Zein instrumentazio eta material erosiko zenuke ingurugiroko analisiak egiten dituen laborategia muntatzeko?

Duración (en horas): Más de 50 horas. Destinatario: Estudiante y DocenteProiektu honen bitartez ikasleak prozedura analitikoan eman beharreko urratsak identifikatuko eta bakoitzak duen garrantzia ulertuko ditu. Urrats horien barnean sartzen dira laginketa eta laginaren aurretratamendua. Sarri prozedura analitikoa analisiarekin nahasten da eta ondorioz urrats honetan jartzen da esfortzu analitikorik handiena. Edonola ere, laginketa eta aurretratamenduaren urratsen prozedura analitiko osoaren errorerik altuenak (> %60) erakusten dituzte eta beharrezkoa da bi urrats hauen planifikazioak eta egokitasunak emaitzen kalitatera ekar dezakeen onura azpimarratzea. Proiektu honetan lagin solidoen eta lagin likidoen laginketarako tresneria, laginak gordetzeko berezitasunak, laginketarako estrategiak eta laginketari dagokion estatistika landu beharko du ikasleak. Aurretratamenduari dagokionez konposatu ez-organikoek zein organikoek (bide humela) erakusten duten teknika desberdinak aztertu beharko dituzte, hauen berezitasunak, aplikazioak, abantailak eta desabantailak justifikatuz. Eduki hauek lantzeko, ur-araztegi batek dituen hondakinak (urak eta lohiak) ingurugirora jaurtikitzeko araztegiak indarrean dagoen legedia betetzebetetzen duen ebaluatzeko prozedura analitikoaren bi urrats hauek definitu beharko ditu ikasleak

Chitosan-Coated Alginate Microcapsules of a Full-Spectrum Cannabis Extract: Characterization, Long-Term Stability and In Vitro Bioaccessibility

Cannabinoids present in Cannabis sativa are increasingly used in medicine due to their therapeutic potential. Moreover, the synergistic interaction between different cannabinoids and other plant constituents has led to the development of full-spectrum formulations for therapeutic treatments. In this work, the microencapsulation of a full-spectrum extract via vibration microencapsulation nozzle technique using chitosan-coated alginate is proposed to obtain an edible pharmaceutical-grade product. The suitability of microcapsules was assessed by their physicochemical characterization, long-term stability in three different storage conditions and in vitro gastrointestinal release. The synthetized microcapsules contained mainly ∆9-tetrahydrocannabinol (THC)-type and cannabinol (CBN)-type cannabinoids and had a mean size of 460 ± 260 µm and a mean sphericity of 0.5 ± 0.3. The stability assays revealed that capsules should be stored only at 4 °C in darkness to maintain their cannabinoid profile. In addition, based on the in vitro experiments, a fast intestinal release of cannabinoids ensures a medium–high bioaccessibility (57–77%) of therapeutically relevant compounds. The full characterization of microcapsules indicates that they could be used for the design of further full-spectrum cannabis oral formulations.This research was funded by the Basque Government through the financial support as consolidated group of the Basque Research System (IT1213-19 and IT1446-22)

Exploratory optimisation of a LC-HRMS based analytical method for untargeted metabolomic screening of Cannabis Sativa L. through Data Mining

Background Recent increase in public acceptance of cannabis as a natural medical alternative for certain neurological pathologies has led to its approval in different regions of the world. However, due to its previous illegal background, little research has been conducted around its biochemical insights. Therefore, in the current framework, metabolomics may be a suitable approach for deepening the knowledge around this plant species. Nevertheless, experimental methods in metabolomics must be carefully handled, as slight modifications can lead to metabolomic coverage loss. Hence, the main objective of this work was to optimise an analytical method for appropriate untargeted metabolomic screening of cannabis. Results We present an empirically optimised experimental procedure through which the broadest metabolomic coverage was obtained, in which extraction solvents for metabolite isolation, chromatographic columns for LC-qOrbitrap analysis and plant-representative biological tissues were compared. By exploratory means, it was determined that the solvent combination composed of CHCl3:H2O:CH3OH (2:1:1, v/v) provided the highest number of features from diverse chemical classes, as it was a two-phase extractant. In addition, a reverse phase 2.6 μm C18 100 Å (150 × 3 mm) chromatographic column was determined as the appropriate choice for adequate separation and further detection of the diverse metabolite classes. Apart from that, overall chromatographic peak quality provided by each column was observed and the need for batch correction methods through quality control (QC) samples was confirmed. At last, leaf and flower tissues resulted to provide complementary metabolic information of the plant, to the detriment of stem tissue, which resulted to be negligible. Significance It was concluded that the optimised experimental procedure could significantly ease the path for future research works related to cannabis metabolomics by LC-HRMS means, as the work was based on previous plant metabolomics literature. Furthermore, it is crucial to highlight that an optimal analytical method can vary depending on the main objective of the research, as changes in the experimental factors can lead to different outcomes, regardless of whether the results are better or worse.This work was financially supported by the Education Department of the Basque Country as a consolidated group of the Basque Research System (IT1213-19) and by Sovereign Fields S.L., in the framework of the project Metabolomic study of Cannabis Sativa L. cultivations and determination of contaminants in medical cannabis plants

