Impact of breath sample collection method and length of storage of breath samples in Tedlar bags on the level of selected volatiles assessed using gas chromatography-ion mobility spectrometry (GC-IMS).

The analysis of volatile organic compounds (VOCs) in exhaled air has attracted the interest of the scientific community because it provides the possibility of monitoring physiological and metabolic processes and non-invasive diagnostics of various diseases. However, this method remains underused in clinical practice as well as in research because of the lack of standardized procedures for the collection, storage and transport of breath samples, which would guarantee good reproducibility and comparability of results. The method of sampling, as well as the storage time of the breath samples in the polymer bags used for sample storage and transport, affect the composition and concentration of VOCs present in the breath samples. The aim of our study was to compare breath samples obtained using two methods with fully disposable equipment: a Haldane sampling tube intended for direct breath collection and breath samples exhaled into a transparent Tedlar bag. The second task was to monitor the stability of selected compounds of real breath samples stored in a Tedlar bag for 6 h. Gas chromatography coupled with ion mobility spectrometry (GC-IMS) implemented in the BreathSpec®device was used to analyse exhaled breath. Our results showed a significant difference in the signal intensity of some volatiles when taking a breath sample with a Haldane tube and a Tedlar bag. Due to its endogenous origin, acetone levels were significantly higher when the Haldane tube sampler was used while elevated levels of 2-propanol and unidentified VOC (designated as VOC 3) in the Tedlar bag samples likely originated from contamination of the Tedlar bags. The VOC stability study revealed compound-specific signal intensity changes of the selected VOCs with storage time in the Tedlar bags, with some volatiles showing increasing signal intensity during storage in Tedlar bags. This limits the use of Tedlar bags only for very limited time and carefully selected purpose. Our results highlight the importance of careful design and implementation of experiments and clinical protocols to obtain relevant and reliable results.

Optimized workflow for unknown screening using gas chromatography high-resolution mass spectrometry expands identification of contaminants in silicone personal passive samplers.

RATIONALE: Silicone wristbands have emerged as valuable passive samplers for monitoring of personal exposure to environmental contaminants in the rapidly developing field of exposomics. Once deployed, silicone wristbands collect and hold a wealth of chemical information that can be interrogated using high-resolution mass spectrometry (HRMS) to provide a broad coverage of chemical mixtures. METHODS: Gas chromatography coupled to Orbitrap™ mass spectrometry (GC/Orbitrap™ MS) was used to simultaneously perform suspect screening (using in-house database) and unknown screening (using vendor databases) of extracts from wristbands worn by volunteers. The goal of this study was to optimize a workflow that allows detection of low levels of priority pollutants, with high reliability. In this regard, a data processing workflow for GC/Orbitrap™ MS was developed using a mixture of 123 environmentally relevant standards consisting of pesticides, flame retardants, organophosphate esters, and polycyclic aromatic hydrocarbons as test compounds. RESULTS: The optimized unknown screening workflow using a search index threshold of 750 resulted in positive identification of 70 analytes in validation samples, and a reduction in the number of false positives by over 50%. An average of 26 compounds with high confidence identification, 7 level 1 compounds and 19 level 2 compounds, were observed in worn wristbands. The data were further analyzed via suspect screening and retrospective suspect screening to identify an additional 36 compounds. CONCLUSIONS: This study provides three important findings: (1) a clear evidence of the importance of sample cleanup in addressing complex sample matrices for unknown analysis, (2) a valuable workflow for the identification of unknown contaminants in silicone wristband samplers using electron ionization HRMS data, and (3) a novel application of GC/Orbitrap™ MS for the unknown analysis of organic contaminants that can be used in exposomics studies.

Improving Quantification of tabun, sarin, soman, cyclosarin, and sulfur mustard by focusing agents: A field portable gas chromatography-mass spectrometry study.

