Development of C60-based labeling reagents for the determination of low -molecular-weight compounds by matrix assisted laser desorption ionization mass (I): Determination of amino acids in microliter biofluids
Development of C60-based labeling reagents for the determination of low -molecular-weight compounds by matrix assisted laser desorption ionization mass (I): Determination of amino acids in microliter biofluids
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开发基于 C60 的标记试剂，用于通过基质辅助激光解吸电离质量测定低分子量化合物（I）：微升生物流体中氨基酸的测定
DOI：
10.1016/j.aca.2017.01.018
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发表时间：
2017
影响因子：
6.2
通讯作者：
Feng Yu-Qi
中科院分区：
文献类型：
--
作者：
Wu Pin;Xiao Hua-Ming;Jun Ding;Deng Qian-Yun;Zheng Fang;Feng Yu-Qi
由于基质辅助激光解吸/电离飞行时间质谱仪(MALDI-MS)在低m/z区存在基质信号干扰，且MS响应的重现性较差，因此使用基质辅助激光解吸/电离飞行时间质谱仪(MALDI-MS)对低分子化合物(<；800×Da)进行定量是一项具有挑战性的工作。本研究建立了一种快速、灵敏、可靠的C60标记-MALDI-MS方法测定生物体液中的氨基酸。合成了N-羟基琥珀酰亚胺功能化的C60作为标记剂，并将其作为880Da的标记添加到AAs中；以含有C60的羧酸作为内标对MS变异进行归一化。这解决了MALDI-MS用于小分子分析的固有问题。整个分析过程-包括简单的蛋白质沉淀和在MALDI MS分析之前在微波中10分钟的衍生化-可以在20分钟内完成，吞吐量高，样品基质容差大。AA定量在0.9954.7~70.0μM范围内呈良好的线性关系，相关系数(R)>0.0 5。方法的检出限为70-300.0毫微克分子。方法的重现性和可靠性良好，日内和日间精密度RSD均小于13.8%，体液中回收率在80.4%~106.8%之间。该方法可用于尿液、血清、血浆、全血和脑脊液的1次μL。最重要的是，C60标记策略是MALDI MS分析各种低分子化合物的通用方法，因为功能化的C60现在是按需提供的。
Quantification of low molecular weight compounds (<800 Da) using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI MS) is challenging due to the matrix signal interference at lowm/zregion and poor reproducibility of MS responses. In this study, a C60 labeling-MALDI MS strategy was proposed for the fast, sensitive and reliable determination of amino acids (AAs) in biofluids. AnN-hydroxysuccinimide functionalized C60 was synthesized as the labeling reagent and added as an 880 Da tag to AAs; a carboxyl acid containing C60 was employed as the internal standards to normalize MS variations. This solved the inherent problems of MALDI MS for small molecule analysis. The entire analytical procedure—which consisted of simple protein precipitation and 10 min of derivatization in a microwave prior to the MALDI MS analysis—could be accomplished within 20 min with high throughput and great sample matrix tolerance. AA quantification showed good linearity from 0.7 to 70.0 μM with correlation coefficients (R) larger than 0.9954. The limits of detection were 70.0–300.0 fmol. Good reproducibility and reliability of the method were demonstrated by intra-day and inter-day precision with relative standard deviations less than 13.8%, and the recovery in biofluid ranged from 80.4% to 106.8%. This approach could be used in 1 μL of urine, serum, plasma, whole blood, and cerebrospinal fluid. Most importantly, the C60 labeling strategy is a universal approach for MALDI MS analysis of various LMW compounds because functionalized C60 is now available on demand.