Itsas ingurumenerako mikrokutsatzaile organiko hidrofobikoen presentzia eta eraginak aztertzeko estrategien taxutzea

The efects of hydrophobic microcontaminants such as alkylphenols, organophosphorus compounds, organochloride pesticides, phthalates and musk fragances were studied in mussels. The proposed analytical strategy is based on the combination of passive sampling studies, bioconcentration measurements of micropollutants in mussels, environmental NMR metabolomics of two tissues (gonad, muscle) and hemolymph of mussels and histological analysis. By means of this work, it was verified the feasibility of this strategy in controlled conditions with intent to use it in future environmental studies. The micropollutants mixture was responsible of early spawning in mussels. In fact, the most significant metabolic changes were observed in the same day that spawning took place.; Lan honetan, ingurumen analisirako metodologia planteamendu berri bat proposatu da. Horretarako, kimikariek eta biologoek baldintza kontrolatuetako esposizioa egiteko elkarlanean jardun genuen. Esperimentu honetan, muskuiluak eta lagin-biltze pasiboko tresnak hainbat mikrokutsatzaile hidrofoboren eraginpean jarri ziren. Batetik, kutsatzaileen kontzentrazioak neurtu ziren, lagin-biltze puntualen zein lagin-biltze pasiboen bitartez. Bestetik, muskuiluetan kutsatzaileen kontzentrazioak neurtu ziren. Biologoek, euren aldetik, muskuiluen azterketa histopatologikoa egin zuten. Azkenik, NMR-n oinarritutako metabolomika erabili zen muskuiluek kutsatzaileen aurrean ematen zuten erantzuna aztertzeko. Lau estrategien konbinazioarekin, kausa (kokteleko mikrokutsatzaileen kontzentrazioak eta muskuiluek metatutakoa) eta eragindako efektuak (histologia azterketetan behatutakoak) hobeto uler ditzakegu. Kutsatzaileek eragiten dituzten aldaketak ulertzeko eta ondorioak azaltzeko metabolomika baliabide erabilgarria dela ondorioztatu genuen. Kutsatzaileen koktelak errute goiztiarra eragin zien muskuiluei; izan ere, NMR metabolomika erabiliz, aldaketa metaboliko nabarienak errute-egunean ikusi ziren

Atzoko baliabideak, biharko osasuna: ardo-hondakinak eta kannabisa

Tesi hau bi zutabe nagusitan banatzen da. Lehenengoan, ardo-hondakinen balioa handitzeko metodologia bat garatu genuen. Lehenik, ardo-hondakinetatik polife-nolak eta gantz-azidoak, bakoitza bere aldetik, erauzi genituen jariakin gainkritikoak erabiliz. Polifenolak ez dira oso egonkorrak, ordea, eta gorputzean bioeskuragarrita-sun baxua dute. Arazo horiek konpontzeko, polifenolak mikrokapsularatzeko teknika bat garatu genuen. Bigarren zutabean, etorkizun hurbilean kannabisa sendagai bezala erabili ahal izateko, 3 urrats eman genituen. Alde batetik, landare ezberdinak efektu ez-berdinekin lotzea ahalbide dezakeen landareen kannabinoideen hatz-marka osatzeko analisi-teknika garatu genuen. Bestetik, gernuan eta plasman kannabinoideak eta euren metabolitoak kuantifikatzeko metodo bat ere garatu genuen. Azkenik, konposatu pu-ruen eta landare mota jakinen produkzioa optimizatzeko, kimiotipo ezberdineko landa-reen hazkuntzan zeharreko kannabinoideen eta terpenoen garapena aztertu genuen.; This thesis was divided in two lines of research. On the one hand, supercrit-ical fluid extraction was successfully applied to a sequential fractionation of fatty acids and polyphenols from wine wastes. Unfortunately, polyphenols have poor long-term stability and poor bioavailability, so they were microencapsulated. On the other hand, 3 different studies were done around medical use of cannabis. Firstly, in order to corre-late plants with their physiological effects, an analysis method was developed to estab-lish the cannabinoid fingerprinting of different cannabis plants. A method for the quan-tification of major cannabinoids and their metabolites in human urine and plasma was also developed. Finally, in order to optimize the production of bioactive compounds, the evolution of cannabinoid and terpene content of different chemotype plants during their growth was studied