Commercial gas chromatograph-mass spectrometers, one of which being Inficon's HAPSITE® ER, have demonstrated chemical detection and identification of nerve agents (G-series) and blistering agents (mustard gas) in the field; however most analyses relies on self-contained or external calibration that inherently drifts over time. We describe an analytical approach that uses target-based thermal desorption standards, called focusing agents, to accurately calculate concentrations of chemical warfare agents that are analyzed by gas chromatograph-mass spectrometry. Here, we provide relative response factors of focusing agents (2-chloroethyl ethyl sulfide, diisopropyl fluorophosphate, diethyl methylphosphonate, diethyl malonate, methyl salicylate, and dichlorvos) that are used to quantify concentrations of tabun, sarin, soman, cyclosarin and sulfur mustard loaded on thermal desorption tubes (Tenax® TA). Aging effects of focusing agents are evaluated by monitoring deviations in quantification as thermal desorption tubes age in storage at room temperature and relative humidity. The addition of focusing agents improves the quantification of tabun, sarin, soman, cyclosarin and sulfur mustard that is analyzed within the same day as well as a 14-day period. Among the six focusing agents studied here, diisopropyl fluorophosphate has the best performance for nerve agents (G-series) and blistering agents (mustard gas) compared to other focusing agents in this work and is recommended for field use for quantification. The use of focusing agent in the field leads to more accurate and reliable quantification of Tabun (GA), Sarin (GB), Soman (GD), Cyclosarin (GF) and Sulfur Mustard (HD) than the traditional internal standard. Future improvements on the detection of chemical, biological, radiological, nuclear, and explosive materials (CBRNE) can be safely demonstrated with standards calibrated for harmful agents.

Distinguishing between long-term-stored and fresh chili pepper powder through fingerprinting of volatiles by headspace capillary-gas chromatography-ion mobility spectrometry.

Long-term storage of chili pepper powder results in physicochemical and microbiological changes that decrease its commercial value; these changes occur owing to fungal growth and production of off-flavor compounds. Herein, long-term-stored chili pepper powder (LSCPP) and fresh chili pepper powder (FCPP) were analyzed using internal transcribed spacer sequencing and volatile organic compound fingerprinting by headspace capillary-gas chromatography-ion mobility spectrometry. Fungal analysis detected only Xeromyces bisporus with high accuracy in all the analyzed LSCPP samples. However, the proliferation of X. bisporus on nonspecific spots complicated the distinguishing process between the two groups based solely on fungal analysis. Therefore, nine compounds (three ketones, one alcohol, two aldehydes, one ester, one furan, and one sulfur compound) obtained by autoxidation and fungal metabolism were selected as potential markers for distinguishing LSCPP and FCPP. These above-mentioned substances, which were confirmed as off-flavor species owing to "stale" odor, emitted lipid fragrance and were used to successfully distinguish LSCPP from FCPP using principal component analysis and linear discriminant analysis. PRACTICAL APPLICATION: According to the research results, it was possible to discriminate between long-term stored and fresh chili pepper powders using nine VOC markers for quality control in industry. In addition, the fungus generated from long-term storage of chili pepper powder was Xeromyces bisporus, which was confirmed to be safe for intake because it does not form secondary toxic metabolites.

Total-transfer comprehensive three-dimensional gas chromatography with time-of-flight mass spectrometry.

Although comprehensive two-dimensional (2D) gas chromatography (GC × GC) is a powerful technique for complex samples, component overlap remains likely. An intriguing route to address this challenge is to utilize the additional peak capacity and chemical selectivity provided by comprehensive three-dimensional (3D) gas chromatography (GC3), especially with time-of-flight mass spectrometry detection (GC3-TOFMS). However, the GC3-TOFMS instrumentation reported to date has employed one or both modulators with a duty cycle < 100%, making the potential gain in detection sensitivity over GC × GC modest, or perhaps even worse. Herein, we describe instrumentation for GC3-TOFMS in which both modulators provide total-transfer (100% duty cycle). Specifically, the instrument is based on the facile modification of a commercial thermally modulated comprehensive GC × GC-TOFMS platform for modulation from the 1D column to the 2D column, with recently described dynamic pressure gradient modulation (DPGM) as the second modulator from the 2D column to the 3D column, which is a total-transfer flow modulation technique. Area measurements of 1D peaks are compared to the sum of 3D peak areas to validate the assumption that total-transfer from 1D to 3D is accomplished. Additionally, peak heights were amplified by as high as a factor of 177 (x̅ = 130, s = 47) via comparison of 1D peak heights to the maximum 3D peak heights. Column selection is explored, with emphasis on the resulting peak width-at-base on each dimension and usage of 3D space as evaluation metrics. Using a nonpolar × polar × ionic liquid column combination, an effective peak capacity which considers modulation-induced broadening as high as 32,300 for select analytes was achieved (x̅ = 19,900, s = 10,700). The analytical benefits of employing three selective phases, mass spectrometry detection, and total-transfer modulation are explored with separations of a metabolomics-type sample, i.e., derivatized porcine serum, and a jet fuel spiked with various sulfur-containing compounds.

