Sensitive Detection of Uric Acid Based on BSA Gold Nanoclusters by Fluorescence Energy Resonance Transfer
CONG Jian-han1, LUO Yun-jing1*, QI Xiao-hua2, ZOU Ming-qiang2, KONG Chen-chen1
1. Beijing Key Laboratory of Environmental and Viral Oncology, Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, China
2. China Academy of Inspection and Quarantine, Beijing 100123, China
Abstract:Based on gold nanoclusters (AuNCs) with good optical stability, biocompatibility and simple and non-toxic preparation method, this paper developed a highly selective, highly sensitive and visualized uric acid (UA) sensor. We synthesized BSA-AuNCs using bovine serum albumin (BSA) as a template. Under the catalysis of urate oxidase, UA produces stoichiometric hydrogen peroxide (H2O2), which causes the fluorescence of AuNCs to be quenched. In addition, we found that BSA-AuNCs mimic the peroxidase activity in this system, catalysing the substrate’s oxidation 3,3’,5,5’-tetramethylbenzidine (TMB) to ox-TMB by H2O2. At this time, the emission spectrum of BSA-AuNCs overlaps the absorption spectrum of ox-TMB, and a kind of fluorescence resonance energy transfer (FRET) occurs. BSA-AuNCs acts as a donor to transfer the excitation energy to the acceptor ox-TMB, which makes ox-TMB produce fluorescence. At the same time, the fluorescence intensity of BSA-AuNCs is significantly lower than when it exists alone, which significantly improves the sensitivity of UA detection. Under optimum conditions, it is found that the UA concentration has an excellent linear relationship between the quenching degree of 2~100 μmol·L-1, the linear equation is (F0-F)/F0=0.005 85cUA+0.103 64, the linear correlation coefficient is 0.995 4, and the detection limit is 0.26 μmol·L-1, which is far below the minimum limit of normal human UA levels (90 μmol·L-1). Meanwhile, the recovery of UA in blood samples was studied, and the recovery rate was 97.3% to 104.7%, indicating that this method is effective in clinical blood samples. The detection has tremendous application potential, and provides an excellent theoretical basis and methodological guidance for further clinical analysis.
Key words:Gold nanoclusters; Uric acid; Hydrogen peroxide; Fluorescence resonance energy transfer
丛剑涵,罗云敬,齐小花,邹明强,孔陈晨. 牛血清蛋白纳米金团簇基于荧光共振能量转移法灵敏检测尿酸[J]. 光谱学与光谱分析, 2022, 42(02): 483-489.
CONG Jian-han, LUO Yun-jing, QI Xiao-hua, ZOU Ming-qiang, KONG Chen-chen. Sensitive Detection of Uric Acid Based on BSA Gold Nanoclusters by Fluorescence Energy Resonance Transfer. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2022, 42(02): 483-489.
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