For such a simple technique, there's a lot that can go wrong with high-performance liquid chromatography (HPLC) (For a description of how next-generation HPLCs work, see How It Works). Pressure can go haywire, lines can get clogged, and of course, columns don't last forever. We asked Tom Jupille, president of LC Resources, a company that offers courses in HPLC diagnostics, troubleshooting, and preventative maintenance; Uwe Neue, a principal research chemist in the chemistry operations group at Waters Corp.; and Tom Wheat, principal scientist and manager of the life science application laboratory at Waters, for advice in maintaining and troubleshooting your HPLC. Here's what they said.
1. KNOW YOUR BUFFERS
"People have paid me exorbitant consulting fees, only to find that two labs are interpreting the same recipe differently," says Jupille.
HPLC relies on differential equilibria between your analyte, the stationary phase (column), and a mobile phase (solvent). "A common mobile ...