Targeting the endocannabinoid system : future therapeutic strategies

The endocannabinoid system (ECS) is involved in many physiological regulation pathways in the human body, which makes this system the target of many drugs and therapies. In this review, we highlight the latest studies regarding the role of the ECS and the drugs that target it, with a particular focus on the basis for the discovery of new cannabinoid-based drugs. In addition, we propose some key steps, such as the creation of a cannabinoid–receptor interaction matrix (CRIM) and the use of metabolomics, toward the development of improved and more specific drugs for each relevant disease

Identification and quantification of cannabinoids in Cannabis sativa L. plants by high performance liquid chromatography-mass spectrometry

High performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) has been successfully applied to cannabis plant extracts in order to identify cannabinoid compounds after their quantitative isolation by means of supercritical fluid extraction (SFE). MS conditions were optimized by means of a central composite design (CCD) approach, and the analysis method was fully validated. Six major cannabinoids [tetrahydrocannabinolic acid (THCA), tetrahydrocannabinol (THC), cannabidiol (CBD), tetrahydrocannabivarin (THCV), cannabigerol (CBG), and cannabinol (CBN)] were quantified (RSD < 10%), and seven more cannabinoids were identified and verified by means of a liquid chromatograph coupled to a quadrupole-time-of-flight (Q-ToF) detector. Finally, based on the distribution of the analyzed cannabinoids in 30 Cannabis sativa L. plant varieties and the principal component analysis (PCA) of the resulting data, a clear difference was observed between outdoor and indoor grown plants, which was attributed to a higher concentration of THC, CBN, and CBD in outdoor grown plants. [Figure not available: see fulltext.

Kannabisa: ageriko altxor ezkutua

In this work 2 different studies have been done around medical use of cannabis. On the one hand, to help to correlate plants with their physiological effects, a HPLC-MS/MS method has been developed to establish the cannabinoid fingerprinting of different cannabis plants. On the other hand, in order to optimize the production of bioactive compounds, the evolution of cannabinoid and terpene content of different chemotype plants during their growth was studied. Concentration peaks for major cannabinoids and terpenes were establish and characteristic terpenes for each chemotype were identified.; Etorkizun hurbilean kannabisa sendagai bezala erabili ahal izateko bi urrats eman dira lan honetan. Alde batetik, landareak euren efektu fisiologikoekin lotzeko lagungarria izan daitekeen kannabinoideen hatz-marka ikertzeko metodo bat garatu da fragmentazio bikoitzeko masa-espektometriari akoplaturiko eraginkortasun handiko likido kromatografia bidez (HPLC-MS/MS High Performance Liquid chromatography-tandem mass spectrometry). Bestalde, kannabis landareko konposatuen produkzioa optimizatzeko kimiotipo ezberdineko landareen hazkuntzan zeharreko kannabinoideen eta terpenoen garapena aztertu da. Bertan, konposatu garrantzitsuenen kontzentrazio maximoak noiz agertzen diren ikusteaz gain, landare-mota bakoitzaren ezaugarri diren terpenoak zeintzuk diren aurkitu da

Evolution of the Cannabinoid and Terpene Content during the Growth of Cannabis sativa Plants from Different Chemotypes

The evolution of major cannabinoids and terpenes during the growth of Cannabis sativa plants was studied. In this work, seven different plants were selected: three each from chemotypes I and III and one from chemotype II. Fifty clones of each mother plant were grown indoors under controlled conditions. Every week, three plants from each variety were cut and dried, and the leaves and flowers were analyzed separately. Eight major cannabinoids were analyzed via HPLC-DAD, and 28 terpenes were quantified using GC-FID and verified via GC-MS. The chemotypes of the plants, as defined by the tetrahydrocannabinolic acid/cannabidiolic acid (THCA/CBDA) ratio, were clear from the beginning and stable during growth. The concentrations of the major cannabinoids and terpenes were determined, and different patterns were found among the chemotypes. In particular, the plants from chemotypes II and III needed more time to reach peak production of THCA, CBDA, and monoterpenes. Differences in the cannabigerolic acid development among the different chemotypes and between monoterpene and sesquiterpene evolution patterns were also observed. Plants of different chemotypes were clearly differentiated by their terpene content, and characteristic terpenes of each chemotype were identified