A mass spectrometric method to simultaneously measure a biomarker and dilution marker in exhaled breath condensate
A mass spectrometric method to simultaneously measure a biomarker and dilution marker in exhaled breath condensate
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DOI：
10.1002/rcm.3408
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发表时间：
2008-01-01
影响因子：
2
通讯作者：
Boysen, Gunnar
中科院分区：
文献类型：
--
作者：
Esther, Charles R., Jr.;Jasin, H. Matias;Boysen, Gunnar
呼出气冷凝液（EBC）采集是一种简单且无创的气道分泌物采样方法，但分析受限于冷凝液中气道分泌物的广泛和可变稀释。为了克服这一局限性，我们开发了一种灵敏和特异的液相色谱/串联质谱（LC/MS/MS）方法，同时检测腺苷嘌呤作为炎症的生物标志物和尿素作为EBC中的稀释标记物。使用C18色谱柱，以甲醇和甲酸作为移动的相，在质谱分析前实现分离，并监测特征前体至产物离子跃迁（m/z 268至136（对于腺苷）、m/z 348至136（对于AMP）和m/z 61至44（对于尿素））进行定量。为了校正基质效应，使用同位素标记的腺苷、AMP和尿素作为内标。使用这些方法，我们检测了尿素和腺苷嘌呤腺苷和AMP的EBC从7名受试者囊性纤维化（CF）和7名健康对照，发现AMP/尿素的比例升高CF样品。这些结果表明，质谱法可以成功地用于EBC分析，以同时检测气道炎症的生物标志物和可变稀释的对照。版权所有（C）2008约翰威利父子有限公司
Exhaled breath condensate (EBC) collection is a simple and non-invasive method to sample airway secretions, but analysis is limited by extensive and variable dilution of airway secretions within the condensate. To overcome this limitation, we developed a sensitive and specific liquid chromatography/tandem mass spectrometry (LC/MS/MS) method to simultaneously detect adenyl purines as biomarkers of inflammation and urea as a dilution marker in EBC. Separation prior to mass spectrometry was achieved using a C18 column with methanol and formic acid as the mobile phase, and characteristic precursor to product ion transitions of m/z 268 to 136 (for adenosine), m/z 348 to 136 (for AMP), and m/z 61 to 44 (for urea) were monitored for quantification. To correct for matrix effects, isotopically labeled adenosine, AMP, and urea were used as internal standards. Using these methods, we detected urea and the adenyl purines adenosine and AMP in EBC from seven subjects with cystic fibrosis (CF) and seven healthy controls and found that the AMP/urea ratio was elevated in the CF samples. These results demonstrate that mass spectrometry can be used successfully in EBC analysis to simultaneously detect a biomarker for airway inflammation and control for variable dilution. Copyright (C) 2008 John Wiley & Sons, Ltd.