Development of C60-based labeling reagents for the determination of low-molecular-weight compounds by matrix assisted laser desorption ionization mass spectrometry (II): Determination of thiols in human serum
Development of C60-based labeling reagents for the determination of low-molecular-weight compounds by matrix assisted laser desorption ionization mass spectrometry (II): Determination of thiols in human serum
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基质辅助激光解吸电离质谱测定低分子化合物的C60标记试剂的研制（二）：人血清中硫醇的测定
DOI：
10.1016/j.aca.2020.01.027
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发表时间：
2020
影响因子：
6.2
通讯作者：
Feng Yu-Qi
中科院分区：
文献类型：
--
作者：
Qin Zhang-Na;Ding Jun;Yu Qiong-Wei;Qi Chu-Bo;Wu Dong-Mei;Zhou Ping;Feng Yu-Qi
生物体液中硫醇稳态的扰动被认为与几种疾病有关，因此需要用于测定人血浆或血清中低分子量（LMW）硫醇的可靠分析方法。建立了基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）法测定人血清中谷胱甘肽、半胱氨酸、同型半胱氨酸和半胱氨酰甘氨酸4种低分子量巯基化合物的方法。它基于使用溴乙酰基官能化的C60（Br-C60）作为标记硫醇的衍生化试剂。Br-C60标记可以将832-Da标签添加到硫醇，这将硫醇信号移动到高质量区域，有效避免了传统MALDI基质在800 Da以下产生的信号干扰。在微波辅助条件下，标记可在5 min内完成。因此，基于Br-C60标记的MALDI-TOF MS分析方法可以实现血清中低分子量硫醇的高通量分析。在0.5-500.0 μM范围内，人血清中硫醇的线性关系良好，相关系数（R）大于0.9960。方法的检出限为0.07-0.18 μM，相对标准偏差小于13.5%，回收率为81.9 ~ 117.1%。用该法测定了乳腺癌患者血清中的四种巯基。结果显示，BC血清样本中四种硫醇的含量与健康对照（HC）相比发生了显着变化。
Perturbation of thiol homeostasis in biological fluids are thought to be associated with several diseases, and reliable analytical methods for the determination of low molecular weight (LMW) thiols in human plasma or serum are thus required. In this study, a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method is described for high throughput determination of four LMW thiols (glutathione, cysteine, homocysteine and cysteinylglycine) in human serum. It is based on the use of a bromoacetyl functionalized C60 (Br–C60) as a derivatization reagent to label thiols. The Br–C60 labeling can add an 832-Da tag to thiols, which moves thiol signals to high mass region and effectively avoids the signal interference generated by the traditional MALDI matrix below 800 Da. The labeling can be completed within 5 min under microwave-assisted condition. Thereby, the Br–C60 labeling based MALDI-TOF MS analytical method can achieve high throughput analysis of LMW thiols in serum. Good linearities of the method for the thiols in human serum were obtained in the range of 0.5–500.0 μM with correlation coefficient (R) greater than 0.9960. The limit of detection is in the range of 0.07–0.18 μM for the investigated thiols in human serum with relative standard deviations of lower than 13.5% and recoveries ranging from 81.9 to 117.1%. Using the method, four thiols in microliter serum samples of breast cancer (BC) patients were determined. The result showed that the contents of the four thiols in BC serum samples significantly changed compared to the healthy control (HC).