6.2-20
Section 6
ApplicAtions
Pinnacle Operators Manual
Pickering Laboratories Inc.
Make only one small change at a time.
Make a change only after you have collected at least two chromatograms showing the same separation. This
usually means three injections, as the first injection of a series rarely is representative of the rest of the series.
Optimize the separations in the early part of the gradient before optimizing the late part.
Every model of HPLC forms gradients differently. The programs suggested in this manual or in the information
sheets are typical of the more popular HPLC pumps. Consult Pickering Laboratories if you need advice.
If you need only the early part of the chromatogram, you can save time by truncating the gradient. Go to the
final concentration of regenerant and hold it until the most basic component (arginine) elutes, then
re-equilibrate with the initial buffer.
The separation is temperature sensitive. Adjusting the temperature may improve it. For example, the resolution
of threonine and serine improves when the column temperature is cooler, however the resolution
of tyrosine and phenylalanine is best when the column temperature is warmer.
Surfactants, dyes, ninhydrin, and lipids usually cannot be removed. Prevention is the only cure.
To Next Page
Loading...