Tags

Type your tag names separated by a space and hit enter

Simultaneous determination of 3-nitrotyrosine, tyrosine, hydroxyproline and proline in exhaled breath condensate by hydrophilic interaction liquid chromatography/electrospray ionization tandem mass spectrometry.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Dec 01; 860(1):78-85.JC

Abstract

The analysis of biomarkers from exhaled breath condensate (EBC) is a non-invasive but challenging method for the detection of pulmonary diseases. The amino acids L-proline (Pro) and l-tyrosine (Tyr) are precursors for two important metabolites, trans-L-4-hydroxyproline (trans-L-4-hydroxypyrrolidin-2-carboxylic acid, t-Hyp) and nitrotyrosine (NT). Whereas t-Hyp is supposed to be a biomarker for lung fibrosis, NT is a promising biomarker for inflammation in airway diseases. Analysis of EBC requires extremely sensitive methods, because the epithelial lining fluid of the lung and upper airway is highly diluted in EBC. The high intra- and interindividual variation of this dilution implicates additional problems for sample collection and the interpretation of EBC results. Hence, our aim was to work out a method that would compensate for these possible dilution effects. We have developed a new, reliable and very sensitive method for the simultaneous determination of Pro, t-Hyp, Tyr and NT from EBC. Except for t-Hyp, we used labelled internal standards (IS) L-proline (13)C(5), (15)N (Pro (13)C(5)), L-tyrosine-(13)C(9) (Tyr (13)C(9)), (13)C(9)-3-nitrotyrosine (NT(13)C(9)), IS for t-Hyp was cis-4-hydroxy-L-proline, which were added to the samples before they were lyophilised for concentration. For the separation of the analytes we used hydrophilic interaction liquid chromatography (HILIC), coupled to tandem-mass-spectrometry (MS/MS). The limit of detection (LOD) was 0.5 microg/l for Pro and Tyr and 5 ng/l for t-Hyp and NT. The relative standard deviation (RSD) of the precision from day to day was between 2.6 and 8.0% at spiked concentrations between 4 and 25 microg/l for Pro and between 4.2 and 7.3% for Tyr. The RSD of the precision from day to day was between 7.5 and 13.2% at spiked concentrations between 40 and 250 ng/l for t-Hyp and between 3.5 and 8.2% for NT. The method was established using 27 healthy subjects with a median age of 46 years. Concentrations ranged from 2.8 to 51.9 microg/l for Pro, from <5 to 516.5 ng/l for t-Hyp, from 2.4 to 99.1 for Tyr and for NT concentration ranged between <5 and 1686.5 ng/l.

Authors+Show Affiliations

Institute and Outpatient Clinic of Occupational and Social Medicine, University Hospital, Aachen University of Technology, Pauwelsstrasse 30, D-52074 Aachen, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Validation Study

Language

eng

PubMed ID

18024246

Citation

Conventz, A, et al. "Simultaneous Determination of 3-nitrotyrosine, Tyrosine, Hydroxyproline and Proline in Exhaled Breath Condensate By Hydrophilic Interaction Liquid Chromatography/electrospray Ionization Tandem Mass Spectrometry." Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, vol. 860, no. 1, 2007, pp. 78-85.
Conventz A, Musiol A, Brodowsky C, et al. Simultaneous determination of 3-nitrotyrosine, tyrosine, hydroxyproline and proline in exhaled breath condensate by hydrophilic interaction liquid chromatography/electrospray ionization tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2007;860(1):78-85.
Conventz, A., Musiol, A., Brodowsky, C., Müller-Lux, A., Dewes, P., Kraus, T., & Schettgen, T. (2007). Simultaneous determination of 3-nitrotyrosine, tyrosine, hydroxyproline and proline in exhaled breath condensate by hydrophilic interaction liquid chromatography/electrospray ionization tandem mass spectrometry. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 860(1), 78-85.
Conventz A, et al. Simultaneous Determination of 3-nitrotyrosine, Tyrosine, Hydroxyproline and Proline in Exhaled Breath Condensate By Hydrophilic Interaction Liquid Chromatography/electrospray Ionization Tandem Mass Spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Dec 1;860(1):78-85. PubMed PMID: 18024246.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simultaneous determination of 3-nitrotyrosine, tyrosine, hydroxyproline and proline in exhaled breath condensate by hydrophilic interaction liquid chromatography/electrospray ionization tandem mass spectrometry. AU - Conventz,A, AU - Musiol,A, AU - Brodowsky,C, AU - Müller-Lux,A, AU - Dewes,P, AU - Kraus,T, AU - Schettgen,T, Y1 - 2007/11/19/ PY - 2007/08/14/received PY - 2007/10/08/revised PY - 2007/10/10/accepted PY - 2007/11/21/pubmed PY - 2008/3/5/medline PY - 2007/11/21/entrez SP - 78 EP - 85 JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences JO - J Chromatogr B Analyt Technol Biomed Life Sci VL - 860 IS - 1 N2 - The analysis of biomarkers from exhaled breath condensate (EBC) is a non-invasive but challenging method for the detection of pulmonary diseases. The amino acids L-proline (Pro) and l-tyrosine (Tyr) are precursors for two important metabolites, trans-L-4-hydroxyproline (trans-L-4-hydroxypyrrolidin-2-carboxylic acid, t-Hyp) and nitrotyrosine (NT). Whereas t-Hyp is supposed to be a biomarker for lung fibrosis, NT is a promising biomarker for inflammation in airway diseases. Analysis of EBC requires extremely sensitive methods, because the epithelial lining fluid of the lung and upper airway is highly diluted in EBC. The high intra- and interindividual variation of this dilution implicates additional problems for sample collection and the interpretation of EBC results. Hence, our aim was to work out a method that would compensate for these possible dilution effects. We have developed a new, reliable and very sensitive method for the simultaneous determination of Pro, t-Hyp, Tyr and NT from EBC. Except for t-Hyp, we used labelled internal standards (IS) L-proline (13)C(5), (15)N (Pro (13)C(5)), L-tyrosine-(13)C(9) (Tyr (13)C(9)), (13)C(9)-3-nitrotyrosine (NT(13)C(9)), IS for t-Hyp was cis-4-hydroxy-L-proline, which were added to the samples before they were lyophilised for concentration. For the separation of the analytes we used hydrophilic interaction liquid chromatography (HILIC), coupled to tandem-mass-spectrometry (MS/MS). The limit of detection (LOD) was 0.5 microg/l for Pro and Tyr and 5 ng/l for t-Hyp and NT. The relative standard deviation (RSD) of the precision from day to day was between 2.6 and 8.0% at spiked concentrations between 4 and 25 microg/l for Pro and between 4.2 and 7.3% for Tyr. The RSD of the precision from day to day was between 7.5 and 13.2% at spiked concentrations between 40 and 250 ng/l for t-Hyp and between 3.5 and 8.2% for NT. The method was established using 27 healthy subjects with a median age of 46 years. Concentrations ranged from 2.8 to 51.9 microg/l for Pro, from <5 to 516.5 ng/l for t-Hyp, from 2.4 to 99.1 for Tyr and for NT concentration ranged between <5 and 1686.5 ng/l. SN - 1570-0232 UR - https://www.unboundmedicine.com/medline/citation/18024246/Simultaneous_determination_of_3_nitrotyrosine_tyrosine_hydroxyproline_and_proline_in_exhaled_breath_condensate_by_hydrophilic_interaction_liquid_chromatography/electrospray_ionization_tandem_mass_spectrometry_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1570-0232(07)00707-6 DB - PRIME DP - Unbound Medicine ER -