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Alkaloids of Dicranostigma franchetianum (Prain) Fedde and their selected biological activity
Wijaya, Viriyanata ; Opletal, Lubomír (advisor) ; Šmejkal, Karel (referee) ; Klouček, Pavel (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacognosy and Pharmaceutical Botany Candidate: Viriyanata Wijaya, M.Sc. Supervisor: Prof. RNDr. Lubomír Opletal, CSc. Title of Doctoral Thesis: Alkaloids of Dicranostigma franchetianum (Prain) Fedde and their selected biological activity Dicranostigma franchetianum (Prain) Fedde (Papaveraceae) is one of the representatives of the small genus Dicranostigma Hook. f. & Thomson. D. franchetianum (Prain) Fedde has been selected for the phytochemical investigation according to the screening study. In the primary screening of the alkaloid extract for cholinesterases inhibition, the inhibitory value was high (hAChE/hBChE, IC50 g/ml; 1.67 0.11 and 3.85 0.31) and together at least 15 alkaloids were found in the extract. The primary ethanol extract was prepared from 11.8 kg of dry herb (Garden of Medicinal Plants, Faculty of Pharmacy in Hradec Kralove). Using common chromatographic methods, 21 isoquinoline alkaloids of various structural types were isolated. All compounds have been identified using various spectrometric techniques (GC-MS, HPLC- MS, and 1D- and 2D-NMR, optical rotatory. The alkaloids obtained in sufficient amounts were determined for human acetylcholinesterase (hAChE), human butyrylcholinesterase (hBChE), and...
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Identification of aroma compounds from Cannabis
Jarošová, Šárka ; Klouček, Pavel (advisor) ; Polesný, Zbyněk (referee)
Hemp is a complex system of chemicals which interacts with one another. The most important chemicals in cannabis are cannabinoids and terpenoids. These compounds evince synergism that makes a number of medicinal effects, e.g. antioxidant, anti-inflammatory, antifungal and analgesic. Volatiles cause aroma of cannabis and have therapeutic effects when it is inhaled.
The main of this study was to identify sensory active compounds in the varieties of cannabis by GC-MS/FID and sensory analysis by GC-MS/O. There were 7 samples of varieties to analysis: Cannatonic (CT) Conspiracy Kush (CK) Jillybean (JB) Jacks Cleaner (JC), Jack Skellington (JS) Nordle (N) and Nurse Jackie (NJ). Selection of these varieties was based on its genetic origin - the ratio of morphological types C. sativa and C. indica. Aromatic compounds were extracted by using SPME fiber. Sensory analysis was performed by one trained sniffer.
The results show that the most represented volatiles in cannabis varieties (both in C. sativa and C. indica) is myrcene. This terpenoid was in the greatest amount in variety of N (33.4%), NJ (29.8%), CT (29.7%) and JB (29.7%). Then beta-caryophyllene had a large number of presence with high ratio in CK variety (21.1%), JC (20%) and NJ (14%). In the varieties with higher level of morphotype C. indica (CK, N), there were limonene and alpha-humulene represented. Results based on the sensory analysis showed, that sensory activity of limonene, linalool and alpha-humulene is higher in varieties with greater ratio of C. indica. The varieties with a higher ratio of morphotype C. sativa were rich in myrcene and terpinolene.
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Antimicrobial aktivity of cannabis extract in pathogens causing skin disease
Pařízek, Štěpán ; Fraňková, Adéla (advisor) ; Klouček, Pavel (referee)
Diseases caused by microbes are worldwide cause of morbidity and public health risks. Despite undisputed global benefits of antibiotics, the occurrence of infections caused by antibiotics-resistant bacteria becomes more frequent. This can be attributed to increased consumption of antibiotics in the long term, but also to their inappropriate use. Appart from that, progress in the field of synthetic antibacterial drugs research is dramatically stagnating in recent decades, plus the the antibiotics have bad effects to many bacteria types beneficial to human body. This generates a strong need to seek for natural remedies with similar or better antimicrobial effects. It is known that many plants produce a wide range of secondary metabolites with antimicrobial effects. Such metabilites inhibit growth and reproduction of pathogenic microorganisms by various mechanisms. From explored herbal medicines hemp, traditionally used to cure many diseases, appears to be a suitable alternative to antibiotics.
