Inaug.-Diss.--Königsberg i. Pr. / At head of title: Aus dem botanischen und dem pharmazeutisch-chemischen Institut der Universität Königsberg. Vita. Includes bibliographical references.

Inaugural dissertation (Ph. D.)--Friedrich-Wilhelms-Universität zu Berlin, 1932. / Cover title. Bibliography : p.[53]

Czech Republic is one of the world's leading producers of the poppy seeds. Concerning the growing field area of poppy, the Czech Republic is lead producer of the poppy seeds at the international level (Vašák, 2010; Cihlář, 2012). Poppy seed is very sensitive on many internal and external factors during cultivation, especially in the beginning of germination and emergence. At present, there are not registered preparations available for seed dressing for poppy seed in the Czech Republic (Prokinová, 2006). However, there has been done many experiments based on testing of natural substances (eg. humates and plant extracts), microorganisms and especially the application of soil fungi in seed treatment recently The aim of this study is to evaluate the effect of selected regional varieties of poppy cultivation and seed treatment to yield parameters of poppy seed both individually and in combination,. For the thesis was essential to describe the suitability of varieties for growing in organic and integrated agriculture in relation to the efficiency of production. Evaluation of the seed quality of the varieties was carried out in the field as well as under laboratory conditions. Before sowing, seeds were treated by preparations Gliorex and TS Osivo, untreated variant has been used as a control. During plant cultivation were monitored shape of the plants, the degree of infestation, weed control and other treatments based on natural substances. Neem-Azal was used as the pest regulator. The second variant was treated by foliar application of TS Květa. Seeds harvested from variants of our trial were tested under laboratory conditions for comparison of differences between varieties. The tests also revealed the seed infestation and what percentage of the seed is viable. The last evaluation was carried out on the basis of the weight of thousands of seeds (HTS), which reflects the yield determined by the selected seed treatment.

The thesis deals with the study genetic diversity of poppies (Papaver somniferum L.) by SSR analysis. Altogether, 85 samples of poppies were evaluated. DNA was isolated from dried, mixed samples grown from seed using a modified CTAB method by Doyle and Doyle (1990). The seeds were provided by VÚO Opava. Six SSR primers were selected for the analysis and visualization was performed using chip electrophoresis. The PCR reaction was optimized with fluorescently labeled primer for fragmentation analysis. Chip electrophoresis was evaluated in MVSP using PCO analysis. The similarity matrix was further calculated and the samples showed hight genetic variability. The results of similarity matrix were provided to breeders as a basis for further breeding.

The reaction of tris(2,2’-bipyridyl)ruthenium(III) (Ru(bipy) ₃³⁺) with various analytes to generate chemiluminescence has been well documented. This investigation sought to undertake a chemiluminometic study of the reactions of Ru(bipy) ₃³⁺ with selected Papaver Somniferum alkaloids and specifically synthesised phenethylamines. The investigation, based on a kinetic study, primarily addressed the effect of varying reaction conditions (pH) on Ru(bipy) ₃³⁺ chemiluminescence production. To monitor these reactions, a batch chemiluminometer was specifically designed, fabricated and automated to conduct an extensive study on the selected compounds of interest. The instrumentation incorporated a custom built reaction cell and comprised an ‘on-line’ sample preparation system with which calibration standards could be automatically prepared. The instrumentation provided both time-independent (peak area) and time-dependent (kinetic profile) information. A novel approach to the stabilisation of Ru(bipy) ₃³⁺ as a chemiluminescencent reagent was also investigated and a recirculating system was employed with the batch chemiluminometer to provide a stable supply of Ru(bipy) ₃³⁺. Codeine, thebaine and 6-methoxy-codeine were the Papaver Somniferum alkaloids selected for this study and several N-methylated and N,N-dimethylated phenethylamines and methoxy-substituted phenetheylamines were also synthesised to investigate the affect of pH on the chemiluminescence emission efficiency. The versatility of the batch chemiluminometer facilitated the kinetic study of numerous analytes over a broad pH range. The exemplary performance of the chemiluminometer as an analytical instrument, was demonstrated by the calibration functions, based on peak area data, which exhibited excellent linearity and sensitivity. The estimated detection limits (3σ) for the selected alkaloids were in the range 2 x 10&<sup>-9<&/sup;> M to 7 x 10&<sup>-9<&/sup;> at pH 5.0 and above, which compared favourably to detection limits for the same compounds determined using FIA. Relative standard deviations (n=5) for peak areas ranged between 1% to 5% with a mean of 3.1% for all calibration standards above 2.5 x 10&<sup>-8<&/sup;> M. Correlation between concentration and peak area, irrespective of pH and analyte was excellent, with all but two calibration functions having r-squared values greater than 0.990. The analytical figures of merit exemplified the precision and robustness of the reagent delivery and ‘on-line’ sample preparation, as well as the sensitivity of the system. The employment of the chemiluminometer for the measurement of total chemiluminescence emission (peak area) was in itself a feasible analytical technique, which generated highly reproducible and consistent data. Excellent analytical figures of merit, based on peak area, were similarly achieved for the phenethylamines. The effects of analyte structure on chemiluminescence activity was also investigated for the alkaloids and the phenethylamines. Subtle structural variations between the three alkaloids resulted in either a moderately reduced or enhanced total emission that was two or three fold difference only. A significant difference in reaction kinetics was observed between thebaine and codeine/6-methoxy-codeine, which was dependent upon pH. The time-dependent data, namely the observed rate constants for the initial rise in intensity and for the subsequent decay rate, were obtained by fitting a mathematical function (based on the postulated reaction mechanism) to the raw data. The determination of these rate constants for chemiluminescence reactions highlighted the feasibility for utilising such measurements for quantitative analytical applications. The kinetic data were used to discriminate between analyte responses in order to determine the concentrations of individual analytes in a binary mixture. A preliminary, multi-component investigation performed on a binary mixture of codeine and 6-methoxy-codeine (1:1) successfully determined the concentrations of these individual components using such rate constant measurements. Consequently, variations in kinetics resulted in a significant difference between the relative chemiluminescence response based on peak area measurements and the relative response base on peak height measurements obtained using FIA. With regards to the observed reactivity of secondary amines and tertiary amines, chemiluminescence peak area determinations confirmed the vital role of pH on reaction efficiency, which was governed by structural features and kinetics. The tertiary amines investigated generally produced a greater emission under acidic conditions than the corresponding secondary amines. However, the measured chemiluminescence responses were highly dependent upon pH, with similar peak areas obtained for both amine groups under slightly alkaline conditions.

Diploma thesis with the topic: Formation of yield and dry matter with genetic resources of opium poppy (Papaver somniferum L.) was worked out at the Department of botanics and plant physiology, Faculty of agrobiology, food and natural resources ČZU in Prague. The purpose of this thesis was to determine genotype and ontogenetic differences in amount of yield, production of dry mass, and value of growth-analytic characteristics in certain species of opium poppy. Diploma thesis was based on the fact that most of the studies about opium poppy (Papaver somniferum L.) are focused on the content of alkaloids and other substances present in poppy seeds. Due to this facts, it is necessary to describe the genotypes of poppy plants and their genetic resources from the physiological point of view. Following species of opium poppy were chosen for the experiment: Akvarel, Albín, Buddha, Florian, Korneuburger Weisser, Lazur, Major, Marianne, Opál, Postomi, Sokol, Tatranský. These species were chosen according to their content of alkaloids, especially morfine very low content, low content, medium content and high content of morfine. All of the tested genotypes are tested as object of study of genetic poppy resources. Seeds were supplied by GB Oseva Pro s.r.o., o.z. VÚO Opava and company Český mák. Field experiments were established at the research laboratory FAPPZ ČZU in Prague, in Červený Újezd. This location is in Prague-West division, in area of Middle Bohemia. The altitude of this location is 401 metres above the sea level. The size of the field for testing was 2 m2. The experiment was based in 4 repetitions by the method of Latin square. Winter wheat was used as the preceding crop. Opium poppy was planted according to agrotechnical principles of planting. Content of dry mass in individual plant organs was measured in certain ontogenetic stages of opium poppy (35, 45, 49, 52, 54, 62, 81 BBCH). Size of leaf area was assessed by the image analysis WinDias. Yield of seeds was assessed as well. Growth-analytic characteristics were calculated from the values of wieght of dry mass and size of leaf area. The results confirmed that the size of plants is influenced by its ontogeny and also by its species. The results also confirm, that the height of plants is not influenced by the content of morfine. Connection between the colour of seeds and height of plant was not confirmed either. Increase in weight of above-ground biomass depending on ontogenetic stage and influence of plant species was confirmed. There are differences in production of yield in species of poppy with different contents of morfine because the average weight of yield in species with very low content of morfine was 112,36 g, in species with low content of morfine 110,41 g, in species with middle content of morfine 12167 g and in species with high content of morfine 123,16 g. It was approved that ontogeny of plants influences the leaf area index (LAI) as it increases until blooming and then it decreases as a result of senescence and falling off the old leaves. Differences in leaf area index were detected among the species of opium poppy, depending on the content of morfine. It was detected that crop growth rate (CGR), relative crop growth rate (Rw) and relative growth rate of assimilation area (RA) are influenced by the ontogeny of plants. It was detected that species with high content of morfine in poppy seeds (Buddha, Postomi, Lazur) and species with white seeds Sokol and Albín have lower yield than species with middle content of morfine.

