Analytical and Bioanalytical Chemistry

, Volume 411, Issue 28, pp 7431–7440 | Cite as

Ratiometric detection of alkaline phosphatase based on aggregation-induced emission enhancement

  • Fei QuEmail author
  • Lingxin Meng
  • Yuqiu Zi
  • Jinmao You
Research Paper


Alkaline phosphatase (ALP) is an important enzyme that is associated with many human diseases, so the quantitative detection of ALP is vital from a clinical perspective. Nevertheless, most fluorescent assays for monitoring ALP depend on aggregation-induced quenching (ACQ), single-signal modulation, or a “signal off” mode, which suffer from poor sensitivity, a “false positive” problem, and low signal output. In this work, we utilized the electrostatically driven self-assembly of glutathione-capped gold nanoclusters (GSH-AuNCs, which show aggregation-induced emission, AIE) and amino-modified silicon nanoparticles (SiNPs) to create a hybrid probe (SiNPs@GSH-AuNCs). This nanohybrid probe showed emission from the SiNPs at around 470 nm as well as aggregation-induced emission enhancement (AIEE) of the GSH-AuNCs at 580 nm. The AIEE of the GSH-AuNCs was quenched in the presence of KMnO4, but the AIEE was recovered by adding ascorbic acid as an oxidation–reduction reaction occurred between KMnO4 and the ascorbic acid. The fluorescence of the SiNPs remained constant whether the AIEE was quenched or not, meaning that the fluorescence of the SiNPs could be used as an internal reference. In a typical enzymatic reaction, ascorbic acid 2-phosphate is hydrolyzed by ALP to produce ascorbic acid. Therefore, the hybrid probe was shown to allow the ratiometric detection of ALP, with a linear range of 0.5–10 U L−1 and a limit of detection (LOD) of 0.23 U L−1. Finally, the proposed analytical strategy was successfully applied to detect ALP in human serum samples and to determine the concentration of an ALP inhibitor.

Graphical Abstract


Alkaline phosphatase Aggregation-induced emission enhancement Oxidation-reduction reaction Gold nanoclusters Silicon nanoparticles 



This work was supported by the National Natural Science Foundation of China (Youth Fund Project) (21405093), the Natural Science Foundation of Shandong Province, China (ZR2019QB010), and the Scientific Research Foundation of Qufu Normal University (BSQD20130117).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical committee approval

The human serum sample experiments were performed in accordance with the guidelines from the Ethical Committee, Qufu Normal University. All serum samples were obtained from healthy volunteers with their informed consent. All studies were approved by the Ethical Committee of Qufu Normal University.

Supplementary material

216_2019_2098_MOESM1_ESM.pdf (2.5 mb)
ESM 1 (PDF 2538 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Fei Qu
    • 1
    • 2
    Email author
  • Lingxin Meng
    • 1
    • 2
  • Yuqiu Zi
    • 1
    • 2
  • Jinmao You
    • 1
    • 2
    • 3
  1. 1.The Key Laboratory of Life-Organic AnalysisQufu Normal UniversityQufuChina
  2. 2.The Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural MedicineQufu Normal UniversityQufuChina
  3. 3.Key Laboratory of Tibetan Medicine Research & Qinghai Provincial Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau BiologyChinese Academy of ScienceXiningChina

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