Endogenous Levels of Five Fatty Acid Metabolites in Exhaled Breath Condensate to Monitor Asthma by High-Performance Liquid Chromatography: Electrospray Tandem Mass Spectrometry

Malin L. Nording, Jun Yang, Christine M. Hegedus, Christine M. Hegedus, Abhinav Bhushan, Cristina E Davis, Nicholas Kenyon, Bruce D. Hammock, Malin L. Nording

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Airway inflammation characterizing asthma and other airway diseases may be monitored through biomarker analysis of exhaled breath condensate (EBC). In an attempt to discover novel EBC biomarkers, a high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method was used to analyze EBC from ten control non-asthmatics and one asthmatic individual for five fatty acid metabolites: 9,12,13-trihydroxyoctadecenoic acid (9,12,13-TriHOME), 9,10,13-TriHOME, 12,13-dihydroxyoctade-cenoic acid (12,13-DiHOME), 12-hydroxyeicosatetraenoic acid (12-HETE), and 12(13)-epoxyoctadecenoic acid (12(13)-EpOME). The method was shown to be sensitive, with an on-column limit of quatitation (LOQ) in the pg range (corresponding to pM concentrations in EBC), and linear over several orders of magnitude for each analyte in the calibrated range. Analysis of EBC spiked with the five fatty acid metabolites was within 81%–119% with only a few exceptions. Endogenous levels in EBC exhibited intra- and inter-assay precision of 10%–22%, and 12%–36%, respectively. EBC from the healthy subjects contained average analyte levels between 15 and 180 pM with 12-HETE present above the LOQ in only one of the subjects at a concentration of 240 pM. Exposure of the asthmatic subject to allergen led to increased EBC concentrations of 9,12,13-TriHOME, 9,10,13-TriHOME, 12,13-DiHOME, and 12(13)-EpOME when compared to levels in EBC collected prior to allergen exposure (range =40–510 pM). 12,13-DiHOME was significantly increased (Student's t-test, p < 0.05). In conclusion, we have developed a new HPLC-ESI-MS/MS method for the analysis of five fatty acid metabolites in EBC, which are potential biomarkers for asthma monitoring and diagnosis.

Original languageEnglish (US)
Pages (from-to)123-130
Number of pages8
JournalIEEE Sensors Journal
Volume10
Issue number1
DOIs
StatePublished - 2010

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asthma
metabolites
liquid chromatography
High performance liquid chromatography
fatty acids
Metabolites
Fatty acids
Mass spectrometry
condensates
mass spectroscopy
Biomarkers
Acids
Allergens
biomarkers
Assays
acids
Electrospray ionization
Students
Monitoring
students

Keywords

  • Breath analysis
  • eicosanoids
  • exhaled breath condensate (EBC)
  • inflammation
  • liquid chromatography—mass spectrometry

ASJC Scopus subject areas

  • Instrumentation
  • Electrical and Electronic Engineering

Cite this

Endogenous Levels of Five Fatty Acid Metabolites in Exhaled Breath Condensate to Monitor Asthma by High-Performance Liquid Chromatography : Electrospray Tandem Mass Spectrometry. / Nording, Malin L.; Yang, Jun; Hegedus, Christine M.; Hegedus, Christine M.; Bhushan, Abhinav; Davis, Cristina E; Kenyon, Nicholas; Hammock, Bruce D.; Nording, Malin L.

In: IEEE Sensors Journal, Vol. 10, No. 1, 2010, p. 123-130.

Research output: Contribution to journalArticle

Nording, Malin L. ; Yang, Jun ; Hegedus, Christine M. ; Hegedus, Christine M. ; Bhushan, Abhinav ; Davis, Cristina E ; Kenyon, Nicholas ; Hammock, Bruce D. ; Nording, Malin L. / Endogenous Levels of Five Fatty Acid Metabolites in Exhaled Breath Condensate to Monitor Asthma by High-Performance Liquid Chromatography : Electrospray Tandem Mass Spectrometry. In: IEEE Sensors Journal. 2010 ; Vol. 10, No. 1. pp. 123-130.
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abstract = "Airway inflammation characterizing asthma and other airway diseases may be monitored through biomarker analysis of exhaled breath condensate (EBC). In an attempt to discover novel EBC biomarkers, a high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method was used to analyze EBC from ten control non-asthmatics and one asthmatic individual for five fatty acid metabolites: 9,12,13-trihydroxyoctadecenoic acid (9,12,13-TriHOME), 9,10,13-TriHOME, 12,13-dihydroxyoctade-cenoic acid (12,13-DiHOME), 12-hydroxyeicosatetraenoic acid (12-HETE), and 12(13)-epoxyoctadecenoic acid (12(13)-EpOME). The method was shown to be sensitive, with an on-column limit of quatitation (LOQ) in the pg range (corresponding to pM concentrations in EBC), and linear over several orders of magnitude for each analyte in the calibrated range. Analysis of EBC spiked with the five fatty acid metabolites was within 81{\%}–119{\%} with only a few exceptions. Endogenous levels in EBC exhibited intra- and inter-assay precision of 10{\%}–22{\%}, and 12{\%}–36{\%}, respectively. EBC from the healthy subjects contained average analyte levels between 15 and 180 pM with 12-HETE present above the LOQ in only one of the subjects at a concentration of 240 pM. Exposure of the asthmatic subject to allergen led to increased EBC concentrations of 9,12,13-TriHOME, 9,10,13-TriHOME, 12,13-DiHOME, and 12(13)-EpOME when compared to levels in EBC collected prior to allergen exposure (range =40–510 pM). 12,13-DiHOME was significantly increased (Student's t-test, p < 0.05). In conclusion, we have developed a new HPLC-ESI-MS/MS method for the analysis of five fatty acid metabolites in EBC, which are potential biomarkers for asthma monitoring and diagnosis.",
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