This paper presents developments in stationary phase coatings for microelectromechanical system gas chromatography (MEMS GC). Specifically, we present the coating of MEMS GC separation columns with a chiral stationary phase for the separation of amino acid enantiomers. Three commercial columns coated with chiral stationary phases from Restek were tested: Rt-βDEXm, Rt-βDEXsm, and Rt-βDEXsa. Four amino acid enantiomers (d- and l-) were tested with the 3 commercial columns: alanine (Ala), valine (Val), leucine (Leu), and aspartic acid (Asp). The Rt-βDEXsm column provided the best experimental performance with separation of d- and l-Ala and partial separation of d- and l-Asp. The resolution, Rs, values were 4.65 for the Ala enantiomers and 0.98 for the Asp enantiomers, respectively. The Rt-βDEXsm chiral stationary phase was dynamically coated on three 10-m-long microcolumns connected in series to investigate amino acid enantiomer separation. Successful separation of d- and l-Ala and partial separation of d- and l-Asp were observed with the microcolumns. The Rs values from the chiral-stationary-phase-coated microcolumns were 1.21 and 0.553 for the Ala and Asp enantiomers, respectively. The chromatographically separated amino acid enantiomers were detected by the MAss Spectrometer for Planetary EXploration (MASPEX), a spaceflight mass spectrometer. Future work is required for improving the MEMS GC separation column performance consisting of testing static versus dynamic coating methods and more rigorous investigation of the stationary phase coating thickness. A discussion is provided on future work for the development of an MEMS GC suite targeting broad analyte selectivity for future space science missions.

We present a novel, innovative approach to gas chromatography-mass spectrometry (GC-MS) based on micro-electro-mechanical systems (MEMS) columns that improve the current, state-of-the-art by dramatically reducing the size, mass, and power resources for deploying GC for future landed missions. The outlet of the MEMS GC column was coupled to a prototype of the MAss Spectrometer for Planetary EXploration (MASPEX) through a heated transfer line into the ion source. MEMS GC-MS experiments were performed to demonstrate linearity of response and establish limit of detection (LOD) to alkanes (organics), fatty acid methyl esters (FAMEs) and chemically derivatized amino acids (biological molecules). Linearity of response to each chemical family was demonstrated over two orders of magnitude dynamic range and limit of detection (LOD) values were single to tens (4–43) of picomoles per 1 μl injection volume. MEMS GC column analytical performance was also demonstrated for a “Mega Mix” of chemical analytes including organics and biological molecules. Chromatographic resolution exceeded 200, retention time reproducibility was << 1% RSD (majority ≤ 0.3%), and peak capacity values calculated to be 124 ± 2 over a 435 s retention time window. The 5.5 m MEMS column was also shown to be a suitable alternative to traditional commercial columns for use in comprehensive two-dimensional gas chromatography (GC × GC). Mass spectra collected from MASPEX showed close consistency with National Institute of Technology (NIST) reference mass spectra and were used for high confidence identification of all eluting analytes.

3D3) Measurement of simple molecules with multiple rare ("clumped") isotopes

The mass spectrometer for planetary exploration (MASPEX) is a versatile mass spectrometer with unprecedented mass resolution designed for spaceflight. However, the current version of MASPEX is designed for continuous sampling during flybys of planetary bodies and does not include gas chromatography, which can improve the analysis of complex mixtures of organic compounds in space environments. Here, micro-electro-mechanical system (MEMS) gas chromatography (GC) linearity, reproducibility, and column analytical performance were first demonstrated prior to the coupling to MASPEX for MEMS GC-mass spectrometer (MS) analyses. Linearity of response was demonstrated for n-hexane over 2 orders of magnitude of on-column mass (concentration). Retention time reproducibility in the MEMS GC was <= 2% relative standard deviation (RSD). MEMS GC column analytical performance calculations showed the average number of theoretical plates, N, and the height equivalent to a theoretical plate, HETP, to be 16 239 (1623 plates per meter) and 0.062 cm, respectively. N defines a chromatographic centroid peak apex divided by the peak width at half height, similar to mass resolution. When coupled to MASPEX, the retention time reproducibility was in a similar range of 1-2% relative standard deviation with a slightly larger deviation seen from the mass spectrometer detector due to start trigger variations with a manual start trigger in the FastFlight software compared to the LabView code used for the MEMS GC-MS testing. The collected mass spectra showed close consistency with National Institute of Standards and Technology (NIST) reference mass spectra providing confidence in chemical compound identification. We present the first data generated from the coupling of these devices.

