尿液代谢组学非靶向质谱分析法两种采集模式的比较

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (5): 729-734.

尿液代谢组学非靶向质谱分析法两种采集模式的比较

肖晓莲¹, 刘晓燕¹, 朱文凤², 杨晔宏², 杨俊涛², 孙伟^1*

中国医学科学院基础医学研究所北京协和医学院基础学院 1.药理系; 2.医学分子生物学国家重点实验室,北京 100005

收稿日期:2021-01-08 修回日期:2021-03-20 出版日期:2021-05-05 发布日期:2021-05-06
通讯作者: ^*sunwei@ibms.pumc.edu.cn
基金资助:
中国医学科学院医学科学创新基金(2017-I2M-1-009)

Comparison of two acquisition modes in urine metabolomics by untargeted mass spectrometry

XIAO Xiao-lian¹, LIU Xiao-yan¹, ZHU Wen-feng², YANG Ye-hong², YANG Jun-tao², SUN Wei^1*

1. Department of Pharmacology; 2. State Key Labolatory of Medical Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC,Beijing 100005,China

Received:2021-01-08 Revised:2021-03-20 Online:2021-05-05 Published:2021-05-06
Contact: ^*sunwei@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的比较全扫描采集(FSA)模式和数据依赖采集(DDA)模式用于非靶向代谢组分析的优缺点。方法用代谢物标准品混合物和健康人尿液样品分别进行两种模式的质谱采集,比较两种模式的定性和定量效果。结果在标准品代谢物分析时,FSA模式下的标准品代谢物丰度比DDA模式平均高2.36倍(P＜0.05),FSA、DDA组的丰度变异系数(CV)范围分别为0.81%～6.35％和2.14%～11.20％。在进行尿液样品分析时,FSA模式的谱图数和代谢特征数量与DDA模式差异无统计学意义。不同进样量(1、2和4 μL)时,FSA模式鉴定的代谢物数量比DDA模式多(分别高于9.5％、1.9％和10％),FSA模式代谢物定量CV中位数(分别为7.1％、3.0％和2.6％)明显低于DDA模式者(分别为10.3％、7.0％和6.5％)。进样量1、2和4 μL时,随着代谢物丰度的升高,DDA的CV均数较FSA组分别高1.3～1.7、1.7～1.8和1.4～1.9倍(P＜0.05);而且,随着峰宽增加,DDA组的CV均数较FSA组分别高1.5～1.8、1.7～2.1和2.0～2.2倍(P＜0.05)。结论 FSA模式对系统稳定性要求高,更适合短时间小规模样品量分析;DDA模式对系统稳定性要求相对较低,操作简便省时,可用于长时间大规模样品量的分析。

关键词: 非靶向代谢组学, 全扫描采集(FSA), 数据依赖采集(DDA)

Abstract: Objective To compare the advantages and disadvantages of full scan acquisition (FSA) mode and data dependent acquisition (DDA) mode for untargeted metabolomic analysis. Methods Qualitative and quantitative results of two acquisition methods were compared by using a standard mixture of 9 metabolites and human urine sample. Results In the analysis of standard sample, the metabolite abundance in FSA mode was about 2.36 times higher than that in DDA mode (P＜0.05), the CV of metabolites abundance ranged from 2.14% to 11.20% in DDA mode and from 0.81% to 6.35% in FSA mode. In the analysis of urine sample, there was no statistical difference in the number of spectra and metabolite feature between FSA mode and DDA mode. When injecting 1, 2 and 4 μL, the number of metabolites identified by FSA mode was 9.5％, 1.9％ and 10％ higher than those identified by DDA mode, respectively. And the median CV of metabolite abundance analyzed by FSA(7.1％,3.0％ and 2.6％)was significantly lower than that by DDA(10.3％, 7.0% and 6.5%). When injecting 1, 2 and 4 μL,with the increase of abundance, the median CV of metabolite abundance in DDA mode was 1.3-1.7,1.7-1.8 and 1.4-1.9 times higher than that in FSA mode (P＜0.05), and with the increase of peak width, the median CV of DDA mode was 1.5-1.8, 1.7-2.1 and 2.0-2.2 times higher than that of FSA mode (P＜0.05). Conclusions FSA mode requires high system stability, it is suitable for fast and small-scale sample analysis; DDA mode has lower requirement for system stability, and is convenient and time-saving, which can be used for long-time and large-scale sample analysis.

Key words: untargeted metabolomics, full scan acquisition(FSA), data dependent acquisition(DDA)

中图分类号:

R331

肖晓莲, 刘晓燕, 朱文凤, 杨晔宏, 杨俊涛, 孙伟. 尿液代谢组学非靶向质谱分析法两种采集模式的比较[J]. 基础医学与临床, 2021, 41(5): 729-734.

XIAO Xiao-lian, LIU Xiao-yan, ZHU Wen-feng, YANG Ye-hong, YANG Jun-tao, SUN Wei. Comparison of two acquisition modes in urine metabolomics by untargeted mass spectrometry[J]. Basic & Clinical Medicine, 2021, 41(5): 729-734.

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