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Standardization procedures for real-time breath analysis by secondary electrospray ionization high-resolution mass spectrometry

机译：二次电喷雾电离高分辨率质谱法实时呼气分析的标准化程序

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Despite the attractiveness of breath analysis as a non-invasive means to retrieve relevant metabolic information, its introduction into routine clinical practice remains a challenge. Among all the different analytical techniques available to interrogate exhaled breath, secondary electrospray ionization high-resolution mass spectrometry (SESI-HRMS) offers a number of advantages (e.g., real-time, yet wide, metabolome coverage) that makes it ideal for untargeted and targeted studies. However, so far, SESI-HRMS has relied mostly on lab-built prototypes, making it difficult to standardize breath sampling and subsequent analysis, hence preventing further developments such as multi-center clinical studies. To address this issue, we present here a number of new developments. In particular, we have characterized a new SESI interface featuring real-time readout of critical exhalation parameters such as CO2, exhalation flow rate, and exhaled volume. Four healthy subjects provided breath specimens over a period of 1month to characterize the stability of the SESI-HRMS system. A first assessment of the repeatability of the system using a gas standard revealed a coefficient of variation (CV) of 2.9%. Three classes of aldehydes, namely 4-hydroxy-2-alkenals, 2-alkenals and 4-hydroxy-2,6-alkedienals?hypothesized to be markers of oxidative stress?were chosen as representative metabolites of interest to evaluate the repeatability and reproducibility of this breath analysis analytical platform. Median and interquartile ranges (IQRs) of CVs for CO2, exhalation flow rate, and exhaled volume were 3.2% (1.5%), 3.1% (1.9%), and 5.0% (4.6%), respectively. Despite the high repeatability observed for these parameters, we observed a systematic decay in the signal during repeated measurements for the shorter fatty aldehydes, which eventually reached a steady state after three/four repeated exhalations. In contrast, longer fatty aldehydes showed a steady behavior, independent of the number of repe
机译：尽管呼吸分析的吸引力作为检索相关的代谢信息的非侵入性手段，但其在常规临床实践中的引入仍然是一个挑战。在询问呼出呼气的所有不同的分析技术中，二次电喷雾电离高分辨率质谱（Sesi-HRMS）提供了许多优点（例如，实时，且宽，代谢覆盖率），使其成为未确定的理想有针对性的研究。然而，到目前为止，SESI-HRMS大多依赖于实验室内置原型，使得难以标准化呼吸采样和随后的分析，因此防止了多中心临床研究等进一步的发展。为了解决这个问题，我们在这里展示了许多新的发展。特别是，我们已经表征了一种新的SESI接口，具有实时读数的临界呼气参数，例如CO2，呼气流量和呼出量。四个健康受试者在1个月的时间内提供呼吸标本，以表征SESI-HRMS系统的稳定性。使用气体标准的系统的重复性评估揭示了2.9％的变异系数（CV）。三类醛类，即4-羟基-2-烯烃，2-烯烃和4-羟基-2,6-烷基因α？假设是氧化应激的标志物？被选择为感兴趣的代表代谢物，以评估可重复性和再现性这种呼吸分析分析平台。 CVS的中位数和四分位数（IQRS）用于CO2，呼气流量和呼出量为3.2％（1.5％），分别为3.1％（1.9％）和5.0％（4.6％）。尽管对这些参数观察到的高可重复性，但我们在短脂肪醛的重复测量期间观察到信号中的系统衰减，这最终在三个/四重复呼气后达到稳定状态。相比之下，较长的脂肪醛显示出稳定的行为，与储存人数无关

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