不同储存条件下人尿中小分子代谢物表征差异的初步研究

Primary exploration of the difference of micromolecular metabolites in urine under different storage conditions

目的了解代谢组学研究中常用的4种尿液储存条件下人尿中小分子代谢物表征的差异。方法采用超高效液相色谱串联飞行时间质谱(UPLC-Q/TOF),分别在正离子和负离子模式下检测5名志愿者尿液中小分子代谢物,运用多元统计法分析4种储存条件(-80℃、-20℃、1%HNO3置-80℃和2%抗坏血酸置-80℃)影响代谢物表征的差异性。结果方差分析显示4种常见方法保存后的尿液检测到1375个具有显著性差异的小分子代谢物(ESI+/-为613/762个,P<0.001),修正后,-80℃组尿液中存在小分子代谢物3700多个,与-80℃组相比,-20℃组存在4个具有显著性差异的小分子代谢物(ESI+/-为2/2个),1%HNO3置-80℃保存后尿液中有120个(ESI+/-为76/44个),2%抗坏血酸置-80℃保存尿液中有18个(ESI+/-为6/12个)。结论本研究通过对比尿液的4种保存条件后尿液中小分子代谢物表征差异,初步认定-80℃为首选保存条件。

Objective To know the difference of small molecular metabolites characteristics in urine under 4 common storage conditions. Methods Using UPLC - Q/TOF, urine micromolecular metabolites were detected in 5 volunteers in the positive and negative ion mode, and multivariate statistic analysis was employed to explore the different characteristics of urine micromolecular metabolites under 4 storage conditions, - 80 ℃ , - 20 ℃ , 1% HNO3 - 80 ℃ and 2% ascorbie acid - 80 ℃. Results One - way ANOVA showed that there were 1375 kinds of micromoleeules metabolites with significant difference among 4 storage conditions （ ESI ＋ / - were 613/762, P 〈 0.001 ）. After correction, there were more than 3700 kinds of micromolecule metabolites in urine stored at - 80 ℃. Compared with the storage condition of - 80 ℃, there were 4 micromolecular metabolites with significant difference at -20 ℃ （ESI ＋/- were 2/2） , 120 micromolecular metabolites with significant difference （ESI ＋/- were 76/44） in 1% HNO3 -g0 ℃ and lg micromolecular metabolites with significant difference （ESI ＋/- were 6/12） in 2% ascorbic acid -80 ℃. Conclusion By comparing the urine micromolecule metabolites characteristics under 4 storage conditions, -80 ℃ was regarded as the preferred storage condition.