Microchimica Acta

, 186:43 | Cite as

A split aptamer-labeled ratiometric fluorescent biosensor for specific detection of adenosine in human urine

  • Junhui You
  • Zhengyi You
  • Xin Xu
  • Jiangrong Ji
  • Tian Lu
  • Yuhong Xia
  • Lipin Wang
  • Liying Zhang
  • Shuhu Du
Original Paper


A dual-emission ratiometric fluorometric aptasensor is presented for highly specific detection of adenosine. An adenosine binding aptamer (ABA) was split into two halves (termed as ABA1 and ABA2). ABA1 was covalently bound to blue-emitting carbon dots (with excitation/emission maxima at 365/440 nm) as responsive fluorophore (referred to as ABA1-CDs). ABA2 was linked to red-emitting silica-coated CdTe quantum dots (with excitation/emission maxima at 365/613 nm) acting as internal reference and referred to as ABA2-QDs@SiO2. Upon addition of graphene oxide, the fluorescence of ABA1-CDs is quenched. After subsequent addition of ABA2-QDs@SiO2 and different amounts of adenosine, the blue fluorescence is recovered and causes a color change from red to royal blue. The method represents a ratiometric turn-on assay for visual, colorimetric and fluorometric determination of adenosine. The limit of detection is as low as 2.4 nM in case of ratiometric fluorometry. The method was successfully applied to the determination of adenosine in (spiked) human urine. Recoveries range from 98.8% to 102%.

Graphical abstract

Adenosine binding aptamer1-carbon dots (ABA1-CDs) can absorb on graphene oxide (GO) via π stacking. This causes fluorescence to be quenched by fluorescence resonance energy transfer (FRET). After addition of ABA2-silica-coated quantum dots (ABA2-QDs@SiO2) and adenosine, binding of adenosine to two pieces of aptamers forms a complex (ABA1-CD/adenosine/ABA2-QD@SiO2) which dissociates from GO. As a result, fluorescence is recovered.


Nanoprobe Fluorescence resonance energy transfer Carbon dots CdTe quantum dots Graphene oxide Visual detection Adenosine 



This work is supported by the National Natural Science Foundation of China (Nos. 21775073 and 61605084), Natural Science Foundationn of the Jiangsu Higher Education Institutions of China (No. 16KJB150029) and Science and Technology Funds of Nanjing Medical University (No. 2015NJMUZD023).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_3162_MOESM1_ESM.pdf (644 kb)
ESM 1 (PDF 643 kb)


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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Junhui You
    • 1
  • Zhengyi You
    • 1
  • Xin Xu
    • 1
  • Jiangrong Ji
    • 1
  • Tian Lu
    • 1
  • Yuhong Xia
    • 1
  • Lipin Wang
    • 1
  • Liying Zhang
    • 1
  • Shuhu Du
    • 1
  1. 1.School of PharmacyNanjing Medical UniversityNanjingChina

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