Name: an efficient sample preparation method to enhance carbohydrate ion signals in matrix-assisted laser desorption/ionization mass spectrometry
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Description: Watch this Scientific Journal Video about An Efficient Sample Preparation Method to Enhance Carbohydrate Ion Signals in Matrix-assisted Laser Desorption/Ionization Mass Spectrometry at JoVE.com

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1. 样品板预适应
<ol><li>
样品板的清洗
	<ol>
<li>在清洗过程中, 应佩戴丁腈橡胶手套, 以避免样品板的污染。</li>
		<li>用100.0 毫升洗涤剂溶液 (1.0 毫克/毫升) 洗手样品板。</li>
		<li>用蒸馏-去离子水 (DDW) 洗手样品板。</li>
		<li>用30.0 毫升的甲醇冲洗样品板表面。</li>
		<li>把样品板放在600毫升烧杯中, 装满 DDW, 直到样品板完全浸入水中。</li>
		<li>把烧杯放在超声波浴中 (见 <stron...

<ol>
	<li>Holme, D. J., Peck, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Analytical Biochemistry. , (1998).</li><li>Costello, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time,+life+...+and+mass+spectrometry+-+New+techniques+to+address+biological+questions.">Time, life ... and mass spectrometry - New techniques to address biological questions.</a> Biophysical Chemistry. 68 (1-3), 173-188 (1997).</li><li>Caroff, M., Karibian, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+of+bacterial+lipopolysaccharides.">Structure of bacterial lipopolysaccharides.</a> Carbohydrate Research. 338 (23), 2431-2447 (2003).</li><li>Marvin, L. F., Roberts, M. A., Fay, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix-assisted+laser+desorption/ionization+time-of-flight+mass+spectrometry+in+clinical+chemistry.">Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in clinical chemistry.</a> Clinica Chimica Acta. 337 (1), 11-21 (2003).</li><li>Harvey, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Matrix-assisted+laser+desorption/ionization+mass+spectrometry+of+carbohydrates.">Matrix-assisted laser desorption/ionization mass spectrometry of carbohydrates.</a> Mass Spectrometry Reviews. 18 (6), 349-450 (1999).</li><li>Ciucanu, I., Kerek, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+and+rapid+method+for+the+permethylation+of+carbohydrates.">A simple and rapid method for the permethylation of carbohydrates.</a> Carbohydrate Research. 131 (2), 209-217 (1984).</li><li>Lamari, F. N., Kuhn, R., Karamanos, N. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Derivatization+of+carbohydrates+for+chromatographic,+electrophoretic+and+mass+spectrometric+structure+analysis.">Derivatization of carbohydrates for chromatographic, electrophoretic and mass spectrometric structure analysis.</a> Journal of Chromatography B. 793 (1), 15-36 (2003).</li><li>Nishikaze, T., Amano, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reverse+thin+layer+method+for+enhanced+ion+yield+of+oligosaccharides+in+matrix-assisted+laser+desorption/ionization.">Reverse thin layer method for enhanced ion yield of oligosaccharides in matrix-assisted laser desorption/ionization.</a> Rapid Communications in Mass Spectrometry. 23 (23), 3787-3794 (2009).</li><li>Williams, T. I., Saggese, D. A., Wilcox, R. J., Martin, J. D., Muddiman, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+matrix+crystal+structure+on+ion+abundance+of+carbohydrates+by+matrix-assisted+laser+desorption/ionization+Fourier+transform+ion+cyclotron+resonance+mass+spectrometry.">Effect of matrix crystal structure on ion abundance of carbohydrates by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry.</a> Rapid Communications in Mass Spectrometry. 21 (5), 807-811 (2007).</li><li>Nicola, A. J., Gusev, A. I., Proctor, A., Jackson, E. K., Hercules, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+the+fast-evaporation+sample+preparation+method+for+improving+quantification+of+angiotensin+II+by+matrix-assisted+laser+desorption/ionization.">Application of the fast-evaporation sample preparation method for improving quantification of angiotensin II by matrix-assisted laser desorption/ionization.</a> Rapid Communications in Mass Spectrometry. 9 (12), 1164-1171 (1995).</li><li>Lai, Y. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reducing+Spatial+Heterogeneity+of+MALDI+Samples+with+Marangoni+Flows+During+Sample+Preparation.">Reducing Spatial Heterogeneity of MALDI Samples with Marangoni Flows During Sample Preparation.</a> Journal of the American Society for Mass Spectrometry. 27 (8), 1314-1321 (2016).</li><li>Ou, Y. -. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+Homogeneous+MALDI+Samples+for+Quantitative+Applications.">Preparation of Homogeneous MALDI Samples for Quantitative Applications.</a> Journal of Visualized Experiments. (116), e54409 (2016).</li><li>Lee, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancing+carbohydrate+ion+yield+by+controlling+crystalline+structures+in+matrix-assisted+laser+desorption/ionization+mass+spectrometry.">Enhancing carbohydrate ion yield by controlling crystalline structures in matrix-assisted laser desorption/ionization mass spectrometry.</a> Analytica Chimica Acta. , 49-55 (2017).</li><li>Allwood, D. A., Perera, I. K., Perkins, J., Dyer, P. E., Oldershaw, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+'near'+homogeneous+samples+for+the+analysis+of+matrix-assisted+laser+desorption/ionisation+processes.">Preparation of 'near' homogeneous samples for the analysis of matrix-assisted laser desorption/ionisation processes.</a> Applied Surface Science. 103 (3), 231-244 (1996).</li><li>Sadeghi, M., Vertes, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crystallite+size+dependence+of+volatilization+in+matrix-assisted+laser+desorption+ionization.">Crystallite size dependence of volatilization in matrix-assisted laser desorption ionization.</a> Applied Surface Science. 127 (Supplement C), 226-234 (1998).</li></ol>

