Issue 22, 2014

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Abstract

Analytical technologies that enable investigations at the single cell level facilitate a range of studies; here a lab-fabricated capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) platform was used to analyze anionic metabolites from individual Aplysia californica neurons. The system employs a customized coaxial sheath-flow nanospray interface connected to a separation capillary, with the sheath liquid and separation buffer optimized to ensure a stable spray. The method provided good repeatability of separation and reliable detection sensitivity for 16 mono-, di- and triphosphate nucleosides. For a range of anionic analytes, including cyclic adenosine monophosphate (cAMP), adenosine diphosphate (ADP) and adenosine triphosphate (ATP), the detection limits were in the low nanomolar range (<22 nM). A large Aplysia R2 neuron was used to demonstrate the ability of CE-ESI-MS to quantitatively characterize anionic metabolites within individual cells, with 15 nucleotides and derivatives detected. Following the method validation process, we probed smaller, 60 μm diameter Aplysia sensory neurons where sample stacking was used as a simple on-line analyte preconcentration approach. The calculated energy balance ([ATP] + 0.5 × [ADP])/([AMP] + [ADP] + [ATP]) of these cells was comparable with the value obtained from bulk samples.

Graphical abstract: Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

Supplementary files

Article information

Article type
Paper
Submitted
23 Jun 2014
Accepted
28 Aug 2014
First published
29 Aug 2014

Analyst, 2014,139, 5835-5842

Analysis of endogenous nucleotides by single cell capillary electrophoresis-mass spectrometry

J. Liu, J. T. Aerts, S. S. Rubakhin, X. Zhang and J. V. Sweedler, Analyst, 2014, 139, 5835 DOI: 10.1039/C4AN01133C

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