van der Laan Tom, Boom Isabelle, Maliepaard Joshua, Dubbelman Anne-Charlotte, Harms Amy C, Hankemeier Thomas
Analytical Biosciences and Metabolomics, Division of Systems Biomedicine and Pharmacology, Leiden Academic Center for Drug Research, Leiden University, 2333 CC Leiden, The Netherlands.
Metabolites. 2020 Dec 18;10(12):514. doi: 10.3390/metabo10120514.
A popular fragmentation technique for non-targeted analysis is called data-independent acquisition (DIA), because it provides fragmentation data for all analytes in a specific mass range. In this work, we demonstrated the strengths and weaknesses of DIA. Two types of chromatography (fractionation/3 min and hydrophilic interaction liquid chromatography (HILIC)/18 min) and three DIA protocols (variable sequential window acquisition of all theoretical mass spectra (SWATH), fixed SWATH and MS) were used to evaluate the performance of DIA. Our results show that fast chromatography and MS often results in product ion overlap and complex MS/MS spectra, which reduces the quantitative and qualitative power of these DIA protocols. The combination of SWATH and HILIC allowed for the correct identification of 20 metabolites using the NIST library. After SWATH window customization (i.e., variable SWATH), we were able to quantify ten structural isomers with a mean accuracy of 103% (91-113%). The robustness of the variable SWATH and HILIC method was demonstrated by the accurate quantification of these structural isomers in 10 highly diverse blood samples. Since the combination of variable SWATH and HILIC results in good quantitative and qualitative fragmentation data, it is promising for both targeted and untargeted platforms. This should decrease the number of platforms needed in metabolomics and increase the value of a single analysis.
一种用于非靶向分析的常用碎片化技术称为数据非依赖采集（DIA），因为它能为特定质量范围内的所有分析物提供碎片化数据。在这项工作中，我们展示了DIA的优缺点。使用了两种色谱法（分馏/3分钟和亲水作用液相色谱法（HILIC）/18分钟）以及三种DIA方案（所有理论质谱的可变顺序窗口采集（SWATH）、固定SWATH和MS）来评估DIA的性能。我们的结果表明，快速色谱法和MS常常导致产物离子重叠以及复杂的二级质谱图，这降低了这些DIA方案的定量和定性能力。SWATH与HILIC的结合使得使用NIST库能够正确鉴定20种代谢物。在对SWATH窗口进行定制（即可变SWATH）后，我们能够对十种结构异构体进行定量，平均准确度为103%（91 - 113%）。通过在10种高度多样化的血液样本中对这些结构异构体进行准确量化，证明了可变SWATH和HILIC方法的稳健性。由于可变SWATH和HILIC的结合能产生良好的定量和定性碎片化数据，它在靶向和非靶向平台上都很有前景。这应该会减少代谢组学所需的平台数量，并提高单次分析的价值。
