Determination of ricin by nano liquid chromatography/mass spectrometry after extraction using lactose-immobilized monolithic silica spin column
Determination of ricin by nano liquid chromatography/mass spectrometry after extraction using lactose-immobilized monolithic silica spin column
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DOI：
10.1002/jms.1953
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发表时间：
2011-08-01
影响因子：
2.3
通讯作者：
Seto, Yasuo
中科院分区：
文献类型：
--
作者：
Kanamori-Kataoka, Mieko;Kato, Haruhito;Seto, Yasuo
蓖麻毒素是一种糖化蛋白毒素，已被《化学武器公约》列为有毒物质。目前的检测方法可能导致假阴性和/或阳性，其标准不是基于蛋白质氨基酸序列的鉴定。本研究采用乳糖固定化硅胶整体萃取-胰蛋白酶消化-液相色谱/质谱联用（LC/MS）技术快速、准确地测定蓖麻毒素。将固定在单片二氧化硅上的乳糖作为捕获配体从样品溶液中提取蓖麻毒素，并将二氧化硅支撑在圆盘填充的自旋柱中。蓖麻毒素回收率达40%以上。提取后，胰酶消化，液相色谱质谱分析。分别用傅里叶变换-质谱和线性离子阱-质谱测定分离肽峰分子离子的准确质量和质谱。选择从蓖麻毒素A链（m/z 537.8、448.8和586.8）和b链（m/z 701.3、647.8和616.8）中分离得到的6条多肽作为鉴定蓖麻毒素的标记肽。在这些标记肽中，有两个肽是蓖麻毒素特异性的。本方法适用于粗样品中蓖麻毒素的测定。单片硅胶萃取去除了样品总离子色谱中的大部分污染物峰，并通过LC/MS清晰地检测到6种标记肽。整个处理过程对蓖麻毒素8 ng/ml以上的检测鉴定耗时约5小时，完全可以处理使用化学武器的恐怖主义。版权所有：John Wiley & Sons， Ltd。
Ricin is a glycosylated proteinous toxin that is registered as toxic substance by Chemical Weapons convention. Current detection methods can result in false negatives and/or positives, and their criteria are not based on the identification of the protein amino acid sequences. In this study, lactose-immobilized monolithic silica extraction followed by tryptic digestion and liquid chromatography/mass spectrometry (LC/MS) was developed as a method for rapid and accurate determination of ricin. Lactose, which was immobilized on monolithic silica, was used as a capture ligand for ricin extraction from the sample solution, and the silica was supported in a disk-packed spin column. Recovery of ricin was more than 40%. After extraction, the extract was digested with trypsin and analyzed by LC/MS. The accurate masses of molecular ions and MS/MS spectra of the separated peptide peaks were measured by Fourier transform-MS and linear iontrap-MS, respectively. Six peptides, which were derived from the ricin A-(m/z 537.8, 448.8 and 586.8) and B-chains (m/z 701.3, 647.8 and 616.8), were chosen as marker peptides for the identification of ricin. Among these marker peptides, two peptides were ricin-specific. This method was applied to the determination of ricin from crude samples. The monolithic silica extraction removed most contaminant peaks from the total ion chromatogram of the sample, and the six marker peptides were clearly detected by LC/MS. It takes about 5 h for detection and identification of more than 8 ng/ml of ricin through the whole handling, and this procedure will be able to deal with the terrorism using chemical weapon. Copyright (C) 2011 John Wiley & Sons, Ltd.