One of the most fundamental measurements that an atmospheric probe can make is the noble gas composition of the well-mixed atmosphere. The relatively inert chemical nature of noble gases makes them excellent tracers of the original reservoirs of material that formed the planet. Three types of measurements are important in this context: 1) abundance relative to the background H2 atmosphere; 2) relative abundances of He, Ne, Ar, Kr, and Xe; and 3) isotopic abundance patterns within individual noble gases. In the latter context, comparison of the Xe isotopic abundances with those from comet 67P by ROSINA/Rosetta provides constraints on the origin of Earth’s volatiles (Marty et al., 2017). The Galileo Probe mass spectrometer made measurements of the noble gases at Jupiter that indicated a factor of 2-3 increase relative to protosolar values. Likewise the noble gas enrichment system, a subsystem of the Galileo Probe mass spectrometer that is based on a SAES-171 getter, provided measurements of He, Ne, and Ar, but did not provide adequate signal-to-noise measurements for Kr and Xe. This task fell to the hydrocarbon enrichment system that used a carbon sieve trapping system (Mahaffy et al., 2000). The criticality of these measurements to the atmospheric probe mission makes it imperative to understand potential issues involved in the Galileo Probe measurements, and indicates the need to use this information to design a more robust gas enrichment system for future probes. Another relevant point concerns the recent observations of ammonia from Juno (Bolton et al., 2017), which suggest that the phase transitions of condensable volatiles as a function of pressure are highly complex at Jupiter and latitude-dependent as a result of dynamical effects. Therefore, of equal importance when considering the sampling systems for the probe mass spectrometer is the need to measure both gaseous and aerosol phases of important condensable compounds such as CH4, NH3, H2S, NH4SH, and H2O that will allow an adequate characterization of the bulk elemental abundances as a function of pressure/altitude. The effects of condensation will be magnified at Saturn and the ice giants owing to greater volatile enrichments and lower atmospheric temperatures. Based on the preceding considerations, the measurement types listed above should have increasing robustness against possible observational artifacts.

Rodent and nonhuman primate studies indicate that developmental programming by reduced perinatal nutrition negatively impacts life course cardio-metabolic health. We have developed a baboon model in which we feed control mothers (CON) ad libitum while nutrient restricted mothers are fed 70% of ad libitum global feed in pregnancy and lactation. Offspring of nutrient restricted mothers are intrauterine growth restricted (IUGR) at term. By 3.5 years IUGR baboons showed signs of insulin resistance, indicating a pre-diabetic phenotype, in contrast to healthy CON offspring. We hypothesized that a novel breath analysis approach would provide markers of the altered cardio-metabolic state in a non-invasive manner. Here we assess whether exhaled breath volatile organic compounds (VOCs) collected from this unique cohort of juvenile baboons with documented cardio-metabolic dysfunction resulting from in utero programming can be detected from their breath signatures. Breath was collected from male and female CON and IUGR baboons at 4.8 ± 0.2 years (human equivalent ∼13 years). Breath VOCs were quantified using a two-dimensional gas chromatography mass spectrometer. Two-way ANOVA, on 76 biologically relevant VOCs identified 27 VOCs (p < 0.05) with altered abundances between groups (sex, birthweight, and sex x birthweight). The 27 VOCs included 2-pentanone, 2-octanone, 2,2,7,7-tetramethyloctane and 3-methyl-1-heptene, which have not previously been associated with cardio-metabolic disease. Unsupervised principal component analysis of these VOCs could discriminate the four clusters defining males, females, CON and IUGR. This study, which is the first to assess quantifiable breath signatures associated with cardio-metabolic programing for any model of IUGR, demonstrates the translational value of this unique model to identify metabolites of programmed cardio-metabolic dysfunction in breath signatures. Future studies are required to validate the translatability of these findings to humans.

Mass spectrometry is a powerful tool to measure the composition of volatile and semi volatile gases. The necessity to accurately identify and quantify unknown species lead to the requirements of a mass spectrometer as the detector of choice in most separation science and direct sample analysis situations. Advantages of time-of-flight mass spectrometry (TOFMS) are the high mass resolution, high mass range, and the measurement of the entire mass range in each extraction. The multi-bounce time-of-flight mass spectrometer (MBTOF) described in this work, takes advantage of a small footprint without sacrificing mass resolution. To achieve this, the MBTOF prototype uses a linear flight path with dual lens stacks. Ions are bounced in between the mirrors for a specified duration whereby increasing their flight time and resolution. The number of bounces can tune the resolution of the instrument. To show the minimum capabilities of the instrument and further applications of it, MBTOF was operated in linear mode. The instrument is designed for a multibounce passage of the ion optics and the focal point of the ion optics is optimized for this application, therefore the resolution in linear mode is limited. However, even in linear mode of operation, the mass resolution meets or exceeds that of a quadrupole mass spectrometer with limited power supplies required for operations. The measurements presented here are based on lab measurements of the early lab prototype MBTOF operated in a linear flight mode with low ion source extraction fields. A detailed evaluation including filament characterization, dynamic range and resolution are investigated. Further discussion involving applications on planetary missions for rocket science, coupling of MBTOF with laser thermal desorption or gas chromatography for potential organic determination in deep space are included.

