GC-TOF MS Research Articles

Comparative Metabolomics of High and Low Essential Oil Yielding Landraces of Betelvine (Piper betle L.) by Two-Dimensional Gas Chromatography Time-of-Flight Mass Spectrometry (GC × GC TOFMS)

Piper betle L. (Betelvine) is a highly important plant for its economical and medicinal value. However, its essential oil characterization has been limited to gas chromatography - mass spectrometry (GC-MS). In this study, the advanced two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC TOFMS) was used for the first time to separate and identify chemical constituents from leaf essential oils from two landraces of betelvine. A high essential oil yield (PB1) and a low essential oil yielding (PB13) were the landraces selected for GC × GC TOFMS analysis. Using GC × GC TOFMS, a total of 181 constituents were identified from PB1 and PB13 of which 86 chemical constituents were reported for the first time in the leaf essential oil. The total constituents were classified into sesquiterpene hydrocarbons (54 constituents), oxygenated sesquiterpenes (47 constituents), phenylpropanoids (14 constituents), oxygenated monoterpenes (14 constituents), and monoterpene hydrocarbons (13 constituents). On basis of area percentage, phenylpropanoids (38.81 to 30.90%) were present predominantly along with minor groups such as alcohols, esters, aldehydes, and alkanes. The major constituents were identified by GC × GC TOFMS as eugenol (9.73 to 13.20%), chavibetol acetate (7.40 to 5.12%), 4-ally-1,2-diacetoxybenzene (7.10 to 5.06%), eugenol acetate (3.46 to 2.47%), and chavibetol (3.24 to 2.39%). This study provides a detailed characterization of betelvine oil which can increase the pharmaceutical demands for novel drug discovery. Further, PB1 can be recommended for massive cultivation to increase the livelihood of farmers.

17 What’s in the Air We Breathe? Chemical Fingerprinting of Organic Species in Pm2.5 By Td-Gc×Gc-Tof Ms

Abstract The issue of urban air quality has received increased attention in recent years due to increased awareness of the potential health risks associated with particulate matter (PM) concentrations. Airborne particulate matter is composed of solid and/or liquid materials of various sizes, but the fine particles with diameters of <2.5 μm (so-called PM2.5) are of most concern, as long-term exposure to PM2.5 is thought to have the biggest impact on public health. PM2.5 consists of a wide range of components, from both primary emissions (e.g. vehicle exhausts or forest fires) and secondary reactions (i.e. compounds formed in the atmosphere by chemical reactions). It is important to have robust analysis of PM2.5 as the composition can give an indication of its source. PM2.5 is typically trapped onto quartz fibre filters and the semi-volatile organic compounds (SVOCs) are extracted using a solvent. Direct thermal desorption (TD) of the filter avoids this time-consuming sample preparation, for simple, yet sensitive, analysis of particulate matter. However, their identification and quantitation remains a challenge due to sample complexity. Analysts may attempt to address this using longer columns and/or slower oven temperature ramps, but this inevitably leads to longer analysis times. In recent years, the complexity of such samples has been revealed using the improved separation of comprehensive two-dimensional GC coupled with time-of-flight mass spectrometry (GC×GC–TOF MS). Here, we demonstrate the enhanced performance of a TD-GC×GC–TOF MS through analysis of a number of real-world PM2.5 filters.

Identification of 2,5-dimethyl-3-furanthiol in coffee by two dimensional GC TOF MS

Identification of 2,5-dimethyl-3-furanthiol in coffee by two dimensional GC TOF MS

The ADEM2 project: early pathogenic mechanisms of preschool wheeze and a randomised controlled trial assessing the gain in health and cost-effectiveness by application of the breath test for the diagnosis of asthma in wheezing preschool children

