Pre-analytical sample quality: metabolite ratios as an intrinsic marker for prolonged room temperature exposure of serum samples.
Pre-analytical sample quality: metabolite ratios as an intrinsic marker for prolonged room temperature exposure of serum samples.
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DOI：
10.1371/journal.pone.0121495
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Waldenberger M
中科院分区：
文献类型：
--
作者：
Anton G;Wilson R;Yu ZH;Prehn C;Zukunft S;Adamski J;Heier M;Meisinger C;Römisch-Margl W;Wang-Sattler R;Hveem K;Wolfenbuttel B;Peters A;Kastenmüller G;Waldenberger M
“组学”领域的进步带来了对大量高质量样品的需求。许多组学研究利用生物库样本来满足这一需求。大多数实验室错误发生在分析前阶段。因此，迫切需要用于预分析阶段的基于证据的标准操作程序以及区分“好”和“坏”质量样品的标记，同时考虑到所需的下游分析。我们研究了由于储存前处理条件以及由于重复冻融循环引起的血清样品中代谢物的浓度变化。我们采集了空腹血清样本，并在室温（RT）和湿冰上和干冰上对等分试样进行了最多4次冻融循环和12、24和36小时的储存前处理延迟。对于每个处理的等分试样，我们通过靶向代谢组学方法定量了127种代谢物。我们发现在室温下保存的样品中有明显的降解特征。在湿冰上储存导致浓度变化不太明显。24种代谢产物在RT下显示出显著的浓度变化。其中22种代谢产物在储存延迟仅12小时后就已出现变化。尤其明显的是溶血磷脂酰胆碱增加和磷脂酰胆碱减少。我们表明，这些分子类别的浓度之间的比率可以作为一个衡量标准，以区分“好”和“坏”的质量样品在我们的研究。相比之下，我们发现在多达四个冻融循环期间代谢物浓度相当稳定。我们得出结论，应严格避免血清样品的分析前RT处理，血清样品应始终在湿冰上或离心后在冷却装置中处理。此外，血清样本应在离心后尽快在-80 ° C或以下冷冻。
Advances in the “omics” field bring about the need for a high number of good quality samples. Many omics studies take advantage of biobanked samples to meet this need. Most of the laboratory errors occur in the pre-analytical phase. Therefore evidence-based standard operating procedures for the pre-analytical phase as well as markers to distinguish between ‘good’ and ‘bad’ quality samples taking into account the desired downstream analysis are urgently needed. We studied concentration changes of metabolites in serum samples due to pre-storage handling conditions as well as due to repeated freeze-thaw cycles. We collected fasting serum samples and subjected aliquots to up to four freeze-thaw cycles and to pre-storage handling delays of 12, 24 and 36 hours at room temperature (RT) and on wet and dry ice. For each treated aliquot, we quantified 127 metabolites through a targeted metabolomics approach. We found a clear signature of degradation in samples kept at RT. Storage on wet ice led to less pronounced concentration changes. 24 metabolites showed significant concentration changes at RT. In 22 of these, changes were already visible after only 12 hours of storage delay. Especially pronounced were increases in lysophosphatidylcholines and decreases in phosphatidylcholines. We showed that the ratio between the concentrations of these molecule classes could serve as a measure to distinguish between ‘good’ and ‘bad’ quality samples in our study. In contrast, we found quite stable metabolite concentrations during up to four freeze-thaw cycles. We concluded that pre-analytical RT handling of serum samples should be strictly avoided and serum samples should always be handled on wet ice or in cooling devices after centrifugation. Moreover, serum samples should be frozen at or below -80°C as soon as possible after centrifugation.
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影响因子：
3
作者：
Petersen AK;Krumsiek J;Wägele B;Theis FJ;Wichmann HE;Gieger C;Suhre K
通讯作者：
Suhre K
存储时间和冻融周期对血清样品稳定性的影响。
影响因子：
3.3
作者：
Cuhadar S;Koseoglu M;Atay A;Dirican A
通讯作者：
Dirican A
影响因子：
3.7
作者：
Breier M;Wahl S;Prehn C;Fugmann M;Ferrari U;Weise M;Banning F;Seissler J;Grallert H;Adamski J;Lechner A
通讯作者：
Lechner A
影响因子：
30.8
作者：
通讯作者：
--
影响因子：
7.7
作者：
Clark, S;Youngman, LD;Collins, R
通讯作者：
Collins, R