Rapid quantitation of ultraviolet-induced thymine-containing dimers in human cell DNA by reversed-phase...

Rapid and efficient separation of all three types of cyclobutyl pyrimidine dimer (Pyr mean value of Pyr) species induced in cellular DNA by far-ultraviolet (UV) light (chiefly 254 nm) has been achieved by reversed-phase high-performance liquid chromatography using octadecylsilyl stationary phases. The order of elution is: (Ura mean value of Ura) less than (Ura mean value of Thy) less than (Thy mean value of Thy) less than Thy. The determination of Pyr mean value of Pyr species in DNA from UV-irradiated, [3H]thymidine-labelled human skin fibroblasts in tissue culture is demonstrated for far-UV fluences as low as 10 J/m2. The ability to measure specifically individual dimer types allowed demonstration of comparable kinetics of repair for two labelled dimer species (Ura mean value of Thy and Thy mean value of Thy).