Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate
Abstract Background Researchers investigating lung diseases, such as asthma, have questioned whether certain compounds previously reported in exhaled breath condensate (EBC) originate from saliva contamination. Moreover, despite its increasing use in ‘omics profiling studies, the constituents of EBC...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-04-01
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| Series: | Respiratory Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12931-017-0538-5 |
| _version_ | 1818126754012725248 |
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| author | Charmion Cruickshank-Quinn Michael Armstrong Roger Powell Joe Gomez Marc Elie Nichole Reisdorph |
| author_facet | Charmion Cruickshank-Quinn Michael Armstrong Roger Powell Joe Gomez Marc Elie Nichole Reisdorph |
| author_sort | Charmion Cruickshank-Quinn |
| collection | DOAJ |
| description | Abstract Background Researchers investigating lung diseases, such as asthma, have questioned whether certain compounds previously reported in exhaled breath condensate (EBC) originate from saliva contamination. Moreover, despite its increasing use in ‘omics profiling studies, the constituents of EBC remain largely uncharacterized. The present study aims to define the usefulness of EBC in investigating lung disease by comparing EBC, saliva, and saliva-contaminated EBC using targeted and untargeted mass spectrometry and the potential of metabolite loss from adsorption to EBC sample collection tubes. Methods Liquid chromatography mass spectrometry (LC-MS) was used to analyze samples from 133 individuals from three different cohorts. Levels of amino acids and eicosanoids, two classes of molecules previously reported in EBC and saliva, were measured using targeted LC-MS. Cohort 1 was used to examine contamination of EBC by saliva. Samples from Cohort 1 consisted of clean EBC, saliva-contaminated EBC, and clean saliva from 13 healthy volunteers; samples were analyzed using untargeted LC-MS. Cohort 2 was used to compare eicosanoid levels from matched EBC and saliva collected from 107 asthmatic subjects. Samples were analyzed using both targeted and untargeted LC-MS. Cohort 3 samples consisted of clean-EBC collected from 13 subjects, including smokers and non-smokers, and were used to independently confirm findings; samples were analyzed using targeted LC-MS, untargeted LC-MS, and proteomics. In addition to human samples, an in-house developed nebulizing system was used to determine the potential for EBC samples to be contaminated by saliva. Results Out of the 400 metabolites detected in both EBC and saliva, 77 were specific to EBC; however, EBC samples were concentrated 20-fold to achieve this level of sensitivity. Amino acid concentrations ranged from 196 pg/mL – 4 μg/mL (clean EBC), 1.98 ng/mL – 6 μg/mL (saliva-contaminated EBC), and 13.84 ng/mL – 1256 mg/mL (saliva). Eicosanoid concentration ranges were an order of magnitude lower; 10 pg/mL – 76.5 ng/mL (clean EBC), 10 pg/mL – 898 ng/mL (saliva-contaminated EBC), and 2.54 ng/mL – 272.9 mg/mL (saliva). Although the sample size of the replication cohort (Cohort 3) did not allow for statistical comparisons, two proteins and 19 eicosanoids were detected in smoker vs. non-smoker clean-EBC. Conclusions We conclude that metabolites are present and detectable in EBC using LC-MS; however, a large starting volume of sample is required. |
| first_indexed | 2024-12-11T07:06:29Z |
| format | Article |
| id | doaj.art-715be2776543403bb636b9ff101b0623 |
| institution | Directory Open Access Journal |
| issn | 1465-993X |
| language | English |
| last_indexed | 2024-12-11T07:06:29Z |
| publishDate | 2017-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Respiratory Research |
| spelling | doaj.art-715be2776543403bb636b9ff101b06232022-12-22T01:16:29ZengBMCRespiratory Research1465-993X2017-04-0118112210.1186/s12931-017-0538-5Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensateCharmion Cruickshank-Quinn0Michael Armstrong1Roger Powell2Joe Gomez3Marc Elie4Nichole Reisdorph5Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical CampusSkaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical CampusSkaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical CampusSkaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical CampusSkaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical CampusSkaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical CampusAbstract Background Researchers investigating lung diseases, such as asthma, have questioned whether certain compounds previously reported in exhaled breath condensate (EBC) originate from saliva contamination. Moreover, despite its increasing use in ‘omics profiling studies, the constituents of EBC remain largely uncharacterized. The present study aims to define the usefulness of EBC in investigating lung disease by comparing EBC, saliva, and saliva-contaminated EBC using targeted and untargeted mass spectrometry and the potential of metabolite loss from adsorption to EBC sample collection tubes. Methods Liquid chromatography mass spectrometry (LC-MS) was used to analyze samples from 133 individuals from three different cohorts. Levels of amino acids and eicosanoids, two classes of molecules previously reported in EBC and saliva, were measured using targeted LC-MS. Cohort 1 was used to examine contamination of EBC by saliva. Samples from Cohort 1 consisted of clean EBC, saliva-contaminated EBC, and clean saliva from 13 healthy volunteers; samples were analyzed using untargeted LC-MS. Cohort 2 was used to compare eicosanoid levels from matched EBC and saliva collected from 107 asthmatic subjects. Samples were analyzed using both targeted and untargeted LC-MS. Cohort 3 samples consisted of clean-EBC collected from 13 subjects, including smokers and non-smokers, and were used to independently confirm findings; samples were analyzed using targeted LC-MS, untargeted LC-MS, and proteomics. In addition to human samples, an in-house developed nebulizing system was used to determine the potential for EBC samples to be contaminated by saliva. Results Out of the 400 metabolites detected in both EBC and saliva, 77 were specific to EBC; however, EBC samples were concentrated 20-fold to achieve this level of sensitivity. Amino acid concentrations ranged from 196 pg/mL – 4 μg/mL (clean EBC), 1.98 ng/mL – 6 μg/mL (saliva-contaminated EBC), and 13.84 ng/mL – 1256 mg/mL (saliva). Eicosanoid concentration ranges were an order of magnitude lower; 10 pg/mL – 76.5 ng/mL (clean EBC), 10 pg/mL – 898 ng/mL (saliva-contaminated EBC), and 2.54 ng/mL – 272.9 mg/mL (saliva). Although the sample size of the replication cohort (Cohort 3) did not allow for statistical comparisons, two proteins and 19 eicosanoids were detected in smoker vs. non-smoker clean-EBC. Conclusions We conclude that metabolites are present and detectable in EBC using LC-MS; however, a large starting volume of sample is required.http://link.springer.com/article/10.1186/s12931-017-0538-5EBCSalivaLungMetabolomicsProteomicsLC-MS |
| spellingShingle | Charmion Cruickshank-Quinn Michael Armstrong Roger Powell Joe Gomez Marc Elie Nichole Reisdorph Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate Respiratory Research EBC Saliva Lung Metabolomics Proteomics LC-MS |
| title | Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate |
| title_full | Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate |
| title_fullStr | Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate |
| title_full_unstemmed | Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate |
| title_short | Determining the presence of asthma-related molecules and salivary contamination in exhaled breath condensate |
| title_sort | determining the presence of asthma related molecules and salivary contamination in exhaled breath condensate |
| topic | EBC Saliva Lung Metabolomics Proteomics LC-MS |
| url | http://link.springer.com/article/10.1186/s12931-017-0538-5 |
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