谷氨酸和组氨酸功能化石墨烯量子点的合成及用于肿瘤细胞中过氧化氢比色测定与荧光成像

引用本文: 江燕红, 李娜娜, 李在均. 谷氨酸和组氨酸功能化石墨烯量子点的合成及用于肿瘤细胞中过氧化氢比色测定与荧光成像[J]. 分析化学, 2021, 49(5): 809-819. doi: 10.19756/j.issn.0253-3820.201560 shu

Citation: JIANG Yan-Hong, LI Na-Na, LI Zai-Jun. Synthesis of Glutamic Acid and Histidine Functionalized Graphene Quantum Dots and Their Application as a Peroxidase for Colorimetric Determination of H₂O₂ Inside and Outside Tumor Cells and Fluorescence Imaging[J]. Chinese Journal of Analytical Chemistry, 2021, 49(5): 809-819. doi: 10.19756/j.issn.0253-3820.201560 shu

谷氨酸和组氨酸功能化石墨烯量子点的合成及用于肿瘤细胞中过氧化氢比色测定与荧光成像

江南大学化学与材料工程学院, 无锡 214122

通讯作者: 李在均,E-mail:zaijunli@jiangnan.edu.cn

基金项目:
国家重点研发计划项目（No.2018YFC1603001）和国家自然科学基金项目（No.21576115）资助。

摘要: 通过热解柠檬酸、谷氨酸和组氨酸的混合物得到谷氨酸和组氨酸功能化石墨烯量子点（Glu-GQD-His）。此量子点由2~3层石墨烯片组成，平均片径为（3.85±0.15）nm，含有丰富的功能基团，在可见光激发下发出强的蓝色荧光。Glu-GQD-His的发光效率高于单独柠檬酸合成的石墨烯量子点（GQD），用于细胞荧光成像时，观察到强蓝色荧光，说明谷氨酸和组氨酸的引入改善了GQD的光学性质。研究表明，Glu-GQD-His对H₂O₂氧化3，3'，5，5'-四甲基联苯胺生成蓝色氧化产物的反应具有明显的催化作用，催化反应的米氏常数（K_m）为1.110 mmol/L，低于辣根过氧化物酶（3.702 mmol/L）和GQD（1.443 mmol/L），说明Glu-GQD-His具有更高的催化活性。基于此催化活性，建立了测定H₂O₂的比色分析方法。H₂O₂浓度在0.001~1.0 mmol/L范围内，反应体系的吸光度随H₂O₂浓度的增加而线性增加，检出限为0.4 μmol/L，灵敏度和选择性优于现有H₂O₂比色方法。本方法成功用于HepG2细胞内外H₂O₂的测定。

关键词:

English

Synthesis of Glutamic Acid and Histidine Functionalized Graphene Quantum Dots and Their Application as a Peroxidase for Colorimetric Determination of H₂O₂ Inside and Outside Tumor Cells and Fluorescence Imaging

School of Chemical and Material Engineering, Jiangnan University, Wuxi 214122, China

Corresponding author: LI Zai-Jun, zaijunli@jiangnan.edu.cn

Received Date: 07 September 2020
Revised Date: 03 February 2021
Fund Project:
Supported by the National Key Research and Development Program of China (No.2018YFC1603001) and the National Natural Science Foundation of China (No.21576115).

Abstract: Glutamic acid and histidine functionalized graphene quantum dots (Glu-GQD-His) were synthesized by pyrolysis of the mixture of citric acid, glutamic acid and histidine. The resulting Glu-GQD-His was composed of 2-3 layers of graphene sheets with an average size of (3.85±0.15) nm and rich functional groups. Glu-GQD-His could offer a strong blue fluorescence emission under visible light excitation, which was higher than that of graphene quantum dot (GQD) made via pyrolysis of citric acid. The results demonstrated that the introduction of glutamic acid and histidine could improve the optical properties. As an optical probe, Glu-GQD-His was of a low toxicity and could be used for cell fluorescence imaging. As an artificial peroxidase, Glu-GQD-His could catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine with H₂O₂ to produce corresponding blue oxidation product. The Michaelis constant (K_m) reached 1.110 mmol/L, which was significantly lower than that of horseradish peroxidase (3.702 mmol/L) and GQD (1.443 mmol/L). This verified that Glu-GQD-His had a better catalytic activity. Based on the above catalytic reaction, the colorimetric method for determination of H₂O₂ was established. In the concentration range of 0.001-1.0 mmol/L, the absorbance of blue oxidation products would increase linearly with increasing H₂O₂ concentration. The detection limit of the method was 0.4 μmol/L. The as-proposed method provided better sensitivity and selectivity compared with the present analytical methods, and was successfully applied to detection of H₂O₂ inside and outside tumor cells.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

谷氨酸和组氨酸功能化石墨烯量子点的合成及用于肿瘤细胞中过氧化氢比色测定与荧光成像

通讯作者: 李在均,E-mail:zaijunli@jiangnan.edu.cn

English

Synthesis of Glutamic Acid and Histidine Functionalized Graphene Quantum Dots and Their Application as a Peroxidase for Colorimetric Determination of H₂O₂ Inside and Outside Tumor Cells and Fluorescence Imaging

Corresponding author: LI Zai-Jun, zaijunli@jiangnan.edu.cn

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CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

谷氨酸和组氨酸功能化石墨烯量子点的合成及用于肿瘤细胞中过氧化氢比色测定与荧光成像

通讯作者: 李在均,E-mail:zaijunli@jiangnan.edu.cn

English

Synthesis of Glutamic Acid and Histidine Functionalized Graphene Quantum Dots and Their Application as a Peroxidase for Colorimetric Determination of H2O2 Inside and Outside Tumor Cells and Fluorescence Imaging

Corresponding author: LI Zai-Jun, zaijunli@jiangnan.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

Synthesis of Glutamic Acid and Histidine Functionalized Graphene Quantum Dots and Their Application as a Peroxidase for Colorimetric Determination of H₂O₂ Inside and Outside Tumor Cells and Fluorescence Imaging

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs