PURIFICATION OF THYMINE DIMER FROM MONOMER SOLUTION IN PREPARATIVE LIQUID-CHROMATOGRAPHY

Cyclobutane pyrimidine dimers and monomers of thymine were separated o n a Cls reversed-phase high performance liquid chromatographic column. Using two mathematical models, the effect of the sample sizes on peak shapes in preparative liquid chromatography was investigated. One of these approaches is through a linear kinetic model, and the other is b ased on the nonlinear adsorption isotherm. With the injection of small samples good agreement between the calculated value by the linear kin etic model and experimental data were achieved. With increased sample size, this model is defective in its prediction of large concentration profiles of sample. However, the nonlinear model permits the accurate prediction of the location of the component band and the determinatio n of the appropriate time to start and stop collection of the enriched fraction at higher concentrations of monomer and the lower concentrat ions of dimer. Therefore, extremely small amounts of dimer can be extr acted from monomer solutions.