This thesis focuses on the antimicrobial activity of extracts of six different cannabis genotypes. Extracts with different ratios of cannabinoids have been tested using in vitro microdilution method on selected pathogens, causing skin disease (Staphylococcus spp., Streptococcus spp.).
Results confirmed the antimicrobial activity of all tested extracts. Very good minimum inhibitory concentration was assessed mainly for Streptococcus spp. with a concentration of 4 ug/ml. Extracts NHS4, JB3 and CK3 showed best results against all pathogens, with minimal inhibitory concentrations in the range from 4 to 16 ug/ml. Only extracts of CK2, N3 and JC1 have proven ineffective (MIC 512 ug/ml), because they could not be evaluated by the inhibitory concentration.
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Bioactive substance of Rheum race used in Traditional Asian Medicine.
Neradilová, Vanda ; Klusoňová, Hana (advisor) ; Klouček, Pavel (referee)
Vanda Neradiová Biologically active substances in the genus of Rheum species used in traditional Asian medicine Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Specialist in laboratory methods Background The goal of this study was to determine content substances and their quantity in two different species of genus Rheum and compare to what extent these two species differ in a number of content substances. Another criterion by which these species were compared was their place of origin. I have compared a species growing in its natural environment in the Himalayas with a second species growing in the Czech Republic. Methods For determination of content substances and their quantitative determination high - performance liquid chromatography with UV and fluorescent detection was used. Samples were compared with retention factors and retention times of standards. Furthermore, isolation and identification of probable physcion was carried out using mass spectrometry. Results By qualitative analysis of antraquinones, aloe - emodin, emodin, chrysophanol, physcion and rhein were determined and by analysis of stilbenes piceatannol and resveratrol were determined. Type R. acuminatum had greater representation of antharaquinons compared to R. australe. When comparing content...
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The Application of Essential oils Encapsulated into Nanoclay for Shelf-life Improvement of Fresh Chicken Meat
Vojtíšková, Zuzana ; Klouček, Pavel (advisor) ; Fraňková, Adéla (referee)
This dissertation aims to introduce essential oils as effective antimicrobial substances suitable for food preservation, focusing on the shelf life of meat. Meat is food which is - due to its high nutrient content - susceptible to microbial spoilage. Decaying of meat can be delayed by the selection of appropriate preservation methods. Nowadays there are increasingly used preservative modern methods which differ from classical preservatives procedures mainly in the consideration to the food. As very promising method can be considered preservation by essential oils. In the experimental part of this thesis the effect of encapsulated essential oils into the clay nanoparticles for the shelf life of fresh chicken meat is assessed. For the purpose of the experiment thyme essential oil and oil of savory in two different concentrations (oil:nanoclay - 1:5, 1:10 w/w) were used. Pieces of chicken meat (5 g) were inoculated with Salmonella enteritidis (ATCC 13076) and Escherichia coli (ATCC 25922) and then coated with the clay nanoparticles with the encapsulated essential oils. That treated meat was stored at 4 °C and assessed at 1st, 7th and 14th day of storage, together with three controls. Samples of the chicken meat were explored in terms of microbiological parameters and their inhibitive effect against pathogenic bacteria was examined. Samples were also subjected to sensory analysis, which should provide potential impact of essential oils on the organoleptic characteristics of meat. The results confirmed the antimicrobial activity of both essential oils against pathogenic bacteria. Thyme essential oil, however, showed greater inhibitory activity compared with oil of savory. Thyme essential oil has reduced in both tested bacteria their number in more than 3.5 log CFU.g-1 compared to the controls. Less favorable results were observed in the case of sensory analysis, because in terms of organoleptic characteristics the concentrations of oils were not accepted positively by the assessors. The samples treated with essential oils had undesirable bitter and pungent taste.
This work confirmed the antimicrobial activity of thyme and savory essential oils, which can be used for preservation of fresh meat. But the real application of the essential oils in food may be significantly limited by their impact on the organoleptic characteristics of the product.
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Laboratory evaluation of activity of traditional Bolivian medicinal plants
Hladová, Alena ; Klouček, Pavel (advisor) ; Přemysl, Přemysl (referee)
Traditional medicine of indigenous cultures is based on the medicinal effects of local plants. Those people have been taught to use these plants throughout history and they employ them as their primary health care. This is supported by the fact that the natives do not have the financial resources to pay for medical care and at the same time it is also accompanied by poor availability of hospitals in areas where these cultures are located.
