 88,76±1... [toliau žr. visą tekstą] / The aim of survey - to select the technology of nonethanolic propolis extract and analyse the active substances. Tasks: To examine the influence of concentration, solvent and its concentration, extraction time, temperature, the extraction method on release of total phenolic compounds. Determine the radical scavenging activity and flavonoid content of propolis extract and using high performance liquid chromatography method identify active substances and its antiradical activity. Methods: Total phenolic compounds were determined by using pharmacopoeial Folin Ciocalteu method expressed in gallic acid equivalents. Flavonoids were determined using a spectrophotometric method in rutin equivalent. Radical scavenging activity was determined by spectrophotometry using DPPH radical. High performance liquid chromatography with electrochemical detection were used to identify phenolic compounds and its radical scavenging activity. Results: Increasing of propolis concentration made a significance impact on the amount of total phenolic content (p<0,05). Extraction time, increase of temperature made a significance impact on increase of the amount of total phenolic content (p<0,05). The highest amount of total phenolic compounds 35,60 mg/ml were obtained using ultrasonic induced extraction for 10 minutes at 70 ⁰ C temperature and using 30 % macrogol – water solution as solvent. The radical scavenging capacity of this extract was 88,76 ± 1,27% and amount of flavonoids were 3,89 ± 0,43 mg/ml... [to full text]