Dissecting Sesquiterpene Profiles of Lemberger Red Wines Using Ex Vivo Tissue Deuterium-Labeling and Comprehensive Two-Dimensional Gas Chromatography-Time-of-Flight-Mass Spectrometry.

By means of ex vivo tissue deuterium-labeling using the stable isotope-labeled precursor [6,6,6-2H3]-(±)-mevalonolactone and microvinification experiments, we were able to show for the first time that the three sesquiterpene hydrocarbons, guaiazulene, δ-selinene, and selina-3,7(11)-diene, in Lemberger red wines do not originate from acid-catalyzed cyclization of yeast-derived farnesol and nerolidol. The three aforementioned sesquiterpene hydrocarbons could be unambiguously identified as grape-derived secondary metabolites and can therefore be considered as variety-specific marker compounds. The analysis of sesquiterpene hydrocarbons in red wine samples was performed by solid-phase extraction-headspace solid-phase microextraction-comprehensive two-dimensional gas chromatography-time of flight-mass spectrometry. The developed methodology paves the way for an analytical verification of grape variety labeling in wine authenticity control.

A fast and selective gas liquid microextraction of semiochemicals for quantitative analysis in plants.

A trapping-based gas liquid microextraction (GLME) method coupled with gas chromatography-mass spectrometry (GC-MS) was utilized to qualitatively and quantitatively characterize semiochemicals in plants. The main GLME extraction efficiency associated parameters (heating temperature and extraction time) were optimized. The results obtained from GLME process were compared with those of steam distillation and ultrasonic extraction, and the recovery, peak number and reproducibility were evaluated by using Thuja koraiensis Nakai as a representative plant. Furthermore, the quantitative performances of the GLME in terms of sample amount, recoveries of spiked standards and correlation were systematically evaluated using standard addition method, which gave a good quantitative ability for all the compounds with squares of correlation coefficient (r2) of higher than 0.99. Finally, the contents of α-pinene, camphene, linalool, α-terpinenol, ß-caryophyllene, α-caryophyllene, and totarol in Thuja koraiensis Nakai samples were quantified, and their concentrations (SD, n = 3) were; 0.65 (0.06), 0.62 (0.05), 4.12 (0.15), 0.99 (0.08), 1.11 (0.07), 0.63 (0.04), and 21.91 (0.25) µg g-1, respectively. It was demonstrated that GLME is a powerful sample preparation technique for quantitative and qualitative analysis of plant semiochemicals.

Assessment of a portable quadrupole-based gas chromatography mass spectrometry for seized drug analysis.

The cutting agents, classified as diluents (pharmacologically inactive) or adulterants (pharmacologically active), are substances commonly used to cut drugs of abuse to increase profits. These substances are constantly changing over time, increasing the risks to the user's health caused by the compounds' potential individual toxicities as well as their drug-drug interactions. This work aimed to develop and validate a screening method using a portable quadrupole-based gas chromatography mass spectrometer (FLIR Griffin™ G510) to identify drugs of abuse and adulterants in seized material, and compare it with a well validated standard technology, gas chromatography mass spectrometry (GC-MS). The method was validated for the identification of alprazolam, amphetamine, aminopyrine, benzocaine, caffeine, cocaine, codeine, diltiazem, ephedrine, fentanyl, fenethylline, furanylfentanyl, heroin, hydroxyzine, levamisole, lidocaine, methamphetamine, morphine, noramidopyrine (a marker of metamizole), phencyclidine, phenacetin, procaine, strychnine and xylazine. The targeted substances were chosen based on current intelligence regarding prevalent adulterants observed in multiple jurisdictions. Interference, precision, robustness and carryover were evaluated. The method was successfully validated and proved to be suitable to detect and identify the 24 target compounds proposed. The reliability of the instrument for detecting the presence of targeted compounds was analyzed by using Receiver Operating Characteristic (ROC) analysis. The portable quadrupole-based gas chromatography mass spectrometer was considered suitable for use in forensic analysis as a screening method.

