Nanofractionation spotter technology for rapid contactless and high-resolution deposition of LC eluent for further off-line analysis

Anal Chem. 2011 Jan 1;83(1):125-32. doi: 10.1021/ac102001g. Epub 2010 Nov 30.

Abstract

The development of a contactless postcolumn spotter technology capable of rapidly and accurately depositing LC eluent onto another platform (e.g., 1536-well microtiter plates) is described. Many detection methodologies are suitable for online analysis, such as mass spectrometry, UV-vis, and fluorescence. In some cases, when online analysis is less suitable, off-line postcolumn analysis is the methodology of choice and usually relies on LC-based fractionation prior to detection (e.g., MALDI-MS, Raman spectrsocopy, biochemical assays). As fractionation generally involves loss in resolution, the technology described here allows high-resolution contactless fractionation by tailoring the fractionation frequency to the chromatographic peaks and mixing in of postcolumn reagents. Droplet ejection at frequencies of at least 6 Hz could be performed in the nanoliter to low microliter range with repeatabilities of ∼6%. Furthermore, multiple droplets can be ejected at the same position thereby allowing adjustment of fractionation volume and speed. The technology was evaluated, optimized, and validated prior to two proof-of-principle demonstrations comprising off-line chemical detection of injected fluorescein and off-line postcolumn biochemical detection of acetylcholine-binding protein ligands, both based on 1536-well plate reader analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carrier Proteins / isolation & purification
  • Carrier Proteins / metabolism
  • Cell Line
  • Chemical Fractionation / instrumentation
  • Chemical Fractionation / methods*
  • Chromatography, Liquid / methods*
  • Fluorescein / chemistry
  • Indicators and Reagents / chemistry
  • Mass Spectrometry
  • Nanotechnology / instrumentation
  • Nanotechnology / methods*
  • Reproducibility of Results
  • Time Factors

Substances

  • AChBP protein, Lymnaea
  • Carrier Proteins
  • Indicators and Reagents
  • Fluorescein