BackgroundThe prevalence of asthma-like symptoms in preschool children is high. Despite numerous efforts, there still is no clinically available diagnostic tool to discriminate asthmatic children from children with transient wheeze at preschool age. This leads to potential overtreatment of children outgrowing their symptoms, and to potential undertreatment of children who turn out to have asthma. Our research group developed a breath test (using GC-tof–MS for VOC-analysis in exhaled breath) that is able to predict a diagnosis of asthma at preschool age. The ADEM2 study assesses the improvement in health gain and costs of care with the application of this breath test in wheezing preschool children.MethodsThis study is a combination of a multi-centre, parallel group, two arm, randomised controlled trial and a multi-centre longitudinal observational cohort study. The preschool children randomised into the treatment arm of the RCT receive a probability diagnosis (and corresponding treatment recommendations) of either asthma or transient wheeze based on the exhaled breath test. Children in the usual care arm do not receive a probability diagnosis. Participants are longitudinally followed up until the age of 6 years. The primary outcome is disease control after 1 and 2 years of follow-up. Participants of the RCT, together with a group of healthy preschool children, also contribute to the parallel observational cohort study developed to assess the validity of alternative VOC-sensing techniques and to explore numerous other potential discriminating biological parameters (such as allergic sensitisation, immunological markers, epigenetics, transcriptomics, microbiomics) and the subsequent identification of underlying disease pathways and relation to the discriminative VOCs in exhaled breath.DiscussionThe potential societal and clinical impact of the diagnostic tool for wheezing preschool children is substantial. By means of the breath test, it will become possible to deliver customized and high qualitative care to the large group of vulnerable preschool children with asthma-like symptoms. By applying a multi-omics approach to an extensive set of biological parameters we aim to explore (new) pathogenic mechanisms in the early development of asthma, creating potentially interesting targets for the development of new therapies.Trial registrationNetherlands Trial Register, NL7336, Date registered 11–10-2018.

Biostimulation of Salicornia europaea L. crops with plant growth-promoting bacteria in laboratory and field conditions: effects on growth and metabolite profile.

The objective of the work was to assess the effect of biostimulation with selected plant growth-promoting bacteria on growth and metabolite profile of Salicornia europaea. Salicornia europaea seeds were inoculated with different combinations of plant growth-promoting bacteria Brevibacterium casei EB3, Pseudomonas oryzihabitans RL18, and Bacillus aryabhattai SP20. Plants germinated from inoculated seeds were grown either in laboratory conditions or in a saline crop field. Fresh and dry weight were determined at the end of the experiment, for biomass quantification. The microbiological quality of fresh shoots for human consumption as salad greens was assessed, and the persistence of the inoculated strains in the plant rhizosphere was confirmed by next-generation sequencing (Illumina) of the 16S rDNA gene. The primary metabolite profile of biostimulated plants was characterized by GC-TOF-MS.In laboratory conditions, inoculation with the two strains Br. casei EB3 and Ps. oryzihabitans RL18 caused the most significant increase in biomass production (fresh and dry weight), and caused a shift in the central metabolic pathways of inoculated plants toward amino acid biosynthesis. In the field experiment, no significant biostimulation effect was detected with any of the tested inoculants. Seed inoculation had no significant effect on the microbiological quality of the edible parts. The persistence of inoculants was confirmed in both experiments. Manipulation of the plant microbiome can trigger primary metabolic reconfiguration and modulate the plant metabolism while promoting plant growth.

Integrative metabolomics of plasma and PBMCs identifies distinctive metabolic signatures in Behçet’s disease

BackgroundBehçet’s disease (BD) is a systemic inflammatory disease that involves various organs. The clinical manifestation-based diagnosis of BD is a time-consuming process, which makes it difficult to distinguish from patients with similar symptoms. Moreover, an authentic biomarker has not been developed for accurate diagnosis yet. Our current study investigated the unique metabolic signatures of BD and explored biomarkers for precise diagnosis based on an untargeted metabolomic approach.MethodsIntegrative metabolomic and lipidomic profiling was performed on plasma samples of BD patients (n = 40), healthy controls (HCs, n = 18), and disease controls (DCs, n = 17) using GC-TOF MS and LC-Orbitrap MS. Additionally, the lipid profiles of 66 peripheral blood mononuclear cells (PBMCs) were analyzed from 29 BD patients, 18 HCs, and 19 DCs.ResultsPlasma metabolic dysfunction in BD was determined in carbohydrate, hydroxy fatty acid, and polyunsaturated fatty acid metabolisms. A plasma biomarker panel with 13 compounds was constructed, which simultaneously distinguished BD from HC and DC (AUCs ranged from 0.810 to 0.966). Dysregulated PBMC metabolome was signatured by a significant elevation in lysophosphatidylcholines (LPCs) and ether-linked lysophosphatidylethanolamines (EtherLPEs). Ten PBMC-derived lipid composites showed good discrimination power (AUCs ranged from 0.900 to 0.973). Correlation analysis revealed a potential association between disease activity and the metabolites of plasma and PBMC, including sphingosine-1 phosphate and EtherLPE 18:2.ConclusionsWe identified metabolic biomarkers from plasma PBMC, which selectively discriminated BD from healthy control and patients with similar symptoms (recurrent mouth ulcers with/without genital ulcers). The strong correlation was determined between the BD activity and the lipid molecules. These findings may lead to the development for diagnostic and prognostic biomarkers based on a better understanding of the BD pathomechanism.

