Suitability of stationary phase for LC analysis of biomolecules

Biologically active compounds such as carotenoids/isoprenoids, vitamins, steroids, saponins, sugars, long chain fatty acids, and amino acids play a very important role in coordinating functions in living organisms. Determination of those substances is indispensable in advanced biological sciences. Engineered stationary phase in LC for the analysis of biomolecules has become easier with the development of chromatographic science. In general, C₁₈ column is being used for routine analysis but specific columns are being used for specific molecule. Monolithic columns are found to have higher efficiency than normal column. Among recent introduction, triacontyl stationary phases, designed for the separation of carotenoid isomers, are widely used for the estimation of carotenoids. In comparison to conventional C₁₈ phases, C₃₀ phases exhibited superior shape selectivity for the separation of isomers of carotenoids. It is also found useful for better elution and analysis of tocopherols, vitamin K, sterols, and fatty acids. Vitamin K, E, and their isomers are also successfully resoluted and analyzed by using C₃₀ column. Amino bonded phase column is specifically used for better elution of sugars, whereas phenyl columns are suitable for the separation and analysis of curcuminoids and taxol. Like triacontyl stationary phase, pentafluorophenyl columns are also used for the separation and analysis of carotenoids. Similarly, HILIC column are best suited for sugar analysis. All the stationary phases are made possible to resolute and analyze the target biomolecules better, which are the future of liquid chromatography. The present article focuses on the differential interaction between stationary phase and target biomolecules. The applicability of these stationary phases are reported in different matrices.