<br>2,5-二羟基对苯二甲酸：用于改进氨基酸检测和成像的基质,Analytical Chemistry

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2,5-二羟基对苯二甲酸：用于改进氨基酸检测和成像的基质
Analytical Chemistry ( IF 6.7 ) Pub Date : 2023-11-28 , DOI: 10.1021/acs.analchem.3c01731
Jinxiang Fu _{1,

2} , Jianchi Gu _{1,

2} , Zhibin Bao _{1,

2} , Yunpeng Zhou ₃ , Hao Hu _{1,

2} , Chenyu Yang _{1,

2} , Ran Wu _{1,

2} , Haiqiang Liu _{1,

2} , Liang Qin _{1,

2} , Hualei Xu _{1,

2} , Jinrong Li _{1,

2} , Hua Guo _{1,

2} , Lei Wang _{1,

2} , Yijun Zhou ₂ , Xiaodong Wang _{1,

2} , Gaopeng Li ₃

Affiliation

氨基酸 (AA) 是低分子量 (low-MW) 代谢物，不仅是蛋白质合成的重要组成部分，也是维持生命系统中氮平衡的重要组成部分。 AA 的原位检测和成像对于理解更复杂的生物过程至关重要。基质辅助激光解吸/电离质谱成像（MALDI-MSI）是一种无标记质谱成像技术，能够同时检测和成像生物样品中不同内源/外源化合物的空间分布和相对丰度。 MALDI-MSI的优异效率归功于MALDI矩阵的选择。然而，据我们所知，还没有专门为 AA 开发的矩阵。在此，我们报告了一种 MALDI 基质，2,5-二羟基对苯二甲酸 (DHT)，它可以改善 MALDI-MS 对生物样品中 AA 的检测和成像。我们的结果表明，DHT 表现出强紫外可见光 (UV-vis) 吸收、均匀的基质沉积和高真空稳定性。此外，与常用的 2,5-二羟基苯甲酸 (DHB) 和 α-氰基-4-羟基肉桂酸基质相比，DHT 产生的基质相关离子信号在m / z < 500 时分别减少了 50% 和 71.8%（ CHCA)，分别在各自的有机溶剂中。在定量性能方面，使用 DHT 作为基质，可以检测精氨酸、谷氨酸、谷氨酰胺和脯氨酸，检测限分别为 6、4、6 和 4 ng/mL。使用 DHT 作为基质，MALDI-MS 成功检测到人血清中的所有 20 种蛋白质 AA，而使用 DHB 和 CHCA 基质时分别仅检测到 7 种和 10 种 AA。此外，首次在可食用巨牡蛎（牡蛎）组织中成功检测到 20 种蛋白质 AA 和牛磺酸并对其进行成像。我们的研究表明，使用 DHT 作为基质可以改善 MALDI-MS 对生物样品中 AA 的检测和成像。

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2,5-Dihydroxyterephthalic Acid: A Matrix for Improved Detection and Imaging of Amino Acids

Amino acids (AAs), which are low-molecular-weight (low-MW) metabolites, serve as essential building blocks not only for protein synthesis but also for maintaining the nitrogen balance in living systems. In situ detection and imaging of AAs are crucial for understanding more complex biological processes. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) is a label-free mass spectrometric imaging technique that enables the simultaneous detection and imaging of the spatial distribution and relative abundance of different endogenous/exogenous compounds in biological samples. The excellent efficiency of MALDI-MSI is attributed to the choice of the MALDI matrix. However, to the best of our knowledge, no matrix has been specifically developed for AAs. Herein, we report a MALDI matrix, 2,5-dihydroxyterephthalic acid (DHT), which can improve the detection and imaging of AAs in biological samples by MALDI-MS. Our results indicated that DHT exhibited strong ultraviolet–visible (UV–vis) absorption, uniform matrix deposition, and high vacuum stability. Moreover, the matrix-related ion signals produced from DHT were reduced by 50 and 71.8% at m/z < 500 compared to the commonly used matrices of 2,5-dihydroxybenzoic acid (DHB) and α-cyano-4-hydroxycinnamic acid (CHCA), respectively, in their respective organic solvents. In terms of quantitative performance, arginine, glutamic acid, glutamine, and proline can be detected with limits of detection of 6, 4, 6, and 4 ng/mL, respectively, using the DHT as the matrix. Using DHT as the matrix, all 20 protein AAs were successfully detected in human serum by MALDI-MS, whereas only 7 and 10 AAs were detected when DHB and CHCA matrices were used, respectively. Furthermore, 20 protein AAs and taurine were successfully detected and imaged in a section of edible Crassostrea gigas (oyster) tissue for the first time. Our study demonstrates that using DHT as a matrix can improve the detection and imaging of AAs in biological samples by MALDI-MS.

更新日期：2023-11-28

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