Direct Comparison of 75 micron Nanocolumns to Analytical Columns Using a Waters 2487 UV Detector Equipped with a Nanoliter Flow Cell under Isocratic Conditions

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Jennifer H Granger, Chris Stumpf, Rob Plumb, Liz Robertson, Dennis DellaRovere, Jeffrey W Finch, Steve Cohen
HPLC 2002; Montreal; 5-6 June 2002
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Nano-scale chromatography has gained much recent interest since it is readily coupled to mass spectrometry and allows routine analysis of nanomolar samples. However, use of more conventional HPLC detectors, such as UV detection, that are capable of handling nano-scale flow streams has been challenging. These nano-columns are almost exclusively operated under gradient elution conditions, e.g., peptide analysis, where true column performance is not readily measured. We have recently explored isocratic separations of small molecules on the nano-scale using 75 micron ID capillary columns packed with Waters Symmetry C18 stationary phases with UV detection via a Waters 2487 UV detector with either a 1 nL or 25 nL flow cell. A direct performance comparison can subsequently be made to an analytical scale Symmetry C18 (2.1 x 100 mm) column and a conventional UV flow cell. In this paper we will illustrate that 75 micron x 100 mm columns can be routinely packed with efficiencies greater than 110,000 plates per meter. We will also discuss how the chromatographic performance is affected by the type of unions employed and how band dispersion can be controlled. Finally we will show that the new nanoliter flow cell UV can be used prior to MS detection with no effect on the chromatographic peak shape.

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