Abstract
Biologically important substances have been widely analyzed by aqueous phase liquid chromatography. The systems are called reversed-phase or ion-exchange liquid chromatography. The optimization of the liquid chromatography of biomedically important compounds is very difficult. There is no simple solution or mathematical equations for separating complicated mixtures in liquid chromatography. When polar compounds are chromatographed on a hydrophobic packing with an aqueous eluent, the chromatography is called reversed-phase liquid chromatography. The elution order can be related to the hydrophobicity of the compounds. Further, the hydrophobicity is related to the partition coefficients of the solutes. The prediction of the retention times of partly ionized aromatic acids is possible from their hydrophobic fragmental constants (log P) and their dissociation constants (pK a ). The partition coefficients of several compounds have been directly measured in liquid chromatography and related to the observed capacity factors. The liquid chromatographic system uses octadecyl-bonded silica gels as the packing and pH-controlled acetonitrile-water mixtures as the eluent. According to Rekker's method, the partition coefficient (log P) can be calculated as a sum of hydrophobic fragmental constants.
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