Tributyl Phosphate as Stationary Phase in Reversed Phase Liquid Chromatographic Separations of Hydrophilic Carboxylic Acids, Amino Acids and Dipeptides

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Abstract
Tributyl phosphate (TBP) was used as stationary liquid phase in reversed phase liquid column chromatography with aqueous buffers or aqueous buffers + methanol as the mobile phase.Hydrophobic microparticulate (5 .mu.m) alkyl-derivatized silica was used as support. The stability of the columns is very high as is their separating efficiency. The strong H accepting properties of TBP makes retention possible of highly polar solutes containing H donating groups. Retention of carboxylic acids is obtained by the distribution of the uncharged form into TBP and it is regulated by the pH of the mobile phase. No influence from the support on the retention of carboxylic acids has been seen. Benzoic, phenylacetic, mandelic, IAA and glucuronic acid derivatives, amino acids and dipeptides were chromatographed and the influence of structural effects on the separation factors is discussed.