Comprehensive profiling of controlled substances using gas chromatography coupled with time-of-flight mass spectrometry

Background: Drug profiling is performed to prove the presence of any controlled substances and to compare sample purity, to determine whether two samples are from the same origin, as well as to assess the expected monetary value of the drug samples. However, conventional analyses by gas chromatography – mass spectrometry (GC-MS) often lacks the sensitivity required to provide a full chemical fingerprint of the trace contaminants and cutting agents. Here, we apply gas chromatography coupled with time-of-flight mass spectrometry (GC-TOF MS) to identify trace differences between seized drug samples. Material and Methods: Six seized samples of suspected drugs of abuse, were prepared by dissolving of each drug powder in methanol, prior to analysis by GC–TOF MS. Resulting chromatograms were integrated, identified and compared using novel comparative analysis tools to determine trends and differences between the seized samples. Result: All six seized samples were found to contain diacetylmorphine (heroin), however the trace cutting agents and contaminants present varied significantly. For example, the presence of the common cutting agents, caffeine and acetaminophen (or paracetamol) was only noted for some of the samples, while others contained trace impurities, such as meconin and 6-acetylcodeine, which can be useful in linking samples to the same origin. Conclusion: Comprehensive profiling of seized drug samples was obtained using GC–TOF MS to identify not only the controlled substances, but also the various cutting agents and trace impurities present.

Comparative evaluation of physical characteristics and volatile flavor components of Bangia fusco-purpurea subjected to hot air drying and vacuum freeze-drying

Bangia fusco-purpurea is an economically important seaweed with Fujian characteristics. Given that its harvest is seasonal, drying is often used to remove moisture, extend storage time, and facilitate further processing. Hence, the current study sought to explore the impact of different drying processes on the quality and volatile fingerprints of Bangia fusco-purpurea. To this end, the effects of hot air drying (HAD) and vacuum freeze drying (VFD) on the drying characteristics, microstructure, rehydration, and volatile components of dried B. fusco-purpurea were investigated. The results showed that the water removal efficiency of HAD was significantly higher than that of VFD. However, VFD better preserved the skeletal structure of B. fusco-purpurea than HAD, with a faster rehydration rate and a more uniform cell structure after rehydration. Using electronic nose and comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOF MS), significant differences in the volatile profiles of fresh, HAD, and VFD B. fusco-purpurea were assessed. E-nose analysis revealed that both HAD and VFD treatments significantly reduced sulfides, aromatic compounds, and nitrogen oxides in fresh B. fusco-purpurea. However, the alcohol, aldehyde, and ketone contents were lower in the VFD samples compared with HAD and fresh samples, whereas the content of methyl flavor substances was significantly higher. GC × GC-TOF MS analysis revealed that the most abundant volatile categories in HAD and VFD were hydrocarbons, alcohols, and esters. The number of volatile components in the HAD samples was significantly lower than in the VFD and fresh samples. As drying progressed, hydrocarbons and alcohols were formed in dried B. fusco-purpurea due to the thermal degradation of carbohydrates, lipids, amino acids, and the Maillard reaction. There were also significant flavor differences between HAD and VFD B. fusco-purpurea. Thus, although HAD exhibits better drying efficiency, VFD has more significant advantages in terms of product quality.