Quantitative Determination of H2 in Human Blood by 22Ne-aided Gas Chromatography-Mass Spectrometry Using a Single Quadrupole Instrument.

Here, we present a quantitative method for H2 detection by gas chromatography-selected ion monitoring-mass spectrometry (GC-SIM-MS) using a single quadrupole instrument. Additionally, the developed method was applied to the detection of H2 in human blood by GC-SIM-MS analysis using the existing 22Ne in air as an internal standard (IS). H2 was analyzed by GC-SIM-MS using a single quadrupole instrument with double TC-Molsieve 5A capillary columns for the separation of permanent gases. The detections of H2 (analyte) and 22Ne (IS) were performed at m/z 2 and 22, respectively, by GC-SIM-MS. The analyte and IS were separated using He as the carrier gas. The ratio of the peak area of H2 to 22Ne was employed to obtain a calibration curve for H2 determination in the gas phase. The proposed GC-SIM-MS method exhibited high sensitivity in terms of the limits of detection (LOD) (1.7 ppm) and quantitation (LOQ) (5.8 ppm) for H2 analysis. The developed quantitative assay of H2 in the headspace of blood samples achieved high repeatability with a relative standard deviation (RSD) of 1.4 - 4.7%. We successfully detected and quantified H2 in the headspaces of vacuum blood-collection tubes containing whole blood from 11 deceased individuals with several causes of death by employing the developed GC-SIM-MS method. The quantitative value of H2 ranged from 5 to 905 ppm. The proposed GC-SIM-MS method was applicable to the quantitative assay of H2 in biological samples without tedious pretreatment requirements.

Online Scavenging of Trace Analytes in Complex Matrices for Fast Analysis by Carbon Dioxide Bubbling Extraction Coupled with Gas Chromatography-Mass Spectrometry.

Carbon dioxide (CO2) microbubbles can selectively enrich organic solutes from sea spray aerosols. Common bubbling extractions are normally followed by off-line separation/detection through methods such as mass spectrometry, chromatography, and spectroscopy. However, it is necessary to establish extractions with online separation and identification systems to improve efficiency and minimize sample loss. In this study, CO2 is used to form microbubbles in the sample solution, and trace analytes in the solution are transported to the gas phase by bubble bursting. Analytes at the liquid-gas interface are directly released into the trapping device, followed by thermal desorption for gas chromatography-mass spectrometry. For polycyclic aromatic hydrocarbons, the dependence of the extraction efficiency on various parameters has been analyzed. The method reported here provides high efficiency and minimizes the loss of trace volatiles with a better signal strength and signal-to-noise ratio than other gases. These features make the proposed method a rapid method to detect and quantify volatile/semivolatile analytes in complex liquid matrices. In addition to the preconcentration of organics, metal ions, and inorganic anions, a noticeable decrease of metal-organic compounds in the aqueous solution was shown for the first time. We finally propose a simple model of chemical partitioning in CO2 bubbling extraction of liquid samples for guiding online monitoring of trace analytes in real-world samples.

Chemical composition, antioxidant, anti-inflammatory and antiproliferative activities of the essential oil of Cymbopogon nardus, a plant used in traditional medicine.

