Text Box: RECENT ADVANCES IN COMPREHENSIVE TWO-DIMENSIONAL LIQUID CHROMATOGRAPHY FOR THE ANALYSIS OF NATURAL PRODUCTS FRANCESCO CACCIOLA1, PAOLA DONATO2, LUIGI MONDELLO1,2, AND PAOLA DUGO1,2 1DIPARTIMENTO FARMACO-CHIMICO, FACOLTĄ DI FARMACIA, UNIVERSITĄ DI MESSINA, VIALE ANNUNZIATA, 98168 MESSINA, ITALY AND 2 UNIVERSITĄ CAMPUS BIO-MEDICO, VIA ĮLVARO DEL PORTILLO 21, 00128 - ROMA, ITALY Introduction The analysis of real matrices, namely biological, food and environmental samples, poses a high demand for HPLC methods enabling determination and identification of a large number of compounds occurring in the samples. However, a single column does not often have sufficient separation power for the baseline separation of all components in complex samples, and the extent of peak overlap is enhanced as the number of compounds increases. The development of comprehensive two-dimensional high performance liquid chromatography (2D-LC) has been relatively slow until recently, even though the advantages of the technique were already described in the late 1970s and early 1980s (Karger et al., 1973; Erni & Frei, 1978; Giddings, 1984). In the 1990s the application of LCxLC was limited to proteomic applications, but in the last decade the utility of this technique in the separation of a variety of other complex mixtures has been demonstrated. The general motivation which lies behind this research area is linked to its potential for substantially more resolving (separating) power in comparison with the conventional one Dimensional (1D) counterparts. The most general set-up of a comprehensive LC system consists of two pumps, two columns, injector, interface and detector. The interface is in general a high pressure switching valve, and this device is often referred to as a modulator or sampling device. The suitability of any LC separation system for resolving complex samples can be expressed by theoretical peak capacity, nc, which determines the maximum number of peaks that can be separated side by side, into the separation space at a desired degree of resolution, e.g., RS = 1 (nc = t/w).

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