用于定量血浆和血清中总去甲二甲基赖氨酰氧化酶和异去甲二甲基赖氨酰氧化酶的同位素稀释液相色谱-串联质谱法的表征与验证
Characterization and validation of an isotope-dilution LC-MS/MS method for quantification of total desmosine and isodesmosine in plasma and serum.
作者信息
Albarbarawi Osama, Barton Alun, Miller Douglas, McSharry Charles, Chaudhuri Rekha, Thomson Neil C, Palmer Colin Na, Devereux Graham, Huang Jeffrey T-J
机构信息
Translational Medicine Research Collaboration, TMRC Laboratory, James Arrott Drive, Dundee, DD1 9SY, UK.
出版信息
Bioanalysis. 2013 Aug;5(16):1991-2001. doi: 10.4155/bio.13.164.
BACKGROUND
Desmosine/isodesmosine (DES/IDS) is a promising biomarker for estimating activity of elastin degradation. RESULTS/METHODOLOGY: A stable isotope dilution LC-MS/MS method for measuring serum/plasma DES/IDS was developed and validated. The reportable range of this assay was 0.1-160 ng/ml. Serum/plasma DES/IDS level was stable at room temperature or 4°C for 20 h, and for three freeze-thaw cycles. Interferences from endogenous compounds and ion suppression/enhancing effect were also evaluated. Our results suggest the absolute necessity of using an IS in the measurement. We found that serum/plasma DES/IDS levels from patients with chronic obstructive pulmonary disease and cystic fibrosis were significantly higher compared with healthy smokers.
CONCLUSION
These results demonstrate that the LC-MS/MS method provides sensitive, reproducible and accurate quantification of total serum/plasma DES/IDS.
背景
锁链素/异锁链素（DES/IDS）是一种很有前景的用于评估弹性蛋白降解活性的生物标志物。
结果/方法：开发并验证了一种用于测量血清/血浆中DES/IDS的稳定同位素稀释液相色谱-串联质谱法。该检测方法的报告范围为0.1-160 ng/ml。血清/血浆DES/IDS水平在室温或4°C下20小时内以及经过三个冻融循环后保持稳定。还评估了内源性化合物的干扰以及离子抑制/增强效应。我们的结果表明在测量中使用内标物是绝对必要的。我们发现慢性阻塞性肺疾病和囊性纤维化患者的血清/血浆DES/IDS水平明显高于健康吸烟者。
结论
这些结果表明液相色谱-串联质谱法能对血清/血浆中的总DES/IDS进行灵敏、可重复且准确的定量分析。
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