Objectives Natural products commonly used in traditional medicine, such as essential oils (EOs), are attractive sources for the development of molecules with anti-proliferative activities for future treatment of human cancers, e.g., prostate and cervical cancer. In this study, the chemical composition of the EO from Cymbopogon nardus was characterized, as well as its antioxidativeproperties and anti-inflammatory and antiproliferative activities on LNCaP cells derived from prostate cancer. Methods The chemical composition of the EO was determined by GC/FID and GC/MS analyses. The antioxidative properties were assessed using DPPH radical scavenging assay and ABTS+• radical cation decolorization assay, and the anti-inflammatory capacity was determined by the inhibition of the lipoxygenase activity. Antiproliferative activity was evaluated by MTT assay. Results Collectively, our data show that the major constituents of C. nardus EO are citronellal (33.06 %), geraniol (28.40 %), nerol (10.94 %), elemol (5.25 %) and delta-elemene (4.09 %). C. nardus EO shows modest antioxidant and anti-inflammatory activity compared to the standard galic acid. C. nardus EO exhibits the best antiproliferative activity on the prostate cancer cell line LNCaP with an IC50 of 58.0 ± 7.9 µg/mL, acting through the induction of the cell cycle arrest. Conclusions This study has determined that C. nardus EO efficiently triggers cytotoxicity and pens a new field of investigation regarding the putative use of this EO in vivo.

The intelligent knife (iKnife) and its intraoperative diagnostic advantage for the treatment of cervical disease.

Clearance of surgical margins in cervical cancer prevents the need for adjuvant chemoradiation and allows fertility preservation. In this study, we determined the capacity of the rapid evaporative ionization mass spectrometry (REIMS), also known as intelligent knife (iKnife), to discriminate between healthy, preinvasive, and invasive cervical tissue. Cervical tissue samples were collected from women with healthy, human papilloma virus (HPV) ± cervical intraepithelial neoplasia (CIN), or cervical cancer. A handheld diathermy device generated surgical aerosol, which was transferred into a mass spectrometer for subsequent chemical analysis. Combination of principal component and linear discriminant analysis and least absolute shrinkage and selection operator was employed to study the spectral differences between groups. Significance of discriminatory m/z features was tested using univariate statistics and tandem MS performed to elucidate the structure of the significant peaks allowing separation of the two classes. We analyzed 87 samples (normal = 16, HPV ± CIN = 50, cancer = 21 patients). The iKnife discriminated with 100% accuracy normal (100%) vs. HPV ± CIN (100%) vs. cancer (100%) when compared to histology as the gold standard. When comparing normal vs. cancer samples, the accuracy was 100% with a sensitivity of 100% (95% CI 83.9 to 100) and specificity 100% (79.4 to 100). Univariate analysis revealed significant MS peaks in the cancer-to-normal separation belonging to various classes of complex lipids. The iKnife discriminates healthy from premalignant and invasive cervical lesions with high accuracy and can improve oncological outcomes and fertility preservation of women treated surgically for cervical cancer. Larger in vivo research cohorts are required to validate these findings.

Minor compounds and potential interferents in gas chromatographic analyses of human serum fatty acids.

Fatty acids from 100 randomly selected human serum samples were esterified to fatty acid methyl esters and analyzed by gas chromatography with flame ionization detector. A subset of the 20 samples that spans the variation in the original set of 100 samples were thereafter analyzed by gas chromatography-mass spectrometry (GC-MS). The GC-MS data were acquired using capillary columns with two different stationary phases, BP20 (polyethylene glycol) and BPX70 (cyanopropyl polysilphenylene-siloxane). Equivalent chain lengths on the two columns are reported for 69 compounds that constituted more than 0.1% of the chromatographic area in at least one sample. Of these, 39 compounds were identified as regular fatty acid methyl esters. The remaining 30 compounds were decomposition products from cholesterol, dimethylacetals, three compounds that have been linked to poor kidney function, and 13 compounds that are currently unidentified. The retention index patterns showed that on both columns there were 16 compounds that were separated by less than 0.05 equivalent chain length units from the nearest neighbor, meaning that they were overlapping or poorly resolved. The relationship between the peak threshold level and the number of peaks found above the level predicts a dramatic increase in the number of peaks that have to be resolved if the threshold is lowered below 0.1%.

Determination and comparison of agarwood from different origins by comprehensive two-dimensional gas chromatography-quadrupole time-of-flight mass spectrometry.

