Extraction Comparison of Cannabis sativa inflorescences
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Orbitrap Exploris 120, 5975 Agilent GC-MS
ORGANISM(S): Cannabis Sativa (ncbitaxon:3483)
SUBMITTER: 
Josh J Kellogg
PROVIDER: MSV000094650 | MassIVE | Wed May 01 04:34:00 BST 2024
REPOSITORIES: MassIVE
Similar Datasets
Project description:Isolation of the therapeutic cannabinoid compounds from Cannabis Sativa L. (C. Sativa) is important for the development of cannabis-based pharmaceuticals for cancer treatment, among other ailments. The main pharmacological cannabinoids are THC and CBD. However, THC also induces undesirable psychoactive effects. The decarboxylation process converts the naturally occurring acidic forms of cannabinoids, such as cannabidiolic acid (CBDA) and tetrahydrocannabinolic acid (THCA), to their more active neutral forms, known as cannabidiol (CBD) and tetrahydrocannabinol (THC). The purpose of this study was to selectively extract cannabinoids using a novel in situ decarboxylation pressurized hot water extraction (PHWE) system. The decarboxylation step was evaluated at different temperature (80-150 °C) and time (5-60 min) settings to obtain the optimal conditions for the decarboxylation-PHWE system using response surface methodology (RSM). The system was optimized to produce cannabis extracts with high CBD content, while suppressing the THC and CBN content. The identification and quantification of cannabinoid compounds were determined using UHPLC-MS/MS with external calibration. As a result, the RSM has shown good predictive capability with a p-value < 0.05, and the chosen parameters revealed to have a significant effect on the CBD, CBN and THC content. The optimal decarboxylation conditions for an extract richer in CBD than THC were set at 149.9 °C and 42 min as decarboxylation temperature and decarboxylation time, respectively. The extraction recoveries ranged between 96.56 and 103.42%, 95.22 and 99.95%, 99.62 and 99.81% for CBD, CBN and THC, respectively.
| S-EPMC8199533 | biostudies-literature
Project description:Even if a large amount of high-throughput functional genomic data exists, most researchers feature a strong background in molecular biology but lack advanced bioinformatics skills. In this work, publicly available gene expression datasets have been analyzed giving rise to a total of 40,224 gene expression profiles within different Cannabis tissues/developmental stages. The resource here proposed will provide researchers with a starting point for future investigations of Cannabis sativa.
2017-01-13 | GSE93201 | GEO
Project description:Background Ribosome depleted RNA-seq is well suited for understanding coding- and noncoding transcript expression in clinical samples with diverse input RNA quality and mass. Current choices for RNA extraction include phase-separation (e.g., ThermoFisher TRIzol) approaches to column-based (e.g., Norgen Spin Columns) approaches. Each method, however, has been noted to have differing effects on the properties of the extracted nucleic acids, precluding the straightforward bioinformatic comparison of data generated by the two methods. Methods In this study, we present differences in data differing only in total RNA extraction approaches (phase separation vs. column) from clinical samples processed by the same tissue bank and sequencing facility personnel. We use data from seven samples with libraries generated from RNA extracted with both extraction methods to characterize the effect of ‘differentially extracted’ genes (DExGs) on transcriptomic profiles. We then use another set of 148 samples extracted with one or the other method to confirm the differences observed in the 7-sample dataset. Results The large number of samples employed in this study allows us to find significant differences in important quality control parameters as well as many DExGs between the two methods. We identify several biophysical properties which are potential drivers of these differences. Conclusions We show that partial correction of extraction method mediated differences can be achieved but full correction is not possible. These results lay the foundation for principled decision making of RNA extraction methods for use in clinical samples in a translational environment, particularly in light of recent reagent shortages during the COVID-19 pandemic.
2023-09-28 | GSE158705 | GEO
Project description:BackgroundAllergic sensitization to Cannabis sativa is rarely reported, but the increasing consumption of marijuana has resulted in an increase in the number of individuals who become sensitized. To date, little is known about the causal allergens associated with C sativa.ObjectiveTo characterize marijuana allergens in different components of the C sativa plant using serum IgE from marijuana sensitized patients.MethodsSerum samples from 23 patients with a positive skin prick test result to a crude C sativa extract were evaluated. IgE reactivity was variable between patients and C sativa extracts. IgE reactivity to C sativa proteins in Western blots was heterogeneous and ranged from 10 to 70 kDa. Putative allergens derived from 2-dimensional gels were identified.ResultsProminent IgE reactive bands included a 23-kDa oxygen-evolving enhancer protein 2 and a 50-kDa protein identified to be the photosynthetic enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase. Additional proteins were identified in the proteomic analysis, including those from adenosine triphosphate synthase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and luminal binding protein (heat shock protein 70), suggesting these proteins are potential allergens. Deglycosylation studies helped refine protein allergen identification and demonstrated significant IgE antibodies against plant oligosaccharides that could help explain cross-reactivity.ConclusionIdentification and characterization of allergens from C sativa may be helpful in further understanding allergic sensitization to this plant species.
| S-EPMC3726218 | biostudies-literature
Project description:Cannabis (Cannabis sativa) plants produce and accumulate a terpene-rich resin in glandular trichomes, which are abundant on the surface of the female inflorescence. Bouquets of different monoterpenes and sesquiterpenes are important components of cannabis resin as they define some of the unique organoleptic properties and may also influence medicinal qualities of different cannabis strains and varieties. Transcriptome analysis of trichomes of the cannabis hemp variety 'Finola' revealed sequences of all stages of terpene biosynthesis. Nine cannabis terpene synthases (CsTPS) were identified in subfamilies TPS-a and TPS-b. Functional characterization identified mono- and sesqui-TPS, whose products collectively comprise most of the terpenes of 'Finola' resin, including major compounds such as β-myrcene, (E)-β-ocimene, (-)-limonene, (+)-α-pinene, β-caryophyllene, and α-humulene. Transcripts associated with terpene biosynthesis are highly expressed in trichomes compared to non-resin producing tissues. Knowledge of the CsTPS gene family may offer opportunities for selection and improvement of terpene profiles of interest in different cannabis strains and varieties.
