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Nanoflow Gradient Generator for CapillaryHigh-Performance Liquid Chromatography

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A novel method of generating a nanoflow gradient elutionfor a capillary high-performance liquid chromatography(HPLC) system has been developed. An important featureof this system is that any gradient (GR) profile generatedby a conventional microflow GR pump can be asymptotically traced and converted as a corresponding nanoflowGR profile simply by using a 10-port switching valve withtwo injection loops installed. Consequently, it has beencalled an “asymptotic trace 10-port valve” (AT10PV)nanoflow GR generator. Performance of the AT10PVnanoflow GR generator was tested in the range of flowrates from 50 to 500 nL/min. The test demonstrated thatthe AT10PV nanoflow GR generator can asymptoticallytrace the original gradient profile with good reproducibility. A capillary HPLC system using the AT10PV nanoflow GR generator provides reasonably good repeatabilityof peak retention times on the chromatogram of the trypticdigest of a BSA sample, RSD of less than 0.3% at a flowrate of 200 nL/min. It also enables sequential running ofa series of sample injections in the same manner asconventional analysis at microflow rates.

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