DOI: 10.5281/zenodo.6476097
title: Exhaled breath condensate samples (mzML files) analysed by uHPLC-ESI-OrbitrapMS

description: LC-MS raw data files<strong> </strong>were converted to mzML using MSConvert (Version: 3.0.20279, ProteoWizard) - QC_pos_xxx: QC samples analysed in positive polarity (11 files)<br> - ACOS_XXX_pos_01 to 03: Asthmatic patients (3 replicates) randomly injected (15 files)<br> - Ctrl_XXX_pos_01 to 03: Control (15 files)<br> - DPOC_XXX_pos_01 to 03: COPD (15 files) <strong>EBC Samples and Clinical Assessment</strong> EBC samples were collected from 15 individuals randomly selected (five controls, five with asthma medical diagnosis, and five with COPD medical diagnosis, as assessed by the OLDER Study – Obstructive Lung Diseases in Elders). The Ethics Committee of Nova Medical School approved this study. Asthma was assigned when the patient reported respiratory symptoms, was a nonsmoker, and presented a positive reversibility test (FEV1 &gt; 12% and 200 mL). COPD disease was attributed to those who also reported being current smokers, had a post-bronchodilator FEV1/FVC &lt; 0.70, and had a negative reversibility test. <strong>LC–MS Analysis</strong> Samples were analyzed in triplicate by LC–MS using an Orbitrap Q Exactive Focus (Thermo Scientific) coupled to an Ultimate 3000 UHPLC (Thermo Scientific). A pooled quality control (QC) sample was used to compensate for any possible time-dependent batch effects. The QC samples were created using a small aliquot from each sample. The QC samples were reinjected at regular intervals to bracket the samples. The separation was performed using a Waters XBridge column C18 (2.1 × 150 mm, 3.5 μm particle size, P/N 186003023). The mobile phase A was water with 0.1% formic acid (v/v), and mobile phase B was acetonitrile with 0.1% formic acid (v/v) (Optima LC–MS Grade, Fisher Scientific). The gradient program was as follows: 1 min at 1% B; 1–13 min from 1 to 99% B, 13–15 min at 99% B, 15–16 min from 99 to 1% B, and 4 min at 1% B. The column temperature was maintained at 30 °C, and a flow rate of 400 μL/min was used. The Q Exactive Focus MS method consisted of several cycles of full MS scan (<em>R</em> = 70000) followed by three ddMS2 scans (<em>R</em> = 17500), with a (N)CE of 30 and in positive mode. External calibration was performed using LTQ ESI Positive Ion Calibration Solution (Thermo Scientific) and the lock mass enabled internal calibration. Data were obtained using the Xcalibur software v.4.0.27.19 (Thermo Scientific). The raw MS files, as recorded by the instrument, are available from the corresponding author upon reasonable request.
description: If using please cite - ACS Omega 2020, 5, 26, 16089–16098 https://doi.org/10.1021/acsomega.0c01610 Acknowledgements This work was financially supported by: (i) LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE); (ii) projects POCI-01-0145-FEDER-029702 and POCI-01-0145-FEDER-031297 funded by FEDER funds through COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI) and by national funds (PIDDAC) through FCT/MCTES; (iii) AstraZeneca – Projecto OLDER (CEDOC/2015/59); (iv) iNOVA4Health (UIDB/04462/2020, UIDP/04462/2020) and LS4FUTURE Associated Laboratory (LA/P/0087/2020), financially supported by FCT - Fundação para a Ciência e a Tecnologia, I.P.