Gauging circadian variation in ketamine metabolism by real-time breath analysis

The time-of-day of drug application is an important factor in maximizing efficacy and minimizing toxicity. Real-time in vivo mass spectrometric breath analysis of mice was deployed to investigate time-of-day variation in ketamine metabolism. Different production rates of ketamine metabolites, including the recently described anti-depressant hydroxynorketamine, were found in opposite circadian phases. Thus, breath analysis has potential as a rapid and 3Rs (Replacement, Reduction and Refinement) conforming screening method to estimate the time-dependence of drug metabolism

A gas-phase standard delivery system for direct breath analysis

Applications for direct breath analysis by mass spectrometry (MS) are rapidly expanding. One of the more recent mass spectrometry-based approaches is secondary electrospray ionization coupled to high-resolution mass spectrometry (SESI-HRMS). Despite increasing usage, the SESI methodology still lacks standardization procedures for quality control and absolute quantification. In this study, we designed and evaluated a custom-built standard delivery system tailored for direct breath analysis. The system enables the simultaneous introduction of multiple gas-phase standard compounds into ambient MS setups in the lower parts-per-million (ppm) to parts-per-billion (ppb) range. To best mimic exhaled breath, the gas flow can be heated (37 °C–40 °C) and humidified (up to 98% relative humidity). Inter-laboratory comparison of the system included various SESI-HRMS setups, i.e. an Orbitrap and a quadrupole time-of-flight mass spectrometer (QTOF), and using both single- as well as multi-component standards. This revealed highly stable and reproducible performances with between-run variation 95%) for the single-compound standard acetone, while compound-specific performances were obtained for the multi-component standard. Similarly, the sensitivity varied for different compounds within the multi-component standard across all SESI-Orbitrap and -QTOF setups, yielding limits of detections from 3.1 ppb (for p-xylene) to 0.05 ppb (for 1,8-cineol). Routinely applying the standard system throughout several weeks, allowed us to monitor instrument stability and to identify technical outliers in exhaled breath measurements. Such routine deployment of standards would significantly improve data quality and comparability, which is especially important in longitudinal and multi-center studies. Furthermore, performance validation of the system demonstrated its suitability for reliable absolute quantification while it illustrated compound-dependent behavior for SESI

Identification of Exhaled Metabolites in Children with Cystic Fibrosis

The early detection of inflammation and infection is important to prevent irreversible lung damage in cystic fibrosis. Novel and non-invasive monitoring tools would be of high benefit for the quality of life of patients. Our group previously detected over 100 exhaled mass-to-charge (m/z) features, using on-line secondary electrospray ionization high-resolution mass spectrometry (SESI-HRMS), which distinguish children with cystic fibrosis from healthy controls. The aim of this study was to annotate as many m/z features as possible with putative chemical structures. Compound identification was performed by applying a rigorous workflow, which included the analysis of on-line MS2 spectra and a literature comparison. A total of 49 discriminatory exhaled compounds were putatively identified. A group of compounds including glycolic acid, glyceric acid and xanthine were elevated in the cystic fibrosis group. A large group of acylcarnitines and aldehydes were found to be decreased in cystic fibrosis. The proposed compound identification workflow was used to identify signatures of volatile organic compounds that discriminate children with cystic fibrosis from healthy controls, which is the first step for future non-invasive and personalized applications. Keywords: SESI-HRMS; breath analysis; children; cystic fibrosis; infection; inflammation; putative compound identificatio

Status of selected ion flow tube MS: accomplishments and challenges in breath analysis and other areas

This article reflects our observations of recent accomplishments made using selected ion flow tube MS (SIFT-MS). Only brief descriptions are given of SIFT-MS as an analytical method and of the recent extensions to the underpinning analytical ion chemistry required to realize more robust analyses. The challenge of breath analysis is given special attention because, when achieved, it renders analysis of other air media relatively straightforward. Brief overviews are given of recent SIFT-MS breath analyses by leading research groups, noting the desirability of detection and quantification of single volatile biomarkers rather than reliance on statistical analyses, if breath analysis is to be accepted into clinical practice. A 'strengths, weaknesses, opportunities and threats' analysis of SIFT-MS is made, which should help to increase its utility for trace gas analysis

An interoperability framework for multicentric breath metabolomic studies

Exhaled breath contains valuable information at the molecular level and offers promising potential for precision medicine. However, few breath tests transition to routine clinical practice, partly because of the missing validation in multicenter trials. Therefore, we developed and applied an interoperability framework for standardized multicenter data acquisition and processing for breath analysis with secondary electrospray ionization-high resolution mass spectrometry. We aimed to determine the technical variability and metabolic coverage. Comparison of multicenter data revealed a technical variability of ∼20% and a core signature of the human exhaled metabolome consisting of ∼850 features, corresponding mainly to amino acid, xenobiotic, and carbohydrate metabolic pathways. In addition, we found high inter-subject variability for certain metabolic classes (e.g., amino acids and fatty acids), whereas other regions such as the TCA cycle were relatively stable across subjects. The interoperability framework and overview of metabolic coverage presented here will pave the way for future large-scale multicenter trials