Metabolome-Wide Reprogramming Modulated by Wnt/β-Catenin Signaling Pathway.

A family of signal transduction pathways known as wingless type (Wnt) signaling pathways is essential to developmental processes like cell division and proliferation. Mutation in Wnt signaling results in a variety of diseases, including cancers of the breast, colon, and skin, metabolic disease, and neurodegenerative disease; thus, the Wnt signaling pathways have been attractive targets for disease treatment. However, the complicatedness and large involveness of the pathway often hampers pinpointing the specific targets of the metabolic process. In our current study, we investigated the differential metabolic regulation by the overexpression of the Wnt signaling pathway in a timely-resolved manner by applying high-throughput and un-targeted metabolite profiling. We have detected and annotated 321 metabolite peaks from a total of 36 human embryonic kidney (HEK) 293 cells using GC-TOF MS and LC-Orbitrap MS. The un-targeted metabolomic analysis identified the radical reprogramming of a range of central carbon/nitrogen metabolism pathways, including glycolysis, TCA cycle, and glutaminolysis, and fatty acid pathways. The investigation, combined with targeted mRNA profiles, elucidated an explicit understanding of activated fatty acid metabolism (β-oxidation and biosynthesis). The findings proposed detailed mechanistic biochemical dynamics in response to Wnt-driven metabolic changes, which may help design precise therapeutic targets for Wnt-related diseases.

Lipid Serum Profiling of Boar-Tainted and Untainted Pigs Using GC×GC-TOFMS: An Exploratory Study.

Mass spectrometry (MS)-based techniques, including liquid chromatography coupling, shotgun lipidomics, MS imaging, and ion mobility, are widely used to analyze lipids. However, with enhanced separation capacity and an optimized chemical derivatization approach, comprehensive two-dimensional gas chromatography (GC×GC) can be a powerful tool to investigate some groups of small lipids in the framework of lipidomics. This study describes the optimization of a dedicated two-stage derivatization and extraction process to analyze different saturated and unsaturated fatty acids in plasma by two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS) using a full factorial design. The optimized condition has a composite desirability of 0.9159. This optimized sample preparation and chromatographic condition were implemented to differentiate between positive (BT) and negative (UT) boar-tainted pigs based on fatty acid profiling in pig serum using GC×GC-TOFMS. A chemometric screening, including unsupervised (PCA, HCA) and supervised analysis (PLS-DA), as well as univariate analysis (volcano plot), was performed. The results suggested that the concentration of PUFA ω-6 and cholesterol derivatives were significantly increased in BT pigs, whereas SFA and PUFA ω-3 concentrations were increased in UT pigs. The metabolic pathway and quantitative enrichment analysis suggest the significant involvement of linolenic acid metabolism.

Characterization of different meat flavor compounds in Guangdong small-ear spotted and Yorkshire pork using two-dimensional gas chromatography–time-of-flight mass spectrometry and multi-omics

In China, indigenous pigs are popular due to their excellent meat quality and unique flavor. In this study, the unique flavor was investigated using Guangdong small-ear spotted pig (GD) and their hybrid progeny, which produced high-end pork in the Chinese market. Two-dimensional gas chromatography-time-of-flight mass spectrometry (GC × GC-TOF MS) was used to analyze and quantify volatile organic compounds (VOCs) in the two pork types for identifying the unique flavor of these very popular pigs. The flavor is imparted by alkanes, aldehydes, ketones, and ethers. Flavor imparting compounds mainly include methyl isobutyl ketone, octane, pentane, and 3-ethyl-2, 2-dimethylheptane. High levels of volatilization of such compounds generated ethereal, acetonic, sweet, and green odors in the pork of GD. Hybrid offspring of Chinese and Western pig breeds can also inherit such special aromas, which are sometimes even more intense, with increased meat yield. Multi-omics analysis of the transcriptome and metabolome showed that AMD1 (Adenosylmethionine Decarboxylase 1) affects the p-cresol content by regulating the polyamine pathway, affecting the unique flavor of GD. This study revealed the unique flavor composition of GD, analyzed the advantages of species hybridization, and thus, provides a new direction and theoretical basis for pork quality improvement.

