Combined effect of solvent content, temperature and pH on the chromatographic behaviour of ionisable compounds II: benefits of the simultaneous optimisation.
A previously reported eight-parameter mechanistic model [Part I of this work, J. Chromatogr. A 1163 (2007) 49] was applied to optimise the separation of 11 ionisable compounds (nine diuretics and two beta-blockers), considering solvent content, temperature and pH as experimental factors. The data from 21 experiments, arranged in a central composite design, were used to model the retention. Local models were used to predict efficiency and peak asymmetry. The optimisation strategy, based on the use of peak purity as chromatographic objective function and derived concepts, was able to find the most suitable experimental conditions yielding full resolution in reasonable analysis times. It also allowed a detailed inspection of the separation capability of the studied factors, and of the consequences of the shifts in the protonation constants originated by changes in solvent content and temperature. The size of the resolution structures suggested that the ranked importance of the factors was pH, organic solvent and temperature, giving rise to relatively narrow domains of full resolution. The three factors were found, however, worthwhile in the optimisation of selectivity. Predicted optimal conditions corresponding to two different optimal resolution regions were verified experimentally. In spite of the difficulties associated to the use of pH as optimisation factor, satisfactory agreement was found in both cases.
Pous-Torres
S
S
Departament de Química Analítica, Universitat de València, c/Dr. Moliner 50, 46100 Burjassot, Spain.
Torres-Lapasió
J R
JR
Baeza-Baeza
J J
JJ
García-Alvarez-Coque
M C
MC
D006863:Hydrogen-Ion Concentration; D012997:Solvents / Q000737:chemistry*; D013696:Temperature
D012997:Solvents