Combined use of temperature and solvent strength in reversed-phase gradient elution. I. Predicting separation as a function of temperature and gradient conditions.
It has been shown previously that computer simulation based on two initial experiments can predict separation in reversed-phase gradient elution as a function of gradient conditions (gradient steepness, gradient range and gradient shape) and column conditions (column length, flow-rate and particle size). The present study extends this capability for changes in temperature. Four initial experiments (two different gradient times, two different temperatures) provide input data that allow predictions of separation as a function of temperature as well as gradient and column conditions. A semi-empirical relationship, tR = a + bT, is able to relate gradient retention time tR to column temperature T (other conditions constant). The accuracy of this approach has been evaluated for 102 solutes and a variety of experimental conditions, including the use of five different HPLC instruments (four different models).
Zhu
P L
PL
LC Resources Inc., Walnut Creek, CA 94596, USA.
Snyder
L R
LR
Dolan
J W
JW
Djordjevic
N M
NM
Hill
D W
DW
Sander
L C
LC
Waeghe
T J
TJ
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D012997:Solvents; D013696:Temperature
D012997:Solvents