样本异质性是 MALDI 中最常用的样品制备方法, 但合成的晶体是高度异构的。此类样本显示了低投篮和采样采样信号重现性。因此, 在数据采集过程中寻找样本区域中的 "甜点" 是 MALDI 实验中的一个常见过程。这种非均匀样本在常规分析中不适合量化。
在目前的研究中, MALDI 样品形态学通过再结晶优化。再结晶对碳水化合物信号强度和数据稳定性的改善归因于碳水化合物与基质?...

碳水化合物分析是一个重要而具有挑战性的课题。碳水化合物和他们的衍生物在活有机体扮演重要角色1,2,3。这些分子结构复杂, 易于分解。由于分离和检测方面的困难, 其中许多不能清楚地描绘出来。虽然基质辅助激光解吸/电离 (MALDI) 质谱 (MS) 已被应用于分析范围广泛的生物分子, 由于其灵敏度和可理解的结果4, 分析碳水化合物使用 MALDI-MS 继续由于这种分子的电离效率低5, 这是一个主要的挑战。化学衍生是提高碳水化合物6、7的电离效率的常用方法, 但这种过程是时间和样本消耗。此外, 衍生物碳水化合物的电离效率仍然低于蛋白质。因此, 在不需要复杂程序的情况下, 改进 MALDI 中碳水化合物信号的方法是必要的。
MALDI-MS 在定量分析中的应用是另一个具有挑战性的课题。MALDI 的一个主要问题是它的灵敏度和数据重现性主要依赖于样品制备协议和实验参数。在许多情况下, 由于异质样品形貌和分析物分布, MALDI 的定量分析是不可靠的。一个众所周知的例子是用 25-苯甲酸酸 (DHB) MALDI 基质制备的样品。当 DHB 在周围环境下慢慢结晶时, 分析物被纳入基体晶体的程度是不可预知的, 因为结果样品显示不规则的形貌。这种样品通常由大针状和细晶体组成。当 DHB 使用挥发性溶剂和/或加热样品板制备时, 快速干燥过程会产生更均匀的细晶, 并取得更好的定量结果8、9、10。这种技术被称为 MALDI 样品的 "再结晶"。这种改进归因于在快速结晶过程中更好地将分析物纳入精细基体晶体中。我们还表明, 调整样品制备环境减少了碳水化合物信号的异质性和改进的定量结果11,12。这些研究结果表明, 样本形态学是确定碳水化合物信号质量的关键因素。为了制定日常分析的一般策略, 需要有一个有效的样本改革方法来提高碳水化合物的灵敏度。
我们有系统地研究了 MALDI-MS 样品形态学和碳水化合物敏感性的相关性, 在最近的报告13。利用几个重要的碳水化合物和矩阵得到的结果表明, 重结晶干 MALDI 样品能较好地实现最佳信号增强。用甲醇 (甲醇) 快速再结晶法对用常规干液滴 (DD) 方法制备的试样形貌进行了改造。详细的样品准备协议在这里展示。该协议包括样品板预处理、样品沉积和再结晶、质谱分析等三主要步骤。所使用的碳水化合物包括唾液酸化酶 (SLea) 和 maltoheptaose (MH)。DHB 用作模型矩阵。结果表明, 再结晶后碳水化合物信号强度和空间分布明显改善。这种方法可应用于其他常用基质的样品, 包括24、6-trihydroxyacetophenone (行动计划推行) 和αcyano-4-hydroxycinnamic 酸。这种方法是一种一般的方法, 可以很容易地集成在实验室例行的碳水化合物分析。