Agarwood, a species of resinous heartwood, is a precious medicinal plant and a type of rare natural spice, which is widely used in medicine, cosmetics, religious activities, and other fields. In this study, agarwood samples from eight different regions across four countries were analyzed by comprehensive two-dimensional gas chromatography-quadrupole time-of-flight mass spectrometry. A total of 232 species were identified (the match factors of these compounds were above 750). The main compounds of agarwood are oxygenated sesquiterpenes and chromones. The compositions of India1 and Malaysia2 were significantly different from those of other samples, which might be attributed to the different production processes of agarwood. For further investigation, factor analysis was conducted for six agarwood samples. The results showed that the data classification possessed a regional characteristic; according to the retention time and relative content, characteristic compositions were determined by factor scores. Finally, the differences of characteristic compositions were simply analyzed, and the reasons were speculated.

Testing Flight-like Pyrolysis Gas Chromatography-Mass Spectrometry as Performed by the Mars Organic Molecule Analyzer Onboard the ExoMars 2020 Rover on Oxia Planum Analog Samples.

The Mars Organic Molecule Analyzer (MOMA) onboard the ExoMars 2020 rover (to be landed in March 2021) utilizes pyrolysis gas chromatography-mass spectrometry (GC-MS) with the aim to detect organic molecules in martian (sub-) surface materials. Pyrolysis, however, may thermally destroy and transform organic matter depending on the temperature and nature of the molecules, thus altering the original molecular signatures. In this study, we tested MOMA flight-like pyrolysis GC-MS without the addition of perchlorates on well-characterized natural mineralogical analog samples for Oxia Planum, the designated ExoMars 2020 landing site. Experiments were performed on an iron-rich shale (that is rich in Fe-Mg-smectites) and an opaline chert, with known organic matter compositions, to test pyrolytic effects related to heating in the MOMA oven. Two hydrocarbon standards (n-octadecane and phytane) were also analyzed. The experiments show that during stepwise pyrolysis (300°C, 500°C, and 700°C), (1) low-molecular-weight hydrocarbon biomarkers (such as acyclic isoprenoids and aryl isoprenoids) can be analyzed intact, (2) discrimination between free and complex molecules (macromolecules) is principally possible, (3) secondary pyrolysis products and carryover may affect the 500°C and 700°C runs, and (4) the type of the organic matter (functionalized vs. defunctionalized) governs the pyrolysis outcome rather than the difference in mineralogy. Although pyrosynthesis reactions and carryover clearly have to be considered in data interpretation, our results demonstrate that pyrolysis GC-MS onboard MOMA operated under favorable conditions (e.g., no perchlorates) will be capable of providing important structural information on organic matter found on Mars, particularly when used in conjunction with other techniques on MOMA, including derivatization and thermochemolysis GC-MS and laser desorption/ionization-MS.

Development of direct microwave desorption/gas chromatography mass spectrometry system for rapid analysis of volatile components in medicinal plants.

A new direct microwave desorption-gas chromatography-mass spectrometry method was developed for the analysis of the essential oils of medicinal plants. A homemade direct microwave desorption system was fabricated and used for the desorption of volatile components of medicinal herbs. The desorbed volatiles are transferred directly into the gas chromatography injector for analysis in a one-step process. Approximately 0.3 g of the herb was needed for the desorption of samples in 60 s. In this study, more than 53 volatile compounds were identified and quantified for Echinophora platyloba DC as model herb sample. The results were found to be in good agreement with the conventional hydrodistillation extraction data. The described results show that direct microwave desorption is fast, simple, and easy to automate and requires only a small amount of sample. The results indicate that essential oil components valuable for varietal identification and characteristic of each variety analyzed when direct microwave desorption-gas chromatography-mass spectrometry was used for analysis.

The geometrical characteristics of nickel-based metal organic framework on its entrapment capability.