| S-EPMC5371325 | biostudies-literature
Project description:A comparison of sequencing platforms and approaches for arbovirus sequencing
| PRJEB47177 | ENA
Project description:Serum miRNAs are considered useful as non-invasive biomarkers for various diseases, but the optimal method for extracting RNA from serum is currently unknown. In this study, several RNA extraction kits were used to determine which kit is the optimal method. RNA was extracted from the serum of 8-week-old C57BL/6NJcl male mice according to the protocol of each RNA extraction kit. The yield of extracted RNA samples was calculated and electrophoretic patterns were evaluated by Agilent bioanalyzer. Expression patterns of the extracted RNA samples were confirmed by Agilent mouse miRNA microarray. The results showed significant differences in RNA yields in the miRNeasy serum/plasma advanced kit, and mirVana™ PARIS™ RNA and Native Protein Purification Kit compared to almost all other samples. Furthermore, two peaks were identified in the miRNeasy serum/plasma advanced kit using small RNA kit of Agilent bioanalyzer, one at 20-40 nucleotides (nt) and the other around 40-100 nt whereas the other reagents had a single peak. In addition, a high correlation was observed between the two RNA extraction kits in microarray. These results suggest that the above two kits are suitable for miRNA extraction from mouse serum.
2024-01-01 | GSE246437 | GEO
Project description:The goal of the present project was to investigate the applicability of a triple extraction (”TE”), which yields RNA, DNA and protein in a single procedure, for gel-free mass spectrometry-based proteomics. Proven successful for large-scale transcriptomics and genomics, we examined TE compatibility with mass spectrometry-based proteomics and phospho-proteomics by comparison to a urea standard processing. For whole proteome, peptides were fractionated using SAX; for phosphorylation analysis, modifications were enriched using TiO2 columns. We here demonstrate the efficiency of TE protocol for shotgun proteomics, providing similar results as urea-derived samples both at the qualitative and quantitative levels. The study of phosphorylation events is likewise compatible with TE, which actually results in a higher number of correctly localized sites than urea, while the nature of extracted sites appears somewhat distinct between both techniques. We thus conclude that our protocol is well suited for proteomics and as efficient as other more widely used workflows for mass spectrometry-based analysis.
2021-07-20 | PXD011971 | Pride
Project description:The potential of supercritical CO2 and ionic liquids (ILs) as alternatives to traditional extraction of natural compounds from plant material is of increasing importance. Both techniques offer several advantages over conventional extraction methods. These two alternatives have been separately employed on numerous ocassions, however, until now, they have never been combined for the extraction of secondary metabolites from natural sources, despite properties that complement each other perfectly. Herein, we present the first application of an IL-based dynamic supercritical CO2 extraction of six cannabinoids (CBD, CBDA, Δ9-THC, THCA, CBG and CBGA) from industrial hemp (Cannabis sativa L.). Various process parameters were optimized, i.e., IL-based pre-treatment time and pre-treatment temperature, as well as pressure and temperature during supercritical fluid extraction. In addition, the impact of different ILs on cannabinoid extraction yield was evaluated, namely, 1-ethyl-3-methylimidazolium acetate, choline acetate and 1-ethyl-3-methylimidazolium dimethylphosphate. This novel technique exhibits a synergistic effect that allows the solvent-free acquisition of cannabinoids from industrial hemp, avoiding further processing steps and the additional use of resources. The newly developed IL-based supercritical CO2 extraction results in high yields of the investigated cannabinoids, thus, demonstrating an effective and reliable alternative to established extraction methods. Ultimately, the ILs can be recycled to reduce costs and to improve the sustainability of the developed extraction process.
| S-EPMC8667783 | biostudies-literature
Project description:Four crosses were made between inbred Cannabis sativa plants with pure cannabidiol (CBD) and pure Delta-9-tetrahydrocannabinol (THC) chemotypes. All the plants belonging to the F(1)'s were analyzed by gas chromatography for cannabinoid composition and constantly found to have a mixed CBD-THC chemotype. Ten individual F(1) plants were self-fertilized, and 10 inbred F(2) offspring were collected and analyzed. In all cases, a segregation of the three chemotypes (pure CBD, mixed CBD-THC, and pure THC) fitting a 1:2:1 proportion was observed. The CBD/THC ratio was found to be significantly progeny specific and transmitted from each F(1) to the F(2)'s derived from it. A model involving one locus, B, with two alleles, B(D) and B(T), is proposed, with the two alleles being codominant. The mixed chemotypes are interpreted as due to the genotype B(D)/B(T) at the B locus, while the pure-chemotype plants are due to homozygosity at the B locus (either B(D)/B(D) or B(T)/B(T)). It is suggested that such codominance is due to the codification by the two alleles for different isoforms of the same synthase, having different specificity for the conversion of the common precursor cannabigerol into CBD or THC, respectively. The F(2) segregating groups were used in a bulk segregant analysis of the pooled DNAs for screening RAPD primers; three chemotype-associated markers are described, one of which has been transformed in a sequence-characterized amplified region (SCAR) marker and shows tight linkage to the chemotype and codominance.
| S-EPMC1462421 | biostudies-other