Normal-phase liquid chromatography compositional analysis

Normal-phase (NP) and reversed-phase (RP) liquid chromatography are simple divisions of the LC techniques based on the relative polarities of the mobile and stationary phases (Figure 4.10). Both NPLC and RPLC analysis make use of either the isocratic or gradient elution modes of separation (i.e. constant or variable composition of the mobile phase, respectively). Selection from these four available separation techniques depends on many variables but basically on the number and chemical structure of the compounds to be separated and on the scope of the analysis. 

Reversed-phase high-performance liquid chromatography (RP-HPLC) is more popular for analysis of carotenoids than is normal-phase HPLC because (1) retention is very little affected by small variations in the mobile-phase composition, and (2) the risk of artifact formation on passage through the column is minimal as solute-support interactions on non-polar-bonded phases only involve weak forces. A variety of stationary phases of various polarities are available, such as C18, C8, C4, C2, Cl, phenyl, and cyano derivatives the C18 phase is the most popular. 

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