Svensson Sophie, Olin Anna-Carin, Lärstad Mona, Ljungkvist Göran, Torén Kjell
Occupational and Environmental Medicine, Sahlgrenska University Hospital and Academy at Göteborg University, Sankt Sigfridsgatan 85, SE-41266 Göteborg, Sweden.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Oct 5;809(2):199-203. doi: 10.1016/j.jchromb.2004.06.027.
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).
开发并验证了一种通过自动流动注射分析（FIA）结合荧光检测来测定呼出气冷凝物（EBC）中过氧化氢的方法。在酶促测定中，过氧化氢与辣根过氧化物酶（HRP）之间的氧化还原偶联反应形成了对羟基苯乙酸（HPAA）的荧光二聚体。过氧化氢的校准曲线在40 - 5000 nM范围内呈线性。日内精密度的变异系数（CV）为1 - 3%；在验证范围内，日间精密度的变异系数为2 - 5%。该测定法所需样品量小（150微升）且无需孵育时间，分析运行时间< 2分钟。因此适用于更大规模的研究。该方法用于检测哮喘患者和健康志愿者的EBC中的过氧化氢。发现哮喘患者（n = 19）和无哮喘对照受试者（n = 19）之间存在统计学显著差异，分别为780 nM和480 nM（P = 0.03）。
