Quantitative Liquid Column and Thin Layer Chromatography of Lipids and Other Water Insoluble Substances, Elution Selectivity Principles, and a Graphic Method for Pattern Analysis of Chromatographic Data
The classification of chromatographic solvents according to the nature of their hydrogen bonding, acidic, and basic groups is considered in the light of solvent-solute interactions, solubility, and solvent selectivity in liquid column chromatography (LCC) and thin layer chromatography (TLC). The effects on chromatographic migration of the different types of hydrogen bonding groups (proton donor only, proton acceptor only, both proton donor and acceptor), acids, and bases are illustrated and interpreted. Solvent systems are presented for the separation of lipids and other water insoluble organic compounds over a broad range of functional groups. The special advantages of TLC and LCC are pointed out as are the ways high speed can be obtained with both methods without loss of resolution, accuracy, or precision of analysis. Also considered are the most accurate and precise general approaches to quantitative TLC and LCC, as well as automation of LCC and semiautomation of TLC using autoanalyzers with various reagents which produce derivatives that absorb light or fluoresce. Pattern analysis of a metabolically or functionally related group of biological compounds is defined as the determination of the relationship of each component to all others and to the total molar amount of all components. The requirements for accurate and precise pattern analysis are discussed, and the uses of a new graphic method for pattern analysis are demonstrated with data for organ and blood plasma lipids. The results demonstrate the need for highly accurate and precise data obtained by carefully proven methods of analysis.