Peppermint protocol: first results for gas chromatography-ion mobility spectrometry

ThePeppermint Initiativeseeks to inform the standardisation of breath analysis methods. FivePeppermint Experimentswith gas chromatography-ion mobility spectrometry (GC-IMS), operating in the positive mode with a tritium³H 5.68 keV, 370 MBq ionisation source, were undertaken to provide benchmarkPeppermint Washoutdata for this technique, to support its use in breath-testing, analysis, and research. Headspace analysis of a peppermint-oil capsule by GC-IMS with on-column injection (0.5 cm³) identified 12 IMS responsive compounds, of which the four most abundant were: eucalyptol;β-pinene;α-pinene; and limonene. Elevated concentrations of these four compounds were identified in exhaled-breath following ingestion of a peppermint-oil capsule. An unidentified compound attributed as a volatile catabolite of peppermint-oil was also observed. The most intense exhaled peppermint-oil component was eucalyptol, which was selected as a peppermint marker for benchmarking GC-IMS. Twenty-five washout experiments monitored levels of exhaled eucalyptol, by GC-IMS with on-column injection (0.5 cm³), att= 0 min, and then att+ 60,t+ 90,t+ 165,t+ 285 andt+ 360 min from ingestion of a peppermint capsule resulting in 148 peppermint breath analyses. Additionally, thePeppermint Washoutdata was used to evaluate clinical deployments with a further five washout tests run in clinical settings generating an additional 35 breath samples. Regression analysis yielded an average extrapolated time taken for exhaled eucalyptol levels to return to baseline values to be 429 ± 62 min (±95% confidence-interval). The benchmark value was assigned to the lower 95% confidence-interval, 367 min. Further evaluation of the data indicated that the maximum number of volatile organic compounds discernible from a 0.5 cm³breath sample was 69, while the use of an in-line biofilter appeared to reduce this to 34.