色谱保留时间校正在非靶向筛查中的应用

Application of chromatographic retention time correction in untargeted screening

目的色谱保留时间校正是有效提高鉴定准确性的重要步骤之一。本文提出了一种基于保留时间校正的生物样本非靶向筛查的策略。方法建立生物样本前处理方法,优化液相色谱条件、质谱检测条件,筛选14个化合物作为校正剂,考察不同样品、不同进样时间、不同品牌仪器、更换色谱柱、更换流动相后保留时间校正情况。结果校正剂覆盖范围广、稳定性好且不干扰样本测定,在正、负离子模式下能够均匀分布在色谱图中,将色谱图分割为若干时间段,每个时间段内使用校正剂进行保留时间线性校正,校正效果良好。结论引入保留时间校正方法后可消除实验条件造成的保留时间漂移,提高定性的准确度,有助于解决仅依据质谱信息定性的结果假阳性高的问题。

OBJECTIVE Chromatographic retention time correction is one of the important steps to effectively improve the accuracy of identification.This article proposed a strategy for untargeted screening of biological samples based on retention time correction.METHODS A pre-treatment method for biological samples was established.The conditions of liquid chromatography and mass spectrometry were optimized.Fourteen compounds were selected as calibration agents.The retention time correction of different samples,different injection time,different brands of instruments,changing chromatographic column and changing mobile phase were investigated.RESULTS Calibration agents had a wide coverage,good stability and no interference with sample determination.They could be uniformly distributed in the chromatogram in both positive and negative ion modes.The chromatogram was divided into several time intervals.Calibration agents in each time period were used for retention time linear correction,and the correction effects were good.CONCLUSION The retention time correction method could eliminate the retention time drift caused by experimental conditions,improve the accuracy of qualitative analysis,and help to solve the problem of high false positive result based on mass spectrum information.