Evaluation of nanospray capillary LC-MS performance for metabolomic analysis in complex biological matrices
Evaluation of nanospray capillary LC-MS performance for metabolomic analysis in complex biological matrices
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DOI：
10.1016/j.chroma.2022.462952
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发表时间：
2022-03-23
影响因子：
4.1
通讯作者：
Mostafa, Mahmoud Elhusseiny
中科院分区：
文献类型：
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作者：
Grinias, James P.;Edwards, James L.;Mostafa, Mahmoud Elhusseiny
使用大孔径色谱柱时，复杂生物基质中的 LC-MS 代谢组分析可能会因简并性而变得复杂。简并性是指由于盐与代谢物的加合、二聚化或中性物的丢失而从同一分析物中检测到多个质谱峰。这会对 MS 谱图产生干扰，减少定量，并增加错误识别率。使用 2.1 mm 内径 (i.d.) 色谱柱进行分析通常会导致简并峰，而使用毛细管柱（25、50 和 75 μm i.d.）的纳喷雾可减少简并峰。针对氨基酸标准品的毛细管 LC 色谱参数的优化显示，所有毛细管内径的 HETP 最低为 1.25 毫米/秒。列。结果表明质量敏感检测低于最佳速度。以更快的速度，所有毛细血管中都会发生浓度依赖性检测。内径 2.1 毫米分析级色谱柱显示出最大程度的简并性，特别是在低信号强度范围内。内径 25 微米对于相同的信号强度范围，柱显示出更高水平的代谢物注释。它还提供了最低水平的简并性，使其最适合非目标分析。内径 25 微米在 30 分钟梯度法中，色谱柱的峰值容量 (n(c)) 为 144，其中 nc 随着色谱柱内径的增加而减小。增加。内径 75 微米毛细管柱显示出最高的信号强度，这有利于靶向分析。比较了毛细管和分析规模色谱柱的色谱性能、分辨率和简并性特征的这些影响，以用于复杂血清和细胞裂解物基质的代谢组学分析。 (C) 2022 Elsevier B.V. 保留所有权利。
LC-MS metabolomic analysis in complex biological matrices may be complicated by degeneracy when using large-bore columns. Degeneracy is the detection of multiple mass spectral peaks from the same analyte due to adduction of salts to the metabolite, dimerization, or loss of neutrals. This introduces interferences to the MS spectra, diminishes quantification, and increases the rate of false identifications. Analysis using 2.1 mm inner diameter (i.d.) columns typically leads to degenerate peaks whereas nanospray using capillary columns (25, 50, and 75 mu m i.d.) reduces degeneracy. Optimization of chromatographic parameters of capillary LC for amino acid standards showed the lowest HETP at 1.25 mm/sec across all capillary i.d. columns. Results suggest mass-sensitive detection below the optimum velocity. At faster velocities, concentration-dependent detection occurred across all capillaries. The 2.1 mm i.d. analytical scale column showed the greatest level of degeneracy, particularly in the low signal intensity range. 25 mu m i.d. columns showed higher levels of metabolite annotation for the same signal intensity range. It also provided the lowest level of degeneracy, making it best suited for untargeted analysis. The 25 mu m i.d. column achieved a peak capacity (n(c)) of 144 in a 30-minute gradient method with nc decreasing as the column i.d. increased. 75 mu m i.d. capillary columns showed the highest signal intensity, which is beneficial for targeted analysis. These effects of chromatographic performance, resolution, and degeneracy profile of capillary and analytical scale columns were compared for metabolomic analyses in complex serum and cell lysate matrices. (C) 2022 Elsevier B.V. All rights reserved.