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Nitrogen-doped carbon quantum dots fabricated from cellulolytic enzyme lignin and its application to the determination of cytochrome c and trypsin

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Abstract

A sensitive and effective strategy for the detection of cytochrome c (Cyt c) and trypsin was developed using biomass nitrogen-doped carbon quantum dots (N-CQDs) as the fluorescence probe. N-CQDs were synthesized through a one-pot hydrothermal method by utilizing cellulolytic enzyme lignin as the carbon source and ammonia as the solvent and nitrogen source. The obtained N-CQDs had good water solubility and stable optical properties. The introduction of nitrogen increased fluorescence quantum yield (QY) to 8.23%, which was almost four times as high as that before nitrogen doping. The N-CQDs were fabricated as a label-free biosensor to detect Cyt c and trypsin. The fluorescence of N-CQDs was quenched with positively charged Cyt c due to electrostatic induction aggregation and static quenching. However, Cyt c tended to be hydrolyzed into small peptides in the presence of trypsin, which caused fluorescence recovery of the N-CQDs/Cyt c complex. A wide linear response range was achieved for Cyt c within 1–50 μM and the developed N-CQDs/Cyt c complex displayed a linear response for trypsin within 0.09–5.4 U/mL. The detection limits were 0.29 μM for Cyt c and 0.013 U/mL for trypsin, respectively. Furthermore, this assay had been applied to Cyt c and trypsin detection in serum samples with the recoveries in the range of 94.6–98.5% and 95.5–102.0%, respectively. The established method was sensitive, selective, easy to operate, and low cost, which proved its potential application in clinical diagnosis.

Graphical abstract

The synthesis and fluorescence mechanism of N-CQDs and the strategy for Cyt c and trypsin detection

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Funding

This project was funded by Natural Science Foundation of Heilongjiang Province (B2008001), Heilongjiang Postdoctoral Fund (LBH-Z16009), 111 Project (B20088), Heilongjiang Touyan Innovation Team Program (Tree Genetics and Breeding Innovation Team), and China Postdoctoral Science Foundation (2016M591501, 2017T100218).

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Authors and Affiliations

  1. College of Chemistry, Chemical Engineering and Resource Utilization, Key Laboratory of Forest Plant Ecology, Northeast Forestry University, 26 Hexing Road, Harbin, 150040, Heilongjiang, China

    Chenhui Yin, Ligang Chen & Na Niu

  2. Key Laboratory of Superlight Materials and Surface Technology, Ministry of Education, College of Materials Science and Chemical Engineering, Harbin Engineering University, Harbin, 150001, Heilongjiang, China

    Na Niu

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  1. Chenhui Yin
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Correspondence to Ligang Chen or Na Niu.

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All applicable international, national, and/or institutional guidelines for the collection and use of human blood and serum samples were followed. All biological experiments were approved by the Ethics Committee of Harbin Medical University Cancer Hospital.

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Yin, C., Chen, L. & Niu, N. Nitrogen-doped carbon quantum dots fabricated from cellulolytic enzyme lignin and its application to the determination of cytochrome c and trypsin. Anal Bioanal Chem 413, 5239–5249 (2021). https://doi.org/10.1007/s00216-021-03496-0

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