This thesis covers laboratory verification of antimicrobial and antioxidant activity
of Amazonian plants from south west Bolivia. This paper also summarizes the economic and political situation of the country together with the development of traditional medicine in the world and with the potential use of this medicine in pharmacy. Part of the work is devoted
to the issue of ever-increasing antimicrobial resistance and the possible effects of free radicals in the human body. The experiment was performed by testing nine samples of imported dried plants in laboratory. To determine the antimicrobial properties a microdilution method was used and the antioxidant activity was verified with the ORAC method which is based
on the elimination of free radicals. The microdilution method confirmed 80% of the MIC
in the initial concentrations tested (1,024 mg/ml) in Satureja ovata plants against Staphylococcus aureus ATCC 33591 bacteria and Streptococcus pyogenes bacteria and in the plant Baccharis genistelloides against S. pyrogenes bacteria. The other samples have failed
to provide the value of the minimum inhibitory concentrations. This can be attributed
to several factors which are described in this thesis. In contrast the positive effect free radicals was confirmed in all samples. The highest efficacy showed Satureja ovata (1373,2 ug TE/mg extract). Writing this thesis was accompanied by a lack of information about the tested plants, both in terms of their botanical description and in terms of comparing the laboratory results obtained with the results of specialized studies. Therefore the results of the experiment were studied in comparison to traditional use of the tested plants that have been provided
by Bolivian natives or their effects have been analyzed from the literature.
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Laboratory evaluation of activity of traditional Ecuadorian medicinal plants
Klepáčková, Kateřina ; Klouček, Pavel (advisor) ; Přemysl, Přemysl (referee)
Health care in Ecuador varies greatly depending on socioeconomic status, age, sex and place of residence. One third of the population has no regular access to health services and more than two-thirds of the population has no health insurance. Herbal medicine is
the synthesis of therapeutic experience of indigenous physicians which has been evolving
for hundreds of years and its resources are used to prevent and treat various types of diseases. Plants are popular natural remedy due to their efficacy, safety and minimal side effects. However, certain combinations of herbal medicine and drugs can be dangerous. Sciences, which help to examine the context of the traditional use of plants in the timeframe and also contribute to the discovery of their bioactive compounds, are called ethnobotany and ethnopharmacology. This paper investigates ten plants imported from Ecuador, which are traditionally used for healing various types of diseases such as gastrointestinal disorders, influenza, cold, urinary tract infection, skin diseases in the pelvic cavity etc. Efficiency research of herbal medicine has a great significance for future healthcare because
of the expansion of antimicrobial resistance, which is currently becoming a serious health risk for both humans and animals. Antimicrobial and antioxidant activity of plants, which are
in this thesis measured by the broth microdilution method and ORAC method, are important indicators for the analysis of medicinal plants. The only demonstrable positive result
of testing the antimicrobial activity was the discovery of minimum inhibitory concentration
of Espeletia schultzii against Streptococcus pyogenes (MIC=0.256 mg/mL). The highest antioxidant activity was discovered in the plant Croton elegans Kunth (743,64 +- 28,56 microgram of trolox equal to 1 miligram of the extract), for which no records associated to antioxidant activity were found.
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Selection of cannabis chemotypes suitable for therapeutic use with predominant active substance THC
Michnová, Lenka ; Fraňková, Adéla (advisor) ; Klouček, Pavel (referee)
During recent years the possible medical application of cannabis has been discussed more and more. It turns out, that the active substances (cannabinoids cannabidiol and tetrahydrocannabinol) have a significant potential in medicine and do not have to be only misused as a recreational drug. The Department of Crop Production (Faculty of Agrobiology, Food and Natural Resources) is the first institution with granted permission to legally cultivate cannabis, with higher THC content than 0,3 %, for research purposes.
The first part of literate review recapitulates the basic knowledge about the cannabis, its biologically active substances and current legislation. The second part describes the endocannabinoid system and utilization of cannabis in the field of medicine. This thesis aims to evaluate the THC and CBD content in seven different cannabis cultivar strains (Nurse Jackie, Conspiracy Kush, Jillybean, Jack Cleaner 2, Jack Skellington, Nordle and National Health Service). The cannabis plants were cultivated in three cultivation cycles. The content of active substances in dried and homogenised inflorescence was analysed by gas chromatography with flame ionization detection (GC-FID) according to UN methodology. Based on the analysis, the appropriate genotypes were chosen, which are in accordance of Regulation 236/2015 Sb. and needs of the patients.
According to results it was found, that all seven analysed cultivars are suitable for medical utilization regarding the content of THC and CBD. The highest average content of THC was measured by cultivar Jack Cleaner 2. On the other hand, the lowest THC content was established by cultivar Nordle. It was demonstrated, that the differences in THC content are connected rather to genotype variation than cultivation factors. The main goal of the thesis was to discover which of the seven cultivars will be stable regarding not only THC and CBD content, but also the crop yield. In those regards the research shows the cultivar Jack Cleaner 2 and Nurse Jackie as the most suitable. Preliminary results indicate that these two cultivar strains would be the most suitable for the therapeutic purposes.
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Influence of Essential Oil Blanching on Drying Kinetics of Dehydrated Meat
Claramount Ruiz, Diego Antonio ; Banout, Jan (advisor) ; Klouček, Pavel (referee)
Meat is one of the most valuable livestock products, for many people serves as their first source of animal protein. Rising demand for meat in developing countries combined with poor access to synthetic antioxidants necessary for proper food preservation along with their potential side-effects highlights the importance of feasible alternatives for rural populations.
Among the most critical components that cause quality deterioration are the lipids and proteins oxidation, in time if not treated properly this will cause off-odor and off-flavor production in the meat, for that reason this dissertation focus on the application of Essential Oil (EO) as alternative to those more commonly used synthetic antioxidants like butyl hydroxyanisole (BHA) and butylhydroxytoluene (BHT) as pre-treatment of dehydrated meat, and its further effects if any during the drying process.
Beef meat samples will be prepared, and tree different types of pretreatment will be studied along with a combination of three possibilities as concentration of Essential Oil, the three types of pre-treatment will be ; Steam Blanching (SB), Hot Air Blanching (HAB) and Oil Treatment (OT), this were combined with different concentrations of Oregano EO, those were 1.5ml, 3ml, and 6ml. The approach will focus on finding if there was any correlation between the concentration of Oregano EO used versus the speed of drying and if there is any correlation between the different types of pretreatment applied.
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Quality of potato chips made of potatoes treated with carvone
Gildainová, Dana ; Klouček, Pavel (advisor) ; Fraňková, Adéla (referee)
This diploma thesis evaluates the ability of a monoterpene S-carvone (main component of caraway oil) to inhibit the sprouting of stored potatoes intended for further processing into a food products. Then it evaluates the influence of carvone's special treatment on selected qualitative parameters of potato chips.
For this experiment, two concentrations of carvone were selected (8 and 32 microliters per liter), applied altogether 4-times on stored potatoes within 3-week intervals. For this application a modified chromatograph was used to ensure a uniform dispersion of the compound. Special treatment was being done for a period of 10 minutes, while each variant of this treatment had 3 repetitions. After each treatment the inhibition effect of carvone on the growth of sprouts was evaluated. Also, there was an evaluation of the content of reducing sugars using the Luff-Schoorl method. Potato chips were repeatedly produced from the treated potatoes. Instrumental measuring of the texture of these potato chips was done on a testing device Instron. Potato chips were also a subject to a sensory evaluation to find out the degree of influence on organoleptic properties.
On the basis of the results, the inhibition effect had been confirmed only in the treatment by carvone with a concentration 32 microliters per liter. The variant of treatment by 8 microliters per liter provided the same results as the control variant. Content of reducing sugars in a tuber had not been influenced by a treatment by S-carvone. During the instrumental measuring of textural properties, a significant difference between control and variants treatment by carvone had been confirmed only after the second and fourth treatment. These chips had shown higher maximal strength needed for crunching of potato chip in comparison with the control. On the basis of the sensory profile of potato chips, no organoleptic properties had been influenced even after the fourth treatment with carvone of stored potatoes. Essential oils and its active substances represent a suitable substitution for used synthetic substances, because there had been recognized no negative effect of carvone on the qualitative parameters.
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