Effects of sample processing, time and storage condition on cysteinyl leukotrienes in exhaled breath condensate.
Effects of sample processing, time and storage condition on cysteinyl leukotrienes in exhaled breath condensate.
复制标题
样品处理、时间和储存条件对呼出气冷凝液中半胱氨酰白三烯的影响。
DOI：
10.1088/1752-7155/4/4/046002
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发表时间：
2010
影响因子：
3.8
通讯作者：
Debley,JasonS
中科院分区：
文献类型：
--
作者：
Ohanian,ArpyS;Zimmerman,Jerry;Debley,JasonS
可以在呼出气冷凝液 (EBC) 中测量半胱氨酰白三烯 (CysLT)；然而，已发表的研究中，哮喘受试者和健康受试者报告的 EBC CysLT 浓度存在相当大的差异，这可能部分是由于加工和储存过程中 CysLT 降解所致。我们评估了健康受试者在 EBC 中 6 个月内的 CysLT 稳定性，这些受试者在−80°C 下保存，用氩气分层，然后在−80°C 下保存，或在固相萃取 (SPE) 后在−80°C 下保存在 0.2% 甲酸的甲醇溶液中。我们发现，在储存于− 80°C 或用氩气分层的情况下，加标和未加标的 EBC 样品中的 CysLT 随时间显着降解。 SPE 后，在 0.2% 甲酸的甲醇溶液中储存 6 个月后，CysLT 回收率显着提高；然而，随着时间的推移，内源性 CysLT 回收率存在显着差异，这可能归因于 CysLT 测定范围低端的测定间和测定内差异。尽管在 SPE 后储存在甲醇中的 EBC 中 CysLT 的回收率更高，但这种方法引入的变异程度似乎高得令人无法接受。我们认为，在 CysLT 能够可靠地用作呼出气中的生物标志物之前，需要开发更灵敏、变量更少的方法来量化 EBC 中的 CysLT。在设计临床研究（包括评估作为生物标志物的 EBC 成分）时，应仔细考虑样品处理和储存以及测定间和测定内的变异性。
Cysteinyl leukotrienes (CysLTs) can be measured in exhaled breath condensate (EBC); however, there is considerable variation in reported EBC CysLT concentrations from asthmatic and healthy subjects between published studies, which may be partially explained by CysLT degradation during processing and storage. We assessed CysLT stability over 6 months in EBC from healthy subjects stored at− 80 C, layered with argon and then stored at− 80 C or stored in 0.2% formic acid in methanol at− 80 C following solid-phase extraction (SPE). We found significant CysLT degradation over time in both spiked and unspiked EBC samples stored at− 80 C or layered with argon. CysLT recovery was significantly greater after storage for 6 months in 0.2% formic acid in methanol following SPE; however, there was substantial variability in endogenous CysLT recovery over time, possibly attributable to inter-and intra-assay variability at the low end of the CysLT assay range. Despite the greater recovery of CysLTs in EBC stored in methanol following SPE, the degree of variability introduced by this method appears unacceptably high. We believe that the development of more sensitive and less variable methods for quantifying CysLTs in EBC are required before CysLTs can reliably be utilized as biomarkers in exhaled breath. Sample processing and storage, as well as inter-and intra-assay variability, should be carefully considered in the design of clinical studies that include assessments of EBC constituents as biomarkers.