Analytical and Bioanalytical Chemistry ( IF 3.8 ) Pub Date : 2022-05-26 , DOI: 10.1007/s00216-022-04133-0 Christian Manz 1, 2, 3 , Michael Götze 1, 2 , Clemens Frank 2 , Andreas Zappe 1, 2 , Kevin Pagel 1, 2
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LC–MS is one of the most important tools for the comprehensive characterization of N-glycans. Despite many efforts to speed up glycan analysis via optimized sample preparation (e.g., faster enzyme digestion in combination with instant or rapid labeling dyes), a major bottleneck remains the rather long measurement times of HILIC chromatography. Further complication arises from the necessity to concomitantly calibrate with an external standard to allow for accurate retention times and the conversion into more robust GU values. Here we demonstrate the use of an internal calibration strategy for HILIC chromatography to speed up glycan analysis. By reducing the number of utilized dextran oligosaccharides, the calibrant can be spiked directly into the sample such that external calibration runs are no longer required. The minimized dextran ladder shows accurate GU calibration with a minor deviation of well below 1% and can be applied without modifications in sample preparation or data processing. We further demonstrate the simultaneous use of the minimized dextran ladder as calibrant for the estimation of CCS values in traveling wave ion mobility spectrometry. In both cases, the minimized dextran ladder enables the measurement of calibrant and sample in a single HPLC run without losing information or accuracy.
Graphical abstract
中文翻译：
葡聚糖作为液相色谱与离子淌度质谱联用分析 N-聚糖的内部校准物
LC-MS 是综合表征N的最重要工具之一-聚糖。尽管通过优化样品制备（例如，更快的酶消化结合即时或快速标记染料）来加速聚糖分析，但主要瓶颈仍然是 HILIC 色谱相当长的测量时间。更复杂的情况是需要同时使用外部标准进行校准，以实现准确的保留时间并转换为更可靠的 GU 值。在这里，我们展示了使用 HILIC 色谱的内部校准策略来加速聚糖分析。通过减少使用的葡聚糖寡糖的数量，校准物可以直接加标到样品中，从而不再需要外部校准运行。最小化的葡聚糖阶梯显示准确的 GU 校准，微小偏差远低于 1%，无需修改样品制备或数据处理即可应用。我们进一步证明了同时使用最小化的葡聚糖阶梯作为校准物来估计行波离子迁移谱中的 CCS 值。在这两种情况下，最小化的葡聚糖阶梯可以在单次 HPLC 运行中测量校准物和样品，而不会丢失信息或准确性。
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