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			<td height="23" style="height:23px;width:332px;">Reagent</td>
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			<td style="width:137px;"></td>
			<td style="width:191px;"></td>
		</tr>
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			<td height="23" style="height:23px;width:332px;">Detergent powder</td>
			<td style="width:157px;">Alconox</td>
			<td style="width:137px;">242985</td>
			<td></td>
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			<td style="width:157px;">Merck</td>
			<td style="width:137px;">106009</td>
			<td></td>
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			<td height="23" style="height:23px;width:332px;">Acetonitrile</td>
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			<td style="width:137px;">100003</td>
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			<td height="23" style="height:23px;">2,5-dihydroxybenzoic acid (DHB)</td>
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			<td style="width:137px;">A11459</td>
			<td></td>
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			<td style="width:137px;">S1782</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Maltoheptaose</td>
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			<td style="width:137px;">M7753</td>
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			<td height="23" style="height:23px;width:332px;">600 mL beaker</td>
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			<td style="width:137px;">2110648</td>
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			<td height="23" style="height:23px;width:332px;">Hygrometer</td>
			<td style="width:157px;">Wisewind</td>
			<td style="width:137px;">5330</td>
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		<tr height="23">
			<td height="23" style="height:23px;width:332px;">Nitrogen gas flowmeter</td>
			<td style="width:157px;">Dwyer</td>
			<td style="width:137px;">RMA-6-SSV</td>
			<td></td>
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		<tr height="23">
			<td height="23" style="height:23px;width:332px;">K-type thermocouples</td>
			<td style="width:157px;">Digitron</td>
			<td style="width:137px;">311-1670</td>
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			<td style="width:137px;">&#160;SI-0236</td>
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			<td height="23" style="height:23px;width:332px;">Mini centrifuge</td>
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			<td height="23" style="height:23px;width:332px;">Temperature controllable drying chamber</td>
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			<td></td>
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			<td height="23" style="height:23px;width:332px;">Ultraflex II TOF/TOF mass spectrometer</td>
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			<td style="width:137px;"></td>
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			<td height="23" style="height:23px;width:332px;">MTP 384 target plate polished steel BC</td>
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			<td style="width:137px;">8280781</td>
			<td></td>
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		<tr height="23">
			<td height="23" style="height:23px;width:332px;">Flexcontrol Version 3.4</td>
			<td style="width:157px;">Bruker Daltonics</td>
			<td style="width:137px;"></td>
			<td>Control software</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:332px;">Fleximaging Version 2.1</td>
			<td style="width:157px;">Bruker Daltonics</td>
			<td style="width:137px;"></td>
			<td>Imaging software</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:332px;">Flexanalysis Version 3.4</td>
			<td style="width:157px;">Bruker Daltonics</td>
			<td style="width:137px;"></td>
			<td>Analysis software</td>
		</tr>
	</tbody>
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an efficient sample preparation method to enhance carbohydrate ion signals in matrix-assisted laser desorption/ionization mass spectrometry