Comparison of PSA to Moringa Oleifera Seed Protein as Sorbent in QuEChERS: A Response Surface Methodology Optimization for Extraction of Some Endocrine Disrupting Chemicals in Food

This work aimed at optimizing the QuEChERS method with PSA and then comparing it with Moringa Oleifera seed protein as a clean-up sorbent for the extraction of endocrine-disrupting chemicals. The response surface methodology approach was used in the optimization. A design of experiment (DoE) was used to investigate the effect of the sample mass (0.5–3 g), centrifuge speed (3400–4000 rpm) and time (5–20 min), the mass of N a C l and MgSO 4 1 − 3 g , and solvent extraction volume (5–10 mL). The analysis was done using GC-ECD and GC × GC TOFMS. The PSA method which was later replaced with Moringa Oleifera seed protein presented optimal values of 3 g of sample, 150 mg PSA, 4000 rpm for 6 min centrifuge conditions, including 2 g NaCl and 2 g MgSO 4 extracted in 10 mL methanol, respectively. Moringa Oleifera seed protein gave better selectivity, and the detection limits ranged between 0.16 and 1.77 μ g kg − 1 with RSD values ≤ 13.32 % , respectively. Moreover, recoveries were between 76.2 ± 0.85% and 105.2 ± 2.24%. Application of the developed method in food samples detected some EDCs. This study has shown that Moringa Oleifera seed protein is a promising alternative to PSA in the clean-up of food-related samples using the QuEChERS approach.

Heterosis for capsacinoids accumulation in chili pepper hybrids is dependent on parent-of-origin effect

Heterosis for agronomic traits is a widespread phenomenon that underpins hybrid crop breeding. However, heterosis at the level of cellular metabolites has not yet been fully explored. Some metabolites are highly sought after, like capsaicinoids found in peppers of the Capsicum genus, which confer the characteristic pungent (‘hot’) flavour of the fruits. We analysed the metabolic profile of the fruit placenta and pericarp of inter- and intra-specific hybrids of two species of Capsicum peppers, C. chinense (cv. Habanero and cv. Biquinho) and C. annuum var. annuum (cv. Jalapeño and cv. Cascadura Ikeda) in complete diallel crosses with reciprocals. The parents and hybrids were grown in a glasshouse and the profile of primary metabolites (sugars, amino acids and organic acids) and capsaicinoids was generated via gas chromatography–time of flight-mass spectrometry (GC–TOF-MS) and ultra-performance liquid chromatography coupled to a mass spectrometer (UPLC-MS), respectively. We found considerable heterotic effects specifically for capsaicinoids accumulation in the fruit placenta of the hybrids, including those derived from non-pungent parents. Furthermore, a large fraction of fruit primary metabolism was influenced by the specific cross combination, with marked parent-of-origin effects, i.e. whether a specific genotype was used as the pistillate or pollen parent. The differences in metabolite levels between the hybrids and their parents provide a snapshot of heterosis for primary and secondary metabolites and may contribute to explain the manifestation of whole-plant heterotic phenotypes.

Analysis of VOCs in Urine Samples Directed towards of Bladder Cancer Detection.

Bladder cancer is one of most common types of cancer diagnosed in the genitourinary tract. Typical tests are costly and characterized by low sensitivity, which contributes to a growing interest in volatile biomarkers. Head space solid phase microextraction (SPME) was applied for the extraction of volatile organic compounds from urine samples, and gas chromatography time of flight mass spectrometry (GC×GC TOF MS) was used for the separation and detection of urinary volatiles. A cohort of 40 adult patients with bladder cancer and 57 healthy persons was recruited. Different VOC profiles were obtained for urine samples taken from each group. Twelvecompounds were found only in the samples from theBC group.The proposed candidate biomarkers are butyrolactone; 2-methoxyphenol; 3-methoxy-5-methylphenol; 1-(2,6,6-trimethylcyclohexa-1,3-dien-1-yl)-2-buten-1-one; nootkatone and 1-(2,6,6-trimethyl-1-cyclohexenyl)-2-buten-1-one.Since most of the studies published in the field are proving the potential of VOCs detected in urine samples for the screening and discrimination of patients with bladder cancer from healthy, but rarely presenting the identity of proposed biomarkers, our study represents a novel approach.

