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Novel Nanomaterials-Based Fluorescent Assay for Rapid Detection of Glutathione
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摘要: 血清中谷胱甘肽（GSH）的含量水平与多种疾病的发生相关，本文成功建立了一种能快速灵敏测量GSH含量的荧光法。以二氧化硅纳米颗粒为载体和稳定剂，在超声条件下简单、快速地合成了二氧化锰包覆的二氧化硅纳米颗粒（SiO2@MnO2）；以精氨酸和胸腺嘧啶为配体合成了具有高荧光量子产率的荧光金纳米簇（Arg/ATT/AuNCs）。实验结果表明GSH的强还原性可以分解MnO2释放出Mn(II)猝灭Arg/ATT/AuNCs的荧光，因此建立了基于SiO2@MnO2和Arg/ATT/AuNCs的荧光法测量血清中GSH。在最优条件下，该方法定量测量GSH的线性范围为2.5～20 nmol/L，最低检测限为1.23 nmol/L，优于已报道的类似方法。本方法测量低、中、高浓度GSH的相对扩展不确定度分别为8.6%、12.4%、18.8%（k = 2），准确度在87.0%～105.6%之间，满足GSH检测的需求，为血清中GSH含量的灵敏、准确测量提供了新技术。Abstract: The level of glutathione (GSH) in serum has been correlated with the occurrence of various diseases. In this paper, a fluorescence method that can measure GSH levels rapidly and sensitively was successfully established. In this method, manganese dioxide-coated silica nanoparticles (SiO2@MnO2) were synthesized simply and rapidly under ultrasonic conditions using silica nanoparticles as carriers and stabilizers, and fluorescent gold nanoclusters (Arg/ATT/AuNCs) with high fluorescence quantum yields were synthesized using arginine and thymine as ligands. The experimental results showed that the strong reduction of GSH could decompose MnO2 to release Mn(II) to burst the fluorescence of Arg/ATT/AuNCs, so a fluorescence method based on SiO2@MnO2 and Arg/ATT/AuNCs was established to measure GSH in serum. Under optimal conditions, the linear range of the method for quantitative measurement of GSH was from 2.5 to 20 nmol/L with a minimum detection limit of 1.23 nmol/L, which was better than similar methods that have been reported. The relative extended uncertainties of the method were 8.6%, 12.4%, and 18.8% (k = 2) for low, medium, and high GSH concentrations, respectively, with accuracies ranging from 87.0% to 105.6%, which meet the requirements of GSH determination and provide a new technique for sensitive and accurate measurement of GSH in serum.
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Key words:
- glutathione /
- manganese dioxide /
- fluorescent gold nanoclusters /
- fluorescence method /
- rapid detection /
- nanomaterials
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图 1 基于新型纳米材料的荧光法快速检测GSH原理示意图
Figure 1. Scheme of the novel nanomaterial-based fluorescent assay for GSH rapid detection
图 4 GSH、SiO2@MnO2以及两者的混合液加入到Arg/ATT/AuNCs中的荧光发射光谱图，内嵌图为在紫外灯下四种溶液体系的荧光成像
Figure 4. The spectrum of Arg/ATT/AuNCs (green) with GSH (blue), SiO2@MnO2(red) and both of them (black). The inset shows the fluorescence imaging of the four solution under UV-Vis
图 6 血清样品中不同含量GSH的检测
Figure 6. Detection of GSH with different concentration in serum samples
表 1 血清样品中GSH的测量准确度和精密度（n=3）
Table 1. The accuracy and precision of GSH measurement in serum samples (n=3)
样品 检测浓度
(nmol/L)添加浓度
(nmol/L)平均回收
率(%)RSD (%) 血清1 2.8 5.0 105.6 2.8 10 87.0 3.4 15 97.2 2.3 血清2 2.6 5.0 93.0 2.3 10 97.1 1.1 15 99.7 5.7 血清3 3.1 5.0 102.4 1.7 10 95.7 4.6 15 93.2 2.5 
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