样品制备是质谱 (MS) 分析碳水化合物的关键过程。虽然基质辅助激光解吸/电离 (MALDI) MS 是碳水化合物分析中的选择方法, 但碳水化合物样品的不良离子信号和数据重现性仍然是严重的问题。对于碳水化合物的定量分析, 必须有一个有效的分析协议, 提供卓越的数据质量。该视频演示了用于提高信号强度和减少 MALDI 中碳水化合物数据变化的样本准备协议。在样品滴的干燥和结晶后, 在质谱分析前, 甲醇对晶体形态进行了改造。用 MALDI 成像质谱 (IMS) 检测碳水化合物信号的增强。实验结果表明, 晶体的改造调整了晶体结构, 重新分配了碳水化合物的分析。与常规 MALDI 中的干液滴制备方法相比, 甲醇转化碳水化合物晶体形貌明显改善了信号强度、离子图像分布和数据稳定性。由于此处演示的协议不涉及样本组成的变化, 它们通常适用于各种碳水化合物和矩阵。

图 1显示了用 DD 和再结晶方法制备的 SLeA混合 DHB 的代表性 SEM 图像。DD 法制备的典型 DHB 形态为边缘处的大针状晶体和样品点中心的精细晶体结构。这种针状晶体的典型长度为100µm。经甲醇再结晶后, 试样的面积较大, 均匀地覆盖着细小的鳞片状晶体。"鳞片" 晶体的长度约为20-50 µm. 再结晶样品提供比传统 DD 样品所生产的更大的有效表面积。</...

Watch this Scientific Journal Video about An Efficient Sample Preparation Method to Enhance Carbohydrate Ion Signals in Matrix-assisted Laser Desorption/Ionization Mass Spectrometry at JoVE.com

An Efficient Sample Preparation Method to Enhance Carbohydrate Ion Signals in Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

本文演示了在样品制备过程中, 通过改造晶体结构来增强 MALDI 质谱中的碳水化合物离子信号的协议。

基质辅助激光解吸/电离质谱中增强碳水化合物离子信号的高效样品制备方法

Sample preparation is a critical process in mass spectrometry (MS) analysis of carbohydrates. Although matrix-assisted laser desorption/ionization (MALDI) ...

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Sequencing of Plant Wall Heteroxylans Using Enzymic, Chemical Methylation and Physical Mass Spectrometry, Nuclear Magnetic Resonance Techniques

使用酶法，化学（甲基化）和物理（质谱，核磁共振）技术，植物墙Heteroxylans测序

This protocol describes the specific techniques used for the characterization of reducing end (RE) and internal region glycosyl sequence(s) of ...

An HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry

液相色谱离子迁移 QTOF 质谱测定蛋白质生物中宿主细胞蛋白的 HS-MRM 法

The analysis of low-level (1-100 ppm) protein impurities (e.g., host-cell proteins (HCPs)) in protein biotherapeutics is a challenging assay requiring ...

Characterization of Synthetic Polymers via Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) Mass Spectrometry

Characterization of Synthetic Polymers via Matrix Assisted Laser Desorption Ionization Time of Flight MALDI-TOF Mass Spectrometry

基质辅助激光解吸电离飞行时间 (MALDI) 质谱法表征合成聚合物

There are many techniques that can be employed in the characterization of synthetic homopolymers, but few provide as useful of information for end group ...

Imaging Corrosion at the Metal-Paint Interface Using Time-of-Flight Secondary Ion Mass Spectrometry

利用飞行时二次离子质谱法成像金属颜料界面的腐蚀

Corrosion developed at the paint and aluminum (Al) metal-paint interface of an aluminum alloy is analyzed using time-of-flight secondary ion mass ...

Ion Mobility-Mass Spectrometry Techniques for Determining the Structure and Mechanisms of Metal Ion Recognition and Redox Activity of Metal Binding Oligopeptides

用于测定金属结合寡聚肽的金属电子识别和机制的Ion流动性-质谱技术

Electrospray ionization (ESI) can transfer an aqueous-phase peptide or peptide complex to the gas-phase while conserving its mass, overall charge, ...

Analysis of Complex Molecules and Their Reactions on Surfaces by Means of Cluster-Induced Desorption/Ionization Mass Spectrometry

聚类诱导解吸/电电质谱法对复杂分子及其在表面的反应分析

Desorption/Ionization Induced by Neutral SO2 Clusters (DINeC) is employed as a very soft and efficient desorption/ionization technique for mass ...

3D Depth Profile Reconstruction of Segregated Impurities Using Secondary Ion Mass Spectrometry

使用二次离子质谱法重建分离杂质的 3D 深度剖面

The presented protocol combines excellent detection limits (1 ppm to 1 ppb) using secondary ion mass spectrometry (SIMS) with reasonable spatial ...