Implementing 1.5 mm internal diameter columns into analytical workflows
Implementing 1.5 mm internal diameter columns into analytical workflows
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在分析工作流程中实施 1.5 mm 内径色谱柱
DOI：
10.1016/j.chroma.2022.463207
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发表时间：
2022
影响因子：
4.1
通讯作者：
Boyes, Barry E.
中科院分区：
文献类型：
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作者：
Libert, Benjamin P.;Godinho, Justin M.;Foster, Samuel W.;Grinias, James P.;Boyes, Barry E.
在液相色谱（LC）中使用较小的柱直径通常与毛细管LC相关联。虽然通过调整这种格式获得了许多分析益处，但由于柱易碎和额外的柱分散，常规使用仍然具有挑战性。桥接常规使用的2.1 mm柱和毛细管孔柱之间的差距允许连续的但远非微不足道的性能增加，而不需要与非常低分散LC系统相关的专用设备。此外，柱内径（i.d.）允许类似的质量负载（避免柱过载，柱过载可在内径的更大减小中观察到）。而没有陷获），从而增加了测量信号。因此，1.5mm i.d.柱提供了在更常用的2.1mm内径之间的可选择的中间尺寸。柱和Imm i.d.列.与2.1 mm i.d.相比，这些色谱柱平衡了灵敏度的增加。柱（对于相同的质量负载，理论上使质量敏感检测器中的时域峰面积加倍），同时减轻由于柱外分散效应而导致的效率损失，所述柱外分散效应通常在1.0 mm i.d.列.在此，使用1.5mm i.色谱柱应用于小分子的LC/UV分析和单克隆抗体的LC/MS分析。在柱上具有等效质量负载的情况下，1.5 mm i.d.对于小分子以及完整、亚基和肽水平的抗体分析，柱提供分析物峰面积信号的2 - 3倍改进。使用1.5 mm i.d.柱，尽管增加的规模在等度和梯度模式之间变化，可能是由于系统分散效应的差异和不同流速下电喷雾电离效率的变化。
The use of smaller column diameters in liquid chromatography (LC) is often associated with capillary LC. Although there are many analytical benefits gained by adapting this format, routine use continues to be challenging due to column fragility and extra column dispersion. Bridging the gap between routinely used 2.1 mm columns and capillary bore columns allows for a sequential but far from insignificant increase in performance without the need for specialized equipment associated with very low dispersion LC systems. Moreover, an incremental decrease in column internal diameter (i.d.) allows for similar mass load (avoiding column overload that may be observed in much larger decreases in i.d. without trapping) and thus an increase in measured signal. As such, 1.5 mm i.d. columns provide an alternative intermediate dimension between the more regularly used 2.1 mm i.d. columns and 1 mm i.d. columns. These columns balance an increase in sensitivity compared to 2.1 mm i.d. columns (theoretically doubling the time-domain peak area in mass sensitive detectors for the same mass load), while mitigating the efficiency losses due to extra-column dispersion effects that are commonly observed with 1.0 mm i.d. columns. Here, the use of 1.5 mm i.d. columns was applied to LC/UV analysis of small molecules and LC/MS methods for the analysis of monoclonal antibodies. With equivalent mass load on column, the 1.5 mm i.d. columns provide two-to-threefold improvement in analyte peak area signal for small molecules as well as intact, subunit, and peptide levels of antibody analysis. Peak height was also increased using the 1.5 mm i.d. column, although the scale of increase varies between isocratic and gradient modes, likely due to differences in system dispersion effects and variation in electrospray ionization efficiency at different flow rates.