Theoretical study of incomplete sampling of the first dimension in comprehensive two-dimensional chromatography

Abstract

The process of regularly transferring material from the primary column to the secondary column is critical in producing comprehensive two-dimensional separations. A series of calculations have been performed to determine how sampling period, duty cycle, and sampling phase affect (1) the fraction of material transferred from the primary column to the secondary column, (2) the accuracy of primary retention time determination, and (3) the effective peak width along the primary retention axis. The results demonstrate that comprehensive two-dimensional separations can be produced without a substantial loss in quantitative precision and with only a moderate loss in primary column resolution if the sampling period is less than 1.5 times the primary peak standard deviation. The quantitative precision of total peak areas (for duty cycles less than 1.0) and primary retention time determination are rapidly reduced as the sampling period is increased above 1.5 times the primary peak standard deviation.