ANYL 31 |
| Proteomic samples, especially peptides from protein enzymatic digests, are most commonly separated by reversed-phase gradient elution chromatography. Due to the large number of components in a single sample, optimizing the peak capacity of such separation is of great practical importance. In this work, we will use a Monte Carlo approach to study the effect of different operational variables (e.g. gradient time and temperature) on the peak capacity of peptide separations. Once the best initial and final organic modifier fraction to maximize the retention difference between the first and last eluting peptides is found, flow rate, temperature and gradient time are simultaneously varied to optimize peak capacity. We found that peak capacity is maximized at the optimum flow rate on a given column and at longer gradient times while temperature has a much smaller impact. In addition, an optimum range gradient steepness exists where peak capacity is maximized. Finally, these conclusions are verified by experiments that are in very good agreement with the theory. |
|
General Papers
8:30 AM-11:10 AM, Sunday, 28 August 2005 Washington DC Convention Center -- 153, Oral
Sci-Mix
Division of Analytical Chemistry |