There is widespread interest in the development of tools to estimate radiation exposures. Exhaled breath provides a novel matrix for assessing biomarkers that could be correlated with exposures. The use of exhaled breath for estimating radiation exposure is warranted, as studies have shown that external exposure to ionizing radiation causes oxidative stress that accelerates lipid peroxidation of polyunsaturated fatty acids, liberating alkanes and alkane metabolites that are excreted in the breath as volatile organic compounds (VOCs). As a proof of principle study, small groups (n = 4) of Gottingen minipigs were whole-body irradiated with gamma rays delivered by a Co-60 source at absorbed doses of 0, 0.25, 0.5, 0.75, 1, 1.25, 2, and 4 Gy. Additional groups (n = 4) were treated with lipopolysaccharide (LPS) or granulocyte colony stimulating factor (G-CSF), with and without concurrent Co-60 exposure, at an absorbed dose of 1 Gy. Breath and background air VOC samples were collected on days -3, -2, -1, 0 pre-irradiation, then at 0.25, 24, 48, 72, and 168 h postirradiation. VOCs were analyzed by automated thermal desorption with two-dimensional gas chromatography and time-of-flight mass spectrometry (ATD GCxGC TOF MS). The results show significant changes in 58 breath VOCs post-irradiation, mainly consisting of methylated and other derivatives of alkanes, alkenes, and benzene. Using a multivariate combination of these VOCs, a radiation response function was constructed, which was significantly elevated at 15 min post irradiation and remained elevated throughout the study (to 168 h post irradiation). As a binary test of radiation absorbed doses >= 0.25 Gy, the radiation response function distinguished irradiated animals fromshams (0 Gy) with 83-84% accuracy. A randomly derived radiation response function was robust: When half of the biomarkers were removed, accuracy was 75%. An optimally derived function with two biomarkers was 82% accurate. As a binary test of radiation absorbed doses >= 0.5 Gy, the radiation response function identified irradiated animals with an accuracy of 87% at 15 min post irradiation and 75.5% at 168 h post irradiation. Treatment with LPS and G-CSF did not affect the radiation response function. This proof-of-principle study supports the hypothesis that breath VOCs may be used for estimating radiation exposures. Further studies will be required to validate the sensitivity and specificity of these potential biomarkers.

In this paper, we describe a simple method for sampling, pre-concentrating, and separating volatile and semi-volatile components from two different cave atmospheres. Sampling is performed by capturing a volume of cave atmosphere in a Tedlar bag or Suma canister for sample storage and transport back to the laboratory. Loading a portion of the sample on a multi-bed sorption trap allows for sample pre-concentration prior to separation and detection of components on a comprehensive two-dimensional gas chromatograph (GC×GC). Comparison of two Texas caves reveals the power of comprehensive two-dimensional gas chromatography (GC×GC) for volatile separation and detection, and to our knowledge marks the first use of GC×GC for the analysis of cave atmospheres. Analysis of the results revealed 138 and 146 chromatographic signals over an S/N threshold of 500 and direct comparison of the two samples revealed 50 identical chromatographic signals. This study is a first step toward demonstrating the ability of GC×GC to separate the complex volatiles and semi-volatiles in the cave atmosphere as a fingerprinting tool.

The use of comprehensive two-dimensional gas chromatography (GC×GC) is becoming more popular for complex sample analysis and the optimization of this technique is paramount for accurate qualitative and quantitative reporting. In this work, the effect of detector data acquisition rate (or detector sampling speed) for GC×GC is investigated and summarized. Effects on chromatographic signal such as peak area, height, and width, as well as analytical figures of merit such as resolution and limit of detection, are investigated at four data acquisition rates using a multi-component test mixture. Adjusting the data acquisition rate of the detector has a profound effect on peak areas, heights, and peak widths. In addition, the increase in detected signal and decreased peak widths led to improved chromatographic resolution and limits of detection (LOD) under optimized conditions.

BackgroundComprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GCxGC-TOF MS) has been proposed as a powerful new tool for multidimensional analysis of complex chemical mixtures. We investigated GCxGC-TOF MS as a new method for identifying volatile organic compounds (VOCs) in normal human breath.MethodsSamples of alveolar breath VOCs and ambient room air VOC were collected with a breath collection apparatus (BCA) onto separate sorbent traps from 34 normal healthy volunteers (mean age = 40 yr, SD = 17 yr, male/female = 19/15). VOCs were separated on two serial capillary columns separated by a cryogenic modulator, and detected with TOF MS. The first and second dimension columns were non-polar and polar respectively.ResultsBCA collection combined with GC×GC-TOF MS analysis identified approximately 2000 different VOCs in samples of human breath, many of which have not been previously reported. The 50 VOCs with the highest alveolar gradients (abundance in breath minus abundance in ambient room air) mostly comprised benzene derivatives, acetone, methylated derivatives of alkanes, and isoprene.ConclusionsCollection and analysis of breath VOCs with the BCA-GC×GC-TOF MS system extended the size of the detectable human volatile metabolome, the volatome, by an order of magnitude compared to previous reports employing one-dimensional GC-MS. The size of the human volatome has been under-estimated in the past due to coelution of VOCs in one-dimensional GC analytical systems.

Breath testing could provide a rational tool for radiation biodosimetry because radiation causes distinct stress-producing molecular damage, notably an increased production of reactive oxygen species. The resulting oxidative stress accelerates lipid peroxidation of polyunsaturated fatty acids, liberating alkanes and alkane metabolites that are excreted in the breath as volatile organic compounds (VOCs). Breath tests were performed before and after radiation therapy over five days in 31 subjects receiving daily fractionated doses: 180-400 cGy d(-1) standard radiotherapy (n = 26), or 700-1200 cGy d(-1) high-dose stereotactic body radiotherapy (n = 5). Breath VOCs were assayed using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry. Multiple Monte Carlo simulations identified approximately 50 VOCs as greater-than-chance biomarkers of radiation on all five days of the study. A consistent subset of 15 VOCs was observed at all time points. A radiation response function was built by combining these biomarkers and the resulting dose-effect curve was significantly elevated at all exposures ⩾1.8 Gy. Cross-validated binary algorithms identified radiation exposures ⩾1.8 Gy with 99% accuracy, and ⩾5 Gy with 78% accuracy. In this proof of principal study of breath VOCs, we built a preliminary radiation response function based on 15 VOCs that appears to identify exposure to localized doses of 1.8 Gy and higher. VOC breath testing could provide a new tool for rapid and non-invasive radiation biodosimetry.

The design and performance evaluation of a two-stage resistively-heated thermal modulator that does not use any cryogenic consumables is described. A commercially available piece of stainless-steel wall coated capillary column is used as the modulator. Cooling of the modulator is provided by a two-stage refrigeration unit with an external heat exchanger and a closed-loop recirculating air system. The modulator is resistively heated by sending a current pulse through each stage of the modulator tube. Studies evaluating the modulator trapping efficiency, cooling efficiency, desorption efficiency, and resultant peak shape are presented and discussed. Performance evaluation of this modulator is compared to similar single-stage modulators, both qualitatively and quantitatively, and has exceeded the previous data reported for the single stage versions. GC x GC chromatograms of a C-6-C-20 n-alkane mixture, a petroleum sample, and a 38-component mixture of aliphatic, aromatic, and halogenated volatile organic compounds are provided. This two-stage modulator proves to be a viable alternative to commercially available modulators which require large amounts of cryogenic fluids.

A novel peak alignment algorithm using a distance and spectrum correlation optimization (DISCO) method has been developed for two-dimensional gas chromatography time-of-flight mass spectrometry (GCxGC/TOF-MS)-based metabolomics. This algorithm uses the output of the instrument control software, ChromaTOF, as its input data. It detects and merges multiple peak entries of the same metabolite into one peak entry in each input peak list. After a z-score transformation of metabolite retention times, DISCO selects landmark peaks from all samples based on both two-dimensional retention times and mass spectrum similarity of fragment ions measured by Pearson's correlation coefficient. A local linear fitting method is employed in the original two-dimensional retention time space to correct retention time shifts. A progressive retention time map searching method is used to align metabolite peaks in all samples together based on optimization of the Euclidean distance and mass spectrum similarity. The effectiveness of the DISCO algorithm is demonstrated using data sets acquired under different experiment conditions and a spiked-in experiment.