Emissions of organic compounds from western US wildfires and their near-fire transformations

Abstract. The size and frequency of wildfires in the western United States have been increasing, and this trend is projected to continue, with increasing adverse consequences for human health. Gas- and particle-phase organic compounds are the main components of wildfire emissions. Some of the directly emitted compounds are hazardous air pollutants, while others can react with oxidants to form secondary air pollutants such as ozone and secondary organic aerosol (SOA). Further, compounds emitted in the particle phase can volatize during smoke transport and can then serve as precursors for SOA. The extent of pollutant formation from wildfire emissions is dependent in part on the speciation of organic compounds. The most detailed speciation of organic compounds has been achieved in laboratory studies, though recent field campaigns are leading to an increase in such measurements in the field. In this study, we identified and quantified hundreds of gas- and particle-phase organic compounds emitted from conifer-dominated wildfires in the western US, using two two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC × GC ToF-MS) instruments. Observed emission factors (EFs) and emission ratios are reported for four wildfires. As has been demonstrated previously, modified combustion efficiency (MCE) was a good predictor of particle-phase EFs (e.g., R2=0.78 and 0.84 for sugars and terpenoids, respectively), except for elemental carbon. Higher emissions of diterpenoids, resin acids, and monoterpenes were observed in the field relative to laboratory studies, likely due to distillation from unburned heated vegetation, which may be underrepresented in laboratory studies. These diterpenoids and resin acids accounted for up to 45 % of total quantified organic aerosol, higher than the contribution from sugar and sugar derivatives. The low volatility of resin acids makes them ideal markers for conifer fire smoke. The speciated measurements also show that evaporation of semi-volatile organic compounds took place in smoke plumes, which suggests that the evaporated primary organic aerosol can be a precursor of SOAs in wildfire smoke plumes.

Qualitative characterisation and identification of microplastics in a freshwater dam at Gauteng Province, South Africa, using pyrolysis-gas chromatography-time of flight-mass spectrometry (Py-GC-ToF-MS).

Pyrolysis GC-ToF-MS-based analytical study was employed in the identification of microplastics (MPs) in the freshwater of a dam Rietvlei (RTV) located at Gauteng Province, South Africa. These MPs extracted in five locations of the dam were found to contain five different plastic polymeric constituents including PE, PS, PA, PVC and PET along with phthalate esters and fatty acid (amides and esters) derivatives as additives. Based on the fragmented pyrolyzate products, the contribution of plastic polymers and additives was 74% and 26% respectively. Among polymers, PA was dominant with 52% followed by PVC (16%) and others (13%) such as PE, PET and PS in MPs. Scanning electron micrographs of MPs in this aquatic body displayed the rough and fibrous typed patterns. The residual mass of 8-14% was left after the thermal degradation of MPs in RTV samples in the temperature range of 500-550°C. The results of thermogravimetry (TGA) and energy-dispersive (EDS) analyses are mutually dependent and coherent to each other by way of demonstrating the presence of various inorganic compounds in the form of additives and/or sorbates. The lessened intensities of carbonyl stretching in PA (1625cm-1) and PET (1725cm-1) type of MPs attributed the occurrence of degradation and weathering in this aquatic system. The possible causes to the contamination of MPs in this freshwater are the located industries and poor waste management strategies being practised in this densely populated city. Based on the industry, waste management and population perspectives, the increased contamination of MPs is very likely in this freshwater which will drastically affect the ecosystem in the near future. Based on the characterisation results, the presence of various polymers, additives and the metals in MPs is envisaged to deteriorate the aquatic life along with successive risks for the people as a consequence of bio-magnification.

