Multi-²D LC × LC as a Novel and Powerful Implement for the Maximum Separation of Complex Samples

Achieving complete information about the chemical composition of complex samples requires the use of multianalytical platforms able to maximize the acquisition of high-quality data for unequivocal identification. However, this process requires long analysis times and several instruments. Food analysis is one of the analytical fields where the analysis of very complex samples has a huge impact. One of these complex samples is vermouth, a fortified wine based on the maceration of a large number of herbs, fruits, barks, seeds, and leaves. The application of conventional or even advanced analytical techniques like comprehensive two-dimensional (2D) liquid chromatography (LC × LC) does not provide enough separation power to resolve the complete profile of this sample. In this work, a novel 2DLC strategy called multi-²D LC × LC is developed. This new setup consists of the use of two different columns with different separation properties in the second dimension (²D) that can be selected during the LC × LC analysis accordingly to the chemical nature of the compounds eluted from the first dimension (¹D). The vermouth sample was analyzed using a ¹D-PFP and a combination of HILIC (from 0 to 30 min) and C18 (from 30 to the end) columns in the ²D. This setup increased both the peak capacity and the orthogonality of the analysis in comparison to the use of only one of the columns in the ²D. Multi-²D LC × LC is presented as an integrated 2DLC tool that maximizes the separation capacity for very complex samples.