Determination of hydrogen peroxide in exhaled breath condensate by flow injection analysis with fluorescence detection
A method for the determination of hydrogen peroxide in exhaled breath condensate (EBC) by automated flow injection analysis (FIA) with fluorescence detection was developed and validated. In the enzymatic assay a fluorescent dimer of para-hydroxyphenyl acetic acid (HPAA) was formed by the redox coupl...
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| Published in | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 809; no. 2; pp. 199 - 203 |
|---|---|
| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Elsevier B.V
05.10.2004
|
| Subjects | |
| Online Access | Get full text |
| ISSN | 1570-0232 1873-376X |
| DOI | 10.1016/j.jchromb.2004.06.027 |
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| Abstract | A method for the determination of hydrogen peroxide in exhaled breath condensate (EBC) by automated flow injection analysis (FIA) with fluorescence detection was developed and validated. In the enzymatic assay a fluorescent dimer of
para-hydroxyphenyl acetic acid (HPAA) was formed by the redox coupling reaction between hydrogen peroxide and horseradish peroxidase (HRP). The calibration curve of hydrogen peroxide was linear over a range of 40–5000
nM. The coefficient of variation (CV) for within-day precision was 1–3%; for between-day precision, it was 2–5% over the validated range. The assay requires a small sample aliquot (150
μl) and no incubation time, and has an analytical runtime of <2
min. It is therefore suitable for larger studies. The method was used to detect hydrogen peroxide in EBC of asthmatic patients and healthy volunteers. A statistically significant difference was found between patients with asthma (
n = 19) and control subjects without asthma (
n = 19), 780
nM versus 480
nM (
P = 0.03). |
|---|---|
| AbstractList | A method for the determination of hydrogen peroxide in exhaled breath condensate (EBC) by automated flow injection analysis (FIA) with fluorescence detection was developed and validated. In the enzymatic assay a fluorescent dimer of
para-hydroxyphenyl acetic acid (HPAA) was formed by the redox coupling reaction between hydrogen peroxide and horseradish peroxidase (HRP). The calibration curve of hydrogen peroxide was linear over a range of 40–5000
nM. The coefficient of variation (CV) for within-day precision was 1–3%; for between-day precision, it was 2–5% over the validated range. The assay requires a small sample aliquot (150
μl) and no incubation time, and has an analytical runtime of <2
min. It is therefore suitable for larger studies. The method was used to detect hydrogen peroxide in EBC of asthmatic patients and healthy volunteers. A statistically significant difference was found between patients with asthma (
n = 19) and control subjects without asthma (
n = 19), 780
nM versus 480
nM (
P = 0.03). A method for the determination of hydrogen peroxide in exhaled breath condensate (EBC) by automated flow injection analysis (FIA) with fluorescence detection was developed and validated. In the enzymatic assay a fluorescent dimer of para-hydroxyphenyl acetic acid (HPAA) was formed by the redox coupling reaction between hydrogen peroxide and horseradish peroxidase (HRP). The calibration curve of hydrogen peroxide was linear over a range of 40-5000 nM. The coefficient of variation (CV) for within-day precision was 1-3%; for between-day precision, it was 2-5% over the validated range. The assay requires a small sample aliquot (150 microl) and no incubation time, and has an analytical runtime of < 2 min. It is therefore suitable for larger studies. The method was used to detect hydrogen peroxide in EBC of asthmatic patients and healthy volunteers. A statistically significant difference was found between patients with asthma (n = 19) and control subjects without asthma (n = 19), 780 nM versus 480 nM (P = 0.03).A method for the determination of hydrogen peroxide in exhaled breath condensate (EBC) by automated flow injection analysis (FIA) with fluorescence detection was developed and validated. In the enzymatic assay a fluorescent dimer of para-hydroxyphenyl acetic acid (HPAA) was formed by the redox coupling reaction between hydrogen peroxide and horseradish peroxidase (HRP). The calibration curve of hydrogen peroxide was linear over a range of 40-5000 nM. The coefficient of variation (CV) for within-day precision was 1-3%; for between-day precision, it was 2-5% over the validated range. The assay requires a small sample aliquot (150 microl) and no incubation time, and has an analytical runtime of < 2 min. It is therefore suitable for larger studies. The method was used to detect hydrogen peroxide in EBC of asthmatic patients and healthy volunteers. A statistically significant difference was found between patients with asthma (n = 19) and control subjects without asthma (n = 19), 780 nM versus 480 nM (P = 0.03). |
| Author | Olin, Anna-Carin Ljungkvist, Göran Torén, Kjell Lärstad, Mona Svensson, Sophie |
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| Title | Determination of hydrogen peroxide in exhaled breath condensate by flow injection analysis with fluorescence detection |
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