Soil microbiome and metabolome analysis reveals beneficial effects of ginseng–celandine rotation on the rhizosphere soil of ginseng-used fields

Ginseng (Panax ginseng C.A.Meyer) cultivation is hindered by the continuous cropping obstacle, whereby without appropriate rotations yield decline and fields must be abandoned. Organism interactions in the rhizosphere alleviate and improve the obstacles to continuous cropping of ginseng. In this study, high-throughput sequencing and gas chromatography coupled with time-of-flight mass spectrometry (GC–TOF-MS) in combination with analysis of soil chemical properties were used to investigate soil microecological changes after 5 years of celandine (Chelidonium majus L.) cultivation in land previously planted with ginseng for 6 years. Comparison of data for the ginseng–celandine rotation (experimental group) and ginseng–fallow (control group) revealed that the celandine rhizosphere TOC, AN, and AK contents as well as PPO activity and Cat activity were significantly increased. Rotation increased the abundance of some bacteria and fungal in the rhizosphere of celandine rhizosphere soil, such as Arthrobacter, Metarhizium, and Exophiala. The relative abundance of Fusarium in ginseng–celandine rotation soil was lower than that of ginseng–fallow soil, which may effectively reduce the incidence of ginseng root rot. The non-targeted metabolites and microbial community structure of the rhizosphere soil were changed with celandine cultivation. In celandine rhizosphere soil, the abundance of the bacterial and fungal communities was positively or negatively correlated with rhizosphere soil metabolites. Pathway enrichment analysis showed that starch and sucrose metabolism were significantly enriched in the celandine rhizosphere soil, a result consistent with the results shown in bacterial community function prediction and non-targeted metabolomics. The present combined analysis of the soil microbiome and non-targeted metabolome provided insight into the effects and mechanism of ginseng–celandine rotation on the soil micro-ecosystem of ginseng-used fields. Briefly, crop rotation with celandine was beneficial to soil quality, and it is a good option to alleviate Ginseng continuous cropping obstacles.

Construction of surrogate fuels for lower freezing point diesels based on component and functional groups analysis

For the purpose of formulating surrogate fuels for two lower freezing point diesels, namely −35# diesel and − 50# diesel, two-dimensional gas chromatography with time-of-flight mass spectrometry (GC × GC–TOFMS) was utilized to analyze their components in detail. And nuclear magnetic resonance (NMR) spectroscopy was used to obtain the composition of functional groups in hydrocarbons, which were classified into 11 carbon types according to different structural properties of molecules. The GC × GC–TOFMS results reflected that −50# diesel contains less unsaturated hydrocarbons, while −35# diesel contains more heavy hydrocarbons. The NMR spectroscopy results indicated that −50# diesel contains more CH3 (primary carbon) and n-alkane CH2 (secondary carbon), and the H/C ratio reflected by the analysis results of NMR spectroscopy were consistent with the H/C ratio measured by ASTM D5291. Based on the results of GC × GC–TOFMS, one five-component and one seven-component surrogate fuels were constructed for −35# diesel and −50# diesel through genetic algorithm by matching the density, viscosity, cetane number, distillation curve and functional groups. Then spray penetration of the surrogate fuels were validated and the result are compared with the experimental results of lower freezing point diesel under spray condition (Pamb = 3 MPa, Tamb = 800 K). It turned out that the both the target properties and the non-target properties showed good agreement that the surrogate fuels can emulate properties of lower freezing point diesels well.

Abstract 5319: Treatment of K562 leukemia cells with an experimental UBE2K modifier identifies multi-omic changes associated with altered oncogenic processes

