C60-based chemical labeling strategy for the determination of polyamines in biological samples using matrix-assisted laser desorption/ionization mass spectrometry
C60-based chemical labeling strategy for the determination of polyamines in biological samples using matrix-assisted laser desorption/ionization mass spectrometry
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基于 C60 的化学标记策略，使用基质辅助激光解吸/电离质谱法测定生物样品中的多胺
DOI：
10.1016/j.talanta.2020.121790
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发表时间：
2021
期刊：
影响因子：
6.1
通讯作者：
Feng Yu-Qi
中科院分区：
文献类型：
--
作者：
Qin Zhang-Na;Yu Qiong-Wei;Zhou Ping;Feng Yu-Qi
生物活性多胺在基因表达、细胞生长、蛋白质合成和信号转导等生物过程中发挥着重要作用。准确测定多胺有助于研究其生物学功能。为此，提出了一种基于C60的化学标记策略，用于测定生物样品中的多胺(腐胺、身体胺、精胺和精胺)，并建立了基质辅助激光解吸电离质谱仪(MALDI MS)。用N-羟基琥珀酰亚胺酯功能化的C60(NHS-C60)作为标记剂，标记的多胺的Tem/Z高达900 kDa以上，避免了低m/z区的基质干扰。此外，作为NHS-C60的衍生物，还同时产生了单取代和双取代的多胺，这有利于多胺的定性分析。用NHS-C60标记的细胞和小鼠粪便样品中的多胺对分析方法进行验证。线性关系良好，相关系数在0.9786~0.9982之间。定量限在0.68~1.48 mg/L之间。方法的重现性和可靠性良好，日内和日间精密度为2.8%~16.6%，回收率为81.8%~119.9%。最后，将该方法应用于细胞和小鼠粪便中多胺的测定。在细胞内检测到三种多胺，高脂饮食小鼠粪便中身体胺和亚精胺的含量显著低于正常饮食小鼠。结果表明，NHS-C60标记与MALDI-MS联用技术适用于生物样品中多胺类化合物的测定。
Bioactive polyamines play important roles in many biological processes such as gene expression, cell growth, protein synthesis, and signal transduction. Accurate determination of polyamines is helpful for studying their biological functions. Herein, a C60-based chemical labeling strategy was proposed for the determination of polyamines (putrescine, cadaverine, spermidine, and spermine) in biological samples using matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). An N-hydroxysuccinimide ester functionalized C60(NHS–C60) was used as a labeling reagent and them/zof the labeled polyamines reached up to more than 900 Da, which avoided matrix interferences in the lowm/zregion. In addition, as NHS–C60derivatives, mono- and bis-substituted polyamines were produced simultaneously, which benefited the qualitative analysis of polyamines. The analytical method was validated using NHS–C60labeled polyamines in cells and mice feces samples. Good linearities were obtained with correlation coefficients ranging from 0.9786 to 0.9982. The limits of quantification were in the range of 0.68–1.48 pmol. Good reproducibility and reliability of our proposed method were confirmed by intra- and inter-day precisions ranged from 2.8 to 16.6%, and the recoveries ranged between 81.8 and 119.9%. Finally, the proposed method was applied to determine polyamines in cells and mice feces. Three polyamines were detected in the cells, and the contents of cadaverine and spermidine in the feces of high-fat diet mice were found to be significantly lower than those in the normal diet mice. The results show that the proposed NHS–C60labeling coupled with MALDI MS strategy is suitable for the determination of polyamines in biological samples.