Rate normalization for sweat metabolomics biomarker discovery.

As the demand for real-time exercise performance feedback increases, excreted sweat has become a biosource of interest for continuous human performance assessment. For sweat to truly fulfill this requirement, analyte concentrations must be normalized to adequately assess day-to-day differences within and among individuals. In this manuscript, data are presented highlighting the use of accurate localized sweat rate as a means for ion and global metabolomic data normalization. The results illustrate large sweat rate variability among individuals over the course of two distinct exercises protocols. Furthermore, the data show sweat rate is not symmetrical at similar locations among right and left forearms of individuals (p = 0.0007). Sweat ion conductivity analysis suggest overall sweat rate normalization reduces variability collectively among ion values and participants with principal component analysis showing 77.8% of variation in the data set attributable to sweat rate normalization. Global metabolomic analysis of sweat illustrated overall rate normalization increases the variability among test subjects with 72.7% of the variation explained by sweat rate normalization. Finally, overall rate normalized metabolomic features of sweat significantly correlated (ρ ≥ 0.7, ρ ≤ -0.7) with measured performance metrics of the individual, establishing the potential for sweat to be used as a biosource for performance monitoring. Collectively, these data illustrate the importance of accurate localized sweat rate determination, for analyte data normalization, in support for the use of sweat in biomarker discovery efforts to predict human performance.

Evaluation of a standardized collection device for exhaled breath sampling onto thermal desorption tubes.

The Respiration Collector for In Vitro Analysis (ReCIVA) sampler, marketed by Owlstone Medical, provides a step forward in exhaled breath sampling through active sampling directly onto thermal desorption (TD) tubes. Although an improvement to the issues surrounding breath bag sampling, the ReCIVA device, first released in 2015, is a relatively new research and clinical tool that requires further exploration. Here, data are presented comparing two distinct ReCIVA devices. The results, comparing ReCIVA serial numbers #33 and #65, demonstrate that overall statistically insignificant results are obtained via targeted isoprene quantitation (p > 0.05). However, when the data are parsed by the TD tube type used to capture breath volatiles, either Tenax TA or the dual bed Tenax/Carbograph 5TD (5TD), a statistical difference (p < 0.05) among the two different TD tubes was present. These data, comparing the two ReCIVA devices with both Tenax TA and 5TD tubes, are further supported by a global metabolomics analysis yielding 85% of z-scores, comparing ReCIVA devices, below the limit for significance. Experiments to determine the effect of breathing rate on ReCIVA function, using guided breathing for low (7.5 breaths min-1) and high (15 breaths min-1) breathing rates, demonstrate the ReCIVA device shows no statistical difference among breathing rates for quantitated isoprene (p > 0.05). Global metabolomics analysis of the guided breathing rate data shows more than 87% of the z-scores, comparing high and low breathing rates using both the Tenax and the 5TD tubes, are below the level for significance. Finally, data are provided from a single participant who displayed background levels of isoprene while illustrating levels of acetone consistent with the remaining participants. Collectively, these data support the use of multiple ReCIVA devices for exhaled breath collection and provide evidence for an instance where exhaled isoprene is consistent with background levels.

Characterization of standardized breath sampling for off-line field use.

Due to several sources of potential variability associated with exhaled breath bag sampling procedures for off-line analysis, the Respiration Collector for in vitro Analysis (ReCIVA) sampler was developed. Although designed to improve upon several pitfalls of sampling with exhaled breath bags, the ReCIVA remains a minimally studied research tool. In this manuscript, several attributes of the ReCIVA sampler are investigated among three individual tests, such as background contamination, control software version, performance of different adsorbent tubes, duplicate sample production, and comparison to exhaled breath bags. The data shows greater than a 58% reduction in background siloxanes can be achieved with submersion of ReCIVA masks in ethyl alcohol or baking the masks at a high temperature (200 °C). The results illustrate the ReCIVA control software version plays a key role in the flow rates applied to thermal desorption (TD) tubes. Using exhaled isoprene as a representative analyte, the data suggest duplicate samples among ReCIVA pump banks can be achieved using two different thermal desorption tubes, Tenax TA and Tenax/Carbograph 5TD, when using an updated control software and manually calibrating the ReCIVA pumps to uniform flow rates (Tenax p = 0.3869, 5TD p = 0.3131). Additionally, using the updated control software and manual ReCIVA flow calibration, the data suggest the ReCIVA can produce statistically similar results among TD tube types (p = 0.3824) and compared to standard exhaled breath bags (p = 0.1534). Collectively, these results establish a method for manually calibrating the flow of the ReCIVA device to allow for the most consistent results. These data support further experimentation into the use of the ReCIVA sampler for exhaled breath research.