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Fluorescent sensing of free bilirubin at nanomolar level using a Langmuir–Blodgett film of glucuronic acid–functionalized gold nanoclusters

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Abstract

Serum bilirubin is an important indicator to assess liver function and diagnose various types of liver diseases. The level of serum bilirubin is also negatively correlated with the risk of cardiovascular disease and cancer. We had fabricated a fluorescent film sensor aiming at free bilirubin detection at the nanomolar level. Gold nanoclusters capped by human serum albumin (HSA–AuNCs) were utilized as a fluorescent platform for bilirubin biorecognition. HSA–AuNCs were functionalized with glucuronic acid to increase the binding sites for bilirubin. An ultrathin film of glucuronic acid–functionalized gold nanoclusters was obtained by the Langmuir-Blodgett (LB) technique. When exposed to bilirubin, the interaction between free bilirubin and the functionalized AuNCs resulted in fluorescent quenching of the film. Good linearity could be achieved for the quenching efficiency versus the logarithm of free bilirubin concentration over a concentration range of 1.00 nM~5.00 μM. The limit of detection (LOD) was calculated to be (2.70 ± 0.14) × 10−1 nM (S/N = 3). The film sensor presents a good anti-interference capability towards common substances coexisting with bilirubin in serum. Satisfactory results achieved in the tests of real serum samples indicate that the LB film sensor can be used for bilirubin determination in nanomolar concentration.

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Acknowledgements

This research was financially supported by the National Natural Science Foundation of China (No. 61761013 and 61864001), the Natural Science Foundation of Guangxi Province (No. 2017GXNSFAA198116), and the Guangxi Key Laboratory of Automatic Detection Technology and Instruments Foundation Project (No. YQ17113).

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  1. School of Life and Environmental Sciences, Guilin University of Electronic Technology, Guilin, 541004, Guangxi, China

    Wenxiang Xiao, Jing Liu, Yinan Xiong, Yaoxin Li & Hua Li

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  1. Wenxiang Xiao
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  2. Jing Liu
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  4. Yaoxin Li
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  5. Hua Li
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Correspondence to Wenxiang Xiao or Hua Li.

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The human serum samples used in this study were approved by the Guangxi Key Laboratory of Metabolic Diseases Research Ethics Committee in Guilin, China.

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Xiao, W., Liu, J., Xiong, Y. et al. Fluorescent sensing of free bilirubin at nanomolar level using a Langmuir–Blodgett film of glucuronic acid–functionalized gold nanoclusters. Anal Bioanal Chem 413, 7009–7019 (2021). https://doi.org/10.1007/s00216-021-03660-6

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  • DOI: https://doi.org/10.1007/s00216-021-03660-6

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