尿液代谢组学研究中质控样本的制备方法研究

目的初步探索代谢组学研究中尿液质控样本的制备方法。方法运用液质联用技术对健康志愿者、镉暴露者尿样和质控样本检测分析,并对获得的总离子流图、内源性代谢物含量及主成分进行比较。结果该研究样本为40名女性健康志愿者晨尿、40名女性镉暴露者晨尿及来自于80份样本等体积混匀配置成的18份质控样本。随机选取的6份健康志愿者和6份镉暴露者尿样在正、负离子检测模式条件下总离子流图不能完全重叠,6份质控样本总离子流图几乎完全重合。内源性代谢物log2转化的归一化丰度值的四分位数间距比较结果显示,质控样本与健康志愿者和镉暴露者样本代谢物log2转化的归一化丰度值分布之间差异均有统计学意义（均P〈0.05）;主成分分析（PCA）得分图显示健康志愿者及镉暴露者样本散在分布,而质控样相对集中;进一步对质控样本分析发现,除平衡系统时使用的5个质控样本之外,其他13个质控样本变化均保持在2SD之间。结论该研究配置的质控样本能够反映研究样本基本信息及系统的受控情况,建立了适用于尿液代谢组学研究的质控样本制备方法。

Study on preparing quality control samples for urine metabonomics research

Objective To explore the method of preparing urine quality control samples in metabonomics research. Methods Urine samples collected from healthy volunteers and cadmium exposure in- dividuals, and quality control samples were analyzed by LC/MS techniques. Then, the total ion currents, the content of endogenous metabolites and principal components were compared. Results Study samples consisted of 40 morning urine samples from female healthy volunteers, 40 morning urine samples from cad- mium exposure females, and 18 quality control samples prepared by mixing 80 urine research samples at e- qual volume from 40 morning urine samples of female healthy volunteers and 40 morning urine samples of cadmium exposure females. The total ion currents of 6 samples randomly selected from healthy volunteers and 6 morning urine samples randomly from cadmium exposure females were disordered, but 6 quality con- trol samples nearly overlapped in EST ＋/- conditions. The inter-quartile range of endogenous metabolites indicated that contents of quality control samples were significantly different from those of healthy volunteers and cadmium exposure females （ both P 〈 0. 05 ）. The score plots of principal component analysis （PCA） showed that urine samples of healthy volunteers and cadmium exposure females distributed dispersedly, but quality control samples concentrated. Except five quality control samples used in balance system, the deviation of the other thirteen quality control samples was kept in 2SD. Conclusion Quality control samples prepared in this study could reflect the basic information of samples and the status of system control, and the method of preparing quality control samples in urine metabonomics research was established.