A high throughput lipidomics method and its application in atrial fibrillation based on 96-well plate pretreatment and liquid chromatography-mass spectrometry.
A high throughput lipidomics method and its application in atrial fibrillation based on 96-well plate pretreatment and liquid chromatography-mass spectrometry.
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DOI：
10.1016/j.chroma.2021.462271
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发表时间：
2021-05
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通讯作者：
Yuqing Zhang;Yun-jie Xie;Wangjie Lv;Chunxiu Hu;Tianrun Xu;Xinyu Liu;Rongfeng Zhang;Guowang Xu;Yunlong Xia;Xinjie Zhao
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作者：
Yuqing Zhang;Yun-jie Xie;Wangjie Lv;Chunxiu Hu;Tianrun Xu;Xinyu Liu;Rongfeng Zhang;Guowang Xu;Yunlong Xia;Xinjie Zhao
脂质组学在临床上的成功应用需要对大规模样品进行研究，而脂质分析方法的通量和稳健性是其瓶颈。在这里，我们报告了一种非靶向脂质组学方法，通过将96孔板中的高通量预处理与四极杆飞行时间串联质谱联用的超高效液相色谱相结合。经验证，所开发的方法在线性、重复性和提取回收率方面具有令人满意的分析特性。该仪器可在25 min内同时处理96份样品，检测血浆样品中441种脂类。脂质提取物的储存稳定性研究为大规模样品分析提供了可操作的方法，并证明大多数脂质在预处理后10 °C下36 h内和-80 °C下72 h内在自动进样器中稳定。为了证明该方法的有效性，采用该方法来研究与心房颤动相关的血浆脂质体异常。在发现集和外部验证集中分别获得了曲线下面积（AUC）值为0.831和0.745的生物标志物组。这些结果表明，所开发的方法适用于大规模生物样品处理和血浆脂质分析。
Successful applications of lipidomics in clinic need study large-scale samples, and the bottlenecks are in throughput and robustness of the lipid analytical method. Here, we report an untargeted lipidomics method by combining high throughput pretreatment in the 96-well plate with ultra-high performance liquid chromatography coupled to quadrupole time-of-flight tandem mass spectrometry. The developed method was validated to have satisfactory analytical characteristics in terms of linearity, repeatability and extraction recovery. It can be used to handle 96 samples simultaneously in 25 min and detect 441 lipids in plasma sample. Storage stability investigation on lipid extracts provided an operable procedure for large-scale sample analysis and demonstrated most lipids were stable in autosampler at 10 °C within 36 h and at -80 °C within 72 h after the pretreatment. To prove the usefulness, the method was employed to investigate abnormal plasma lipidome related to atrial fibrillation. A biomarker panel with the area under the curve (AUC) values of 0.831 and 0.745 was achieved in the discovery and external validation sets, respectively. These results showed that the developed method is applicable for large-scale biological sample handling and lipid analysis of plasma.