Development of a new breath collection method for analyzing volatile organic compounds from intubated mouse models

A new pre-clinical method for capturing breath samples from intubated mice is presented. This method significantly reduces background levels, allowing more accurate measurements of VOCs originating from the breath ("on-breath") as opposed to background contamination. The method was developed by integrating industry-standard volatile-capturing sorbent tubes with respiratory mechanics measurement equipment (flexiVent^®), resulting in a mouse breath sample that can be transported and analyzed by TD-GC-MS and other central lab technologies. Using the methodology, the discrimination between on-breath VOCs from background compounds provides a cleaner dataset, which can accelerate the validation of VOCs identified from mouse models and their translation to clinical trials. Three metrics were developed to identify on-breath VOCs, with 22 identified using Type 1 (50% of the breath samples exceeding three standard deviations above the mean signal of the system blanks), 34 with Type 2 (P-value ≤ .05 between paired breath and blank samples), and 61 with Type 3 (ROC-AUC value ≥ 0.8 to differentiate between breath and blank samples). The number of compounds seen at elevated levels on mouse breath was quantified and compared to the levels seen on human breath samples to compare methodologies.