Real-time monitoring of exhaled volatiles using atmospheric pressure chemical ionization on a compact mass spectrometer

© 2016 Future Science Ltd.Aim: Breath analyses have potential to detect early signs of disease onset. Ambient ionization allows direct combination of breath gases with MS for fast, on-line analysis. Portable MS systems would facilitate field/clinic-based breath analyses. Results & methodology: Volunteers ingested peppermint oil capsules and exhaled volatile compounds were monitored over 10 h using a compact mass spectrometer. A rise and fall in exhaled menthone was observed, peaking at 60-120 min. Real-time analysis showed a gradual rise in exhaled menthone postingestion. Sensitivity was comparable to established methods, with detection in the parts per trillion range. Conclusion: Breath volatiles were readily analyzed on a portable mass spectrometer through a simple inlet modification. Induced changes in exhaled profiles were detectable with high sensitivity and measurable in real-time

SIFT-MS and FA-MS methods for ambient gas phase analysis: developments and applications in the UK

Selected ion flow tube mass spectrometry, SIFT-MS, a relatively new gas/vapour phase analytical method, is derived from the much earlier selected ion flow tube, SIFT, used for the study of gas phase ion-molecule reactions. Both the SIFT and SIFT-MS techniques were conceived and developed in the UK, the former at Birmingham University, the latter at Keele University along with the complementary flowing afterglow mass spectrometry, FA-MS, technique. The focus of this short review is largely to describe the origins, developments and, most importantly, the unique features of SIFT-MS as an analytical tool for ambient analysis and to indicate its growing use to analyse humid air, especially exhaled breath, its unique place as a on-line, real time analytical method and its growing use and applications as a non-invasive diagnostic in clinical diagnosis and therapeutic monitoring, principally within several UK universities and hospitals, and briefly in the wider world. A few case studies are outlined that show the potential of SIFT-MS and FA-MS in the detection and quantification of metabolites in exhaled breath as a step towards recognising pathophysiology indicative of disease and the presence of bacterial and fungal infection of the airways and lungs. Particular cases include the detection of Pseudomonas aeruginosa infection of the airways of patients with cystic fibrosis (SIFT-MS) and the measurement of total body water in patients with chronic kidney disease (FA-MS). The growing exploitation of SIFT-MS in other areas of research and commerce are briefly listed to show the wide utility of this unique UK-developed analytical method, and future prospects and developments are alluded to

The peppermint breath test benchmark for PTR-MS and SIFT-MS

18openInternationalInternational coauthor/editorA major challenge for breath research is the lack of standardization in sampling and analysis. To address this, a test that utilizes a standardized intervention and a defined study protocol has been proposed to explore disparities in breath research across different analytical platforms and to provide benchmark values for comparison. Specifically, the Peppermint Experiment involves the targeted analysis in exhaled breath of volatile constituents of peppermint oil after ingestion of the encapsulated oil. Data from the Peppermint Experiment performed by proton transfer reaction mass spectrometry (PTR-MS) and selected ion flow tube mass spectrometry (SIFT-MS) are presented and discussed herein, including the product ions associated with the key peppermint volatiles, namely limonene, α- and β-pinene, 1,8-cineole, menthol, menthone and menthofuran. The breath washout profiles of these compounds from 65 individuals were collected, comprising datasets from five PTR-MS and two SIFT-MS instruments. The washout profiles of these volatiles were evaluated by comparing the log-fold change over time of the product ion intensities associated with each volatile. Benchmark values were calculated from the lower 95% confidence interval of the linear time-to-washout regression analysis for all datasets combined. Benchmark washout values from PTR-MS analysis were 353 min for the sum of monoterpenes and 1,8-cineole (identical product ions), 173 min for menthol, 330 min for menthofuran, and 218 min for menthone; from SIFT-MS analysis values were 228 min for the sum of monoterpenes, 281 min for the sum of monoterpenes and 1,8-cineole, and 370 min for menthone plus 1,8-cineole. Large inter- and intra-dataset variations were observed, whereby the latter suggests that biological variability plays a key role in how the compounds are absorbed, metabolized and excreted from the body via breath. This variability seems large compared to the influence of sampling and analytical procedures, but further investigations are recommended to clarify the effects of these factors.openHenderson, Ben; Slingers, Gitte; Pedrotti, Michele; Pugliese, Giovanni; Malásková, Michaela; Bryant, Luke; Lomonaco, Tommaso; Ghimenti, Silvia; Moreno, Sergi; Cordell, Rebecca; Harren, Frans J M; Schubert, Jochen; Mayhew, Chris A; Wilde, Michael; Di Francesco, Fabio; Koppen, Gudrun; Beauchamp, Jonathan D; Cristescu, Simona MHenderson, B.; Slingers, G.; Pedrotti, M.; Pugliese, G.; Malásková, M.; Bryant, L.; Lomonaco, T.; Ghimenti, S.; Moreno, S.; Cordell, R.; Harren, F.J.M.; Schubert, J.; Mayhew, C.A.; Wilde, M.; Di Francesco, F.; Koppen, G.; Beauchamp, J.D.; Cristescu, S.M