On-Chip Spyhole Nanoelectrospray Ionization Mass Spectrometry for Sensitive Biomarker Detection in Small Volumes

  • Xiaoqin Zhong
  • Liang Qiao
  • Géraldine Stauffer
  • Baohong Liu
  • Hubert H. Girault
Research Article


A polyimide microfluidic chip with a microhole emitter (Ø 10–12 μm) created on top of a microchannel by scanning laser ablation has been designed for nanoelectrospray ionization (spyhole-nanoESI) to couple microfluidics with mass spectrometry. The spyhole-nanoESI showed higher sensitivity compared to standard ESI and microESI from the end of the microchannel. The limits of detection (LOD) for peptide with the spyhole-nanoESI MS reached 50 pM, which was 600 times lower than that with standard ESI. The present microchip emitter allows the analysis of small volumes of samples. As an example, a small cell lung cancer biomarker, neuron-specific enolase (NSE), was detected by monitoring the transition of its unique peptide with the spyhole-nanoESI MS/MS. NSE at 0.2 nM could be well identified with a signal to noise ratio (S/N) of 50, and thereby its LOD was estimated to be 12 pM. The potential application of the spyhole-nanoESI MS/MS in cancer diagnosis was further demonstrated with the successful detection of 2 nM NSE from 1 μL of human serum. Before the detection, the serum sample spiked with NSE was first depleted with immune spin column, then desalted by centrifugal filter device, and finally digested by trypsin, without any other complicated preparation steps. The concentration matched the real condition of clinical samples. In addition, the microchips can be disposable to avoid any cross contamination. The present technique provides a highly efficient way to couple microfluidics with MS, which brings additional values to various microfluidics and MS-based analysis.

Graphical Abstract


Microfluidics Nanoelectrospray ionization Interface Biomarker 



Xiaoqin Zhong would like to acknowledge the China Scholarship Council for her PhD study scholarship. Liang Qiao would like to thank NSFC (81671849) and MOST (2016YFE0132400) for funding support.

Supplementary material

13361_2018_1937_MOESM1_ESM.docx (1024 kb)
ESM 1 (DOCX 1023 kb)


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Copyright information

© American Society for Mass Spectrometry 2018

Authors and Affiliations

  • Xiaoqin Zhong
    • 1
  • Liang Qiao
    • 2
  • Géraldine Stauffer
    • 1
  • Baohong Liu
    • 2
  • Hubert H. Girault
    • 1
  1. 1.Laboratoire d’Electrochimie Physique et AnalytiqueEcole Polytechnique Fédérale de LausanneSionSwitzerland
  2. 2.Chemistry Department, Fudan UniversityShanghaiChina

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