Turkey is important major licid opium poppy (Papaver Somniferum) producer for medicinal and scientific purposes in the world and one of the two traditional producer country. The poppy seeds which are unique part of the opium poppy are used for food purpose. About 25.000 ton/year poppy seeds were produced for food purpose. Recent years morphine content of commercially available poppy seeds were speculated. Some studies have been reported that poppy seeds from different origins contain a wide variation of morphine (2 &ndash / 294) &mu / g/g content. They have been used also different sample preparation method (extraction and derivatization) and different instruments (GC-MS) during analyses. In this study morphine content was determined in Turkish origin which were white, yellow and blue poppy seeds colors by using high pressure liquid chromatography (HPLC). Different sample preparation method (without derivatization) was used. Morphine was eluted from poppy seeds by acidic water through stationary phase activated aluminum oxide. Poppy seed samples were collected from 13 provinces and 35 different sowing fields of Turkey. The determined morphine contents in poppy seeds were ranged between (9.73 to 37.46) &mu / g/g. Any study was not found in the literatures on determination of total phenolic in poppy seed. In this study, the optimum condition for extraction of polyphenols in poppy seeds was determined and then, total phenolic values were obtained in three different colours seeds by spectrophotometric method. At the end of experiments the total phenolic contents in white, yellow and blue colours poppy seed were found to be 4.44, 3.05 and 3.67 mg/g respectively.

Kaman, P.: Influence of plant biological protection on production of secondary metabolites of Papaver somniferum III. Diploma thesis. Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, Hradec Králové, 2018, 77 pp. Diploma thesis was aimed to verify the impact of biological protection on the production of alkaloids of the opium poppy (Papaver somniferum, L.). Polyversum biofungicide was selected for this purpose, the active ingredient of which is oomycete Pythium oligandrum. The poppy tested were Orbis and Lazur high-morphine varieties. Extracts were prepared from the individual samples which were analyzed by HPLC. The following alkaloids were morphine, codeine, papaverine, noscapine and thebain. The average morphine content was around 0,5 %. The largest amount of 0,62 % was found in the poppyhead of untreated variant of the Orbis variety. Codeine for both varieties averaged 0,07 % in the poppyhead of untreated variant. Treated plants reached slightly lower values. The average papaverine content was around 0,01 % in the treated and also untreated variant, but only in poppyhead of the Lazur variety. Higher noscapine content was measured in the Lazur poppyheads, averaging 0,06 %. The content was only around 0,01 % for Orbis variety. The alkaloid content was...

Bulvová, L.: Alkaloids of Papaver rhoeas L. (Papaveraceae) and their biological activity related to Alzheimerʼs disease I. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové 2017. The aim of this study was to process the summary alkaloidal extract of aerial parts of Papaver rhoeas L.; to isolate contained alkaloids using chromatographical methods; to identify them and to determine their inhibitory activity towards human enzymes acetylcholinesterase, butyrylcholinesterase and prolyloligopeptidase. Two alkaloids (+)-rhoeagenine and LB-2 were isolated, and the structure of LB-2 (its absolute configuration) is being determined nowadays. In vitro biological assays of these alkaloids found the following results: (+)-rhoeagenine (IC50 AChE ˃ 1000 μM, IC50 BuChE = 230 ± 10 μM, IC50 POP = 878 ± 45 μM) and LB-2 (IC50 AChE ˃ 1000 μM, IC50 BuChE = 314 ± 13 μM, IC50 POP = 706 ± 2 μM). The determined IC50 values of isolated alkaloids were compared with inhibitory standards of cholinesterases galanthamine (IC50 AChE = 1,71 ± 0,065 μM, IC50 BuChE = 42,30 ± 1,30 μM), huperzine A (IC50 AChE = 0,033 ± 0,001 μM, IC50 BuChE > 1000 μM, IC50 POP > 1000 μM) and rivastigmine (IC50 AChE = 0,037 ± 0,001 μM, IC50 BuChE = 0,0033 ± 0,0003...

Čakurdová, M.: Alkaloids of Papaver rhoeas L. (Papaveraceae) and their biological activity related to Alzheimer's disease II. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové, 2017. The aim of this work was focused on isolation of alkaloids from the fraction 1 (joined fractions 20-27), which was obtained from the summary alkaloid extract of Papaver rhoeas L. (Papaveraceae). The alkaloids were identified by structural analysis NMR, GC-MS, optical activity and melting point. Two alkaloids were identified as (-)-stylopine and (+)-rhoeadine. Isolated substances were tested on ability to inhibit the enzymes acetylcholinesterase, butyrylcholinesterase and prolyloligopeptidase. Obtained data were expressed as IC50 values: (-)-stylopine (IC50 AChE = 522 ± 67 µM, IC50 BuChE = >1000 µM, IC50 POP = >790 µM); (+)-rhoeadine (IC50 AChE = 915 ± 64 µM, IC50 BuChE = >1000 µM, IC50 POP = >790 µM). None of the isolated substances showed so potent cholinesterase inhibitory activity as the alkaloids galanthamine (IC50 AChE = 1.71± 0.065 µM, IC50 BuChE =42.30 ± 1.30 µM) and huperzine A (IC50 AChE = 0.033 ± 0.001 µM, IC50 BuChE = >1000 µM) or rivastigmine (IC50 AChE = 0.037 ± 0.001 µM, IC50 BuChE = 3.3 ± 0.3 nM)....