Computer simulation for the convenient optimization of isocratic reversed-phase liquid chromatographic separations by varying temperature and mobile phase strength

Software is described which allows the rapid development of separations by means of isocratic reversed-phase Liquid chromatography (RP-LC), based on t he optimization of column temperature (T) and mobile phase strength (%B). F or a given sample, four initial experiments are carried out at two differen t temperatures, using either isocratic or (better) gradient elution. if iso cratic experiments are chosen for computer simulation, it is necessary to s elect appropriate values of %B for these initial runs. Literature data for solute retention as a function of T are reviewed, as a basis for estimating suitable values of %B at the two values of T selected. The use of optimize d values of T and %B led to acceptable separations for three representative samples. The prediction of isocratic separation on the basis of initial gr adient experiments is more convenient than the use of initial isocratic exp eriments, but less reliable. When gradient experiments are used, one additi onal isocratic experiment can improve the accuracy of such predictions by a "reflection" procedure. The latter approach was confirmed for predictions of both isocratic and gradient separation from initial gradient experiments . (C) 2000 Elsevier Science B.V. All rights reserved.