Abstract Ubiquitination is a conserved post translation modification involving covalent attachment of ubiquitin protein and is known to regulate many biological processes, including proteasomal degradation. Three major families of enzymes are involved in the regulation of ubiquitination, including activating enzyme E1, Ubiquitin conjugating enzyme E2 and ubiquitin ligase E3. UBE2K is an E2 conjugating ligase that was identified as an anti-cancer drug target from the BERG Interrogative Biology® platform, an artificial intelligence multi-omics analytical method employing Bayesian algorithms. Herein, we used proteomics, lipidomics and metabolomics to investigate the impact of the treatment of UBE2K small molecule ligand (BRG0451) on K562 leukemia cells. K562 cells were treated with 30, 100 and 300 nM concentrations for 24 hours with BRG0451 or Paclitaxel or 0.1% DMSO (Control). Cells were pelleted and analyzed using a multi-omics approach. Proteomic analysis was performed using Thermo Q-Exactive+ LC MS/MS analysis. Lipidomic analysis was performed using SCIEX TripleTOF MS/MS ALL shotgun workflow and metabolomics was performed using 3 different platforms (High resolution RP-LC-MS, HILIC QqQ LC-MS/MS and GC-TOF MS). Unsupervised clustering and differential analysis were used to investigate the impact of the treatments. Proteomic analysis identified and quantified 6930 proteins from K562 cells using TMT labelling with offline 24 fractions and LC-MS/MS. Structural lipidomics analysis evaluated 1980 lipid molecular species and metabolomics analysis identified over 700 metabolites using GC-MS, LC-MS and LC-MS/MS. Multiomics and regression analysis for 30 nM BRG0451 treatment revealed no distinct pattern of omics variables. However, treatment on K562 cells with 300 nM treatment demonstrated 97 differentially expressed proteins compared to control. Pathway analysis revealed chromatin remodeling, and more specifically, regulation of chromatin silencing and localization to nucleolus as major pathways impacted by differentially expressed proteins. Similar pathways were impacted by Paclitaxel and Nocodazole treatment compared to control. Additionally, metabolomic and lipidomic differentials were observed with 300 nM BRG0451 treatment. Structural lipidomics revealed dose -dependent changes in triacylglycerols and cholesterol esters, glycolipid monounsaturated species, and glycolipid medium carbon chain subgroups. Dose dependent impact on amino acids metabolism, purine metabolism, and pyrimidine metabolism was observed with a high degree of similarity for compared drugs. Herein, we demonstrated the use of multi-omics technology in deconvoluting the impact of BRG0451 on independent biological pathways, revealing the intricate mechanisms targeting cell cycle as well as ubiquitin regulator components in a leukemia cell line. Citation Format: Punit Shah, Richard Searfoss, Valerie Bussberg, Bennett Greenwood, Shraddha Karmacharya, Allison MacDonald, Kennedy Ofori-Mensa, Vladimir Tolstikov, Pragalath Sundararajan, Maria-Dorothea Nastke, Eric M. Grund, Gregory M. Miller, Stephane Gesta, Rangaprasad Sarangarajan, Elder Granger, Niven R. Narain, Vivek K. Vishnudas, Michael A. Kiebish. Treatment of K562 leukemia cells with an experimental UBE2K modifier identifies multi-omic changes associated with altered oncogenic processes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5319.

Leymus chinensis Adapts to Degraded Soil Environments by Changing Metabolic Pathways and Root Exudate Components.

Phytoremediation is a promising remediation strategy for degraded soil restoration. Root exudates are the main carrier substances for information communication and energy transfer between plant roots and soil, which play non-negligible roles in the restoration process. This work investigated the adaptation of Leymus chinensis root exudates to different degraded levels of soil and the mechanism of rhizosphere restoration in a 3-year degraded soil field study. We found that the soil quality at each degradation level significantly increased, with the soil organic matter (SOM) content slightly increasing by 1.82%, moderately increasing by 3.27%, and severely increasing by 3.59%, and there were significant increases in the contents of available nutrients such as available phosphorus (AP), ammonia nitrogen (AN), and nitrate nitrogen (NN). The physiological activities indicated that root tissue cells also mobilize oxidative stress to respond to the soil environment pressure. A total of 473 main components were obtained from root exudates by gas chromatography–time-of-flight mass spectrometry (GC–TOFMS), including acids, alcohols, carbohydrates, and other major primary metabolites. OPLS-DA revealed that soil degradation exerted an important influence on the metabolic characteristics of root exudates, and the numbers of both up- and downregulated metabolic characteristic peaks increased with the increase in the degree of degradation. Forty-three metabolites underwent clear changes, including some defense-related metabolites and osmotic adjustment substances that were significantly changed. These changes mainly mobilized a series of lipid metabolism pathways to maintain the fluidity of membrane function and help plants adapt to unfavorable soil environmental conditions. The PPP energy metabolism pathway was mobilized in response to slight degradation, and TCA energy pathways responded to the environmental pressure of severe soil degradation.