Nanoflow gradient generator for capillary high-performance liquid chromatography

Anal Chem. 2004 Mar 1;76(5):1524-8. doi: 10.1021/ac0350312.

Abstract

A novel method of generating a nanoflow gradient elution for a capillary high-performance liquid chromatography (HPLC) system has been developed. An important feature of this system is that any gradient (GR) profile generated by a conventional microflow GR pump can be asymptotically traced and converted as a corresponding nanoflow GR profile simply by using a 10-port switching valve with two injection loops installed. Consequently, it has been called an "asymptotic trace 10-port valve" (AT10PV) nanoflow GR generator. Performance of the AT10PV nanoflow GR generator was tested in the range of flow rates from 50 to 500 nL/min. The test demonstrated that the AT10PV nanoflow GR generator can asymptotically trace the original gradient profile with good reproducibility. A capillary HPLC system using the AT10PV nanoflow GR generator provides reasonably good repeatability of peak retention times on the chromatogram of the tryptic digest of a BSA sample, RSD of less than 0.3% at a flow rate of 200 nL/min. It also enables sequential running of a series of sample injections in the same manner as conventional analysis at microflow rates.

MeSH terms

  • Acetone
  • Acetonitriles
  • Chromatography, High Pressure Liquid / instrumentation*
  • Chromatography, High Pressure Liquid / methods*
  • Microfluidics*
  • Nanotechnology
  • Reproducibility of Results
  • Serum Albumin, Bovine / analysis
  • Serum Albumin, Bovine / metabolism
  • Solvents
  • Time Factors
  • Trypsin / metabolism

Substances

  • Acetonitriles
  • Solvents
  • Acetone
  • Serum Albumin, Bovine
  • Trypsin
  • acetonitrile