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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >Quantification of free and total desmosine and isodesmosine in human urine by liquid chromatography tandem mass spectrometry: A comparison of the surrogate-analyte and the surrogate-matrix approach for quantitation
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Quantification of free and total desmosine and isodesmosine in human urine by liquid chromatography tandem mass spectrometry: A comparison of the surrogate-analyte and the surrogate-matrix approach for quantitation

机译：液相色谱-串联质谱法定量测定人尿中游离和总地高硫代和异高铁素的含量：替代分析物和替代基质法的定量比较

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摘要

In spite of the data suggesting the potential of urinary desmosine (DES) and isodesmosine (IDS) as biomarkers for elevated lung elastic fiber turnover, further validation in large-scale studies of COPD populations, as well as the analysis of longitudinal samples is required. Validated analytical methods that allow the accurate and precise quantification of DES and IDS in human urine are mandatory in order to properly evaluate the outcome of such clinical studies. In this work, we present the development and full validation of two methods that allow DES and IDS measurement in human urine, one for the free and one for the total (free + peptide-bound) forms. To this end we compared the two principle approaches that are used for the absolute quantification of endogenous compounds in biological samples, analysis against calibrators containing authentic analyte in surrogate matrix or containing surrogate analyte in authentic matrix. The validated methods were employed for the analysis of a small set of samples including healthy never-smokers, healthy current-smokers and COPD patients. This is the first time that the analysis of urinary free DES, free IDS, total DES, and total IDS has been fully validated and that the surrogate analyte approach has been evaluated for their quantification in biological samples. Results indicate that the presented methods have the necessary quality and level of validation to assess the potential of urinary DES and IDS levels as biomarkers for the progression of COPD and the effect of therapeutic interventions.
机译：尽管有数据表明尿液中的地塞米辛（DES）和异地西斯莫辛（IDS）有可能成为提高肺弹性纤维更新率的生物标志物，但仍需要在大规模COPD人群研究以及纵向样品分析中进行进一步验证。为了正确评估此类临床研究的结果，必须使用经过验证的分析方法，才能对人尿中的DES和IDS进行准确和精确的定量。在这项工作中，我们介绍了两种允许在人尿中进行DES和IDS测量的方法的开发和全面验证，一种用于游离形式，一种用于总形式（游离+肽结合）。为此，我们比较了两种绝对方法，用于对生物样品中的内源性化合物进行绝对定量，针对在替代基质中包含真实分析物或在真实基质中包含替代分析物的校准物进行分析。经过验证的方法用于分析一小部分样本，包括健康从未吸烟者，健康目前吸烟者和COPD患者。这是首次完全验证了尿中游离DES，游离IDS，总DES和总IDS的分析，并且已经评估了替代分析物方法在生物样品中的定量分析。结果表明，提出的方法具有必要的质量和验证水平，以评估尿DES和IDS水平作为COPD进展和治疗干预效果的生物标志物的潜力。

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