摘要

We have developed a novel technique for the absolute determination of the mobile phase volume stored inside of a variable volume pulse damper at different pressures. Using an Agilent HPLC pulse damper we found a linear volume increase of similar to 1 mu L/bar. We found that pump pressurization is a relatively slow process and takes similar to 1 min to reach 90% and takes similar to 2 min to reach 99% equilibration at flow rates below 1 mL/min. During pump pressurization, column flow rate will be less than the pre-set, since part of the mobile phase is retained inside of the pulse damper. During our experiments we observed a discrepancy between data obtained by UV techniques and direct absolute measurements. This difference can be explained by a fundamental difference between the gradient delay volume and dead (dwell) volume.

  • 出版日期2010-5