Here, a three dimensional nickel-based metal organic framework (MOF) was synthesized via solvothermal and room temperature protocols. In order to study the effects of the synthesis conditions on the physical properties such as pore sizes and shapes of the prepared MOFs, their extraction capabilities were examined. Both MOFs were characterized by Fourier transform infrared spectroscopy, powder X-ray diffraction, scanning electron microscopy, Brunauer-Emmett-Teller and thermogravimetric analyses. Brilliant properties such as porous structure, high surface area and considerable thermal stability make them reasonable candidates to be employed as efficient extractive phases. The efficiency of the superior nickel-based MOF was evaluated for headspace needle trap extraction of chlorobenzenes as model compounds in conjunction with gas chromatography-mass spectrometry (GC-MS). The MOF-based extractive phase was conveniently packed in a needle trap device and after extraction, the desorption process was performed via direct insertion of needle into the GC inlet. After optimizing the extraction/desorption conditions, the figures of merit such as linear dynamic range was in the range of 5-1000 ng L-1 (R2 > 0.987) while the limits of detection and quantification values were 2-10 and 6-30 ng L-1, respectively. The intra- and inter-day relative standard deviations for three replicates at the concentration level of 50 ng L-1 were in the range of 7-9% and 9-12%, respectively. The needle-to-needle reproducibility was also found to be in the range of 5-11%. Acceptable relative recovery values at the concentration level of 50 ng L-1 ranged from 85 to 96%, showing no significant matrix effect.

Sensitivity and resolution optimization in gas chromatography coupled to mass spectrometry analyses of volatile organic compounds present in vacuum environment.

The gaseous phase analyses of volatile organic compounds (VOCs) are an important challenge especially when these organics are formed in high vacuum environments (10-8 mbar) reproducing the environment of astrophysical ices formation and processing. Several analytical techniques have been developed to identify the molecular diversity formed from the processing of these ices. Among them, the coupling of a GC-MS to the vacuum chamber where ices are processed highlighted the interesting chemical diversity of such processed ices. These analyses were possible due to the development of a specific system, the VAHIIA interface that enables the preconcentration of VOCs at low pressure (10-8 mbar) and their transfer at higher pressure to the injection unit of a GC for their subsequent analyses. This system showed sufficient repeatability (13%) and low detection limits (nmol) for simple ices [1], but presents limits when ice mixtures are complex (such as multi-component ices including water, methanol and ammonia). In this contribution, we present the optimization of our previous VAHIIA system by implementing a cryofocusing system in the GC oven and by improving the recovery yield of VOCs from the vacuum chamber to the VAHIIA interface. The cryofocusing provides an improvement of efficiencies leading to higher resolution and signal to noise ratio, while the addition of argon in the vacuum chamber during the VOC recovery allows increasing the amount of molecules recovered by a factor of ∼200. The coupling of both approaches provides an increase of sensitivity of a factor ∼400. At the end, experiments on astrophysical ices are shown demonstrating the interest of such optimizations for VOC analyses.

Lipid-rich fraction of the sclerotium of Tiger Milk Mushroom Lignosus rhinocerotis (Agaricomycetes) attenuates LPS-induced inflammation in BV2 cells via Nrf2 pathway

Lignosus rhinocerotis (tiger milk mushroom) is widely used by the indigenous people of Malaysia as a traditional remedy. The present study was carried out in order to evaluate the antioxidant, cytotoxic and anti-neuroinflammatory activities of L. rhinocerotis extract on brain microglial cells (BV2). The antioxidant activity was evaluated by 2,2-diphenyl-1-picryhydrazyl (DPPH•), 2,2'-azinobis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS•+) scavenging assays, and ferric reducing antioxidant power (FRAP). The FRAP, DPPH and ABTS•+ scavenging capacities of the TE3 fraction were 420.77 mg FE/g, 58.01%, and 7%, respectively. The cytotoxic activity was determined by MTS assay. The in vitro model of anti-neuroinflammatory property was evaluated by measuring the production of nitric oxide (NO) in lipopolysaccharide (LPS)-induced BV2 cells. The TE3 fraction showed a significant NO reduction at 1 to 100 µg/mL. The TE3 fraction down-regulated inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX2) genes while it upregulated heme oxygenase (HO-1) and NADPH quinone acceptor oxidoreductase-1 (NQO-1) genes. The nuclear factor (erythroid-derived 2)-like 2 (Nrf2) transcription was also activated. The chemical component of the active fraction (TE3) was identified by gas chromatography-mass spectrometry (GCMS). Overall, the BV2 in vitro model anti-neuroinflammatory activity of L. rhinocerotis may be caused by the lipid constituents identified in the fraction