Hydrophilic interaction 10 microm I.D. porous layer open tubular columns for ultratrace glycan analysis by liquid chromatography-mass spectrometry.
Hydrophilic interaction 10 microm I.D. porous layer open tubular columns for ultratrace glycan analysis by liquid chromatography-mass spectrometry.
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DOI：
10.1016/j.chroma.2008.09.105
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发表时间：
2009-02-20
期刊：
影响因子：
--
通讯作者：
Karger BL
中科院分区：
文献类型：
--
作者：
Luo Q;Rejtar T;Wu SL;Karger BL
纳米液相色谱-电喷雾质谱（ESI-MS）分析多糖的灵敏度随流动相流量的减小而增大，同时离子抑制减弱。在本研究中，我们描述了高效10 μ d胺键合聚氯乙烯-二乙烯基苯亲水性相互作用（HILIC）多孔层开管（PLOT）色谱柱的制备和性能，该色谱柱运行速度为20 nL/min，用于多糖混合物的分离和分析。采用原位聚合法制备了具有均匀多孔聚合物层的HILIC-PLOT色谱柱（柱间保留时间RSD ~4%），再用乙二胺一步改性。使用2.5 m × 10 μm i.d的HILIC-PLOT柱与ESI-MS在线耦合时，3-唾液四糖的检出限较低（0.3 fmol）。采用葡聚糖阶梯标准评价色谱柱的性能，在入样量为~50 fmol的情况下，葡聚糖含量可达G22，达到了高效分离。作为色谱柱高灵敏度的一个例子，通过注射10 fmol的中性和唾液化的聚糖，可以对聚糖结构进行MS6表征。作为另一个高灵敏度LC-MS分析3ng卵白蛋白的PNGase F消化的例子，可以从一次分析中自信地鉴定出28个n -链聚糖。获得了每个卵清蛋白聚糖的高质量MS/MS光谱，并手工解释用于结构分析。在超痕量水平上，HILIC PLOT柱是一种非常有前途的LC-MS分析方法。
The sensitivity of glycan analysis using nano-liquid chromatography interfaced with electrospray ionization mass spectrometry (ESI-MS) increases with the decrease of the mobile phase flow rate, accompanied by reduced ion suppression. In this study, we describe the preparation and performance of high efficiency 10 μm i.d. amine-bonded poly(vinylbenzyl chloride-divinylbenzene) hydrophilic interaction (HILIC) porous layer open tubular (PLOT) columns operated at 20 nL/min for the separation and analysis of glycan mixtures. HILIC-PLOT columns with a uniform porous polymer layer were reproducibly prepared (~4% RSD in retention time from column to column) via in-situ polymerization, followed by one step modification with ethylenediamine. When coupled on-line with negative ESI-MS, low detection limits (0.3 fmol) for a 3-sialyl-tetrasaccharide were achieved using a 2.5 m × 10 μm i.d. HILIC-PLOT column. A dextran ladder standard was used to evaluate the performance of the column, and high efficiency separation was achieved with detection of the dextrans up to G22 from ~50 fmol amounts injected. As an example of the high sensitivity of the column, MS6 characterization of glycan structures was possible from the injection of 10 fmol of a neutral and sialylated glycan. As another example of high sensitivity LC-MS analysis of 3 ng of a PNGase F digest of ovalbumin allowed 28 N-linked glycans to be confidently identified from a single analysis. High quality MS/MS spectra for each ovalbumin glycan were acquired and manually interpreted for structure analysis. The HILIC PLOT column is a very promising approach for LC-MS analysis of glycans at the ultratrace level.
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DOI：
10.1016/s0021-9673(00)96972-3
发表时间：
1990-01-19
期刊：
JOURNAL OF CHROMATOGRAPHY
影响因子：
--
作者：
ALPERT, AJ
通讯作者：
ALPERT, AJ
影响因子：
7.4
作者：
GUO, Y;COLON, LA
通讯作者：
COLON, LA
DOI：
10.1016/j.jasms.2005.01.004
发表时间：
2005-05-01
影响因子：
3.2
作者：
Harvey, DJ
通讯作者：
Harvey, DJ
影响因子：
7.4
作者：
Luo, Quanzhou;Yue, Guihua;Karger, Barry L.
通讯作者：
Karger, Barry L.
影响因子：
7.4
作者：
ANDRE, CE;MOSIER, AR
通讯作者：
MOSIER, AR