COMPARISON OF EXTRACTION PROCEDURES FOR HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF CELLULAR DEOXYNUCLEOTIDES

For the determination of cellular deoxynucleoside triphosphates (dNTPs) by high-performance liquid chromatography, the choice of extracting agent and periodate oxidation procedure is important for accurate results. Different extraction methods were compared using either cold methanol or trichloroacetic acid for extraction of dNTPs from lymphoblastoid cells. The recoveries of the dNTPs varied with the different extraction methods. A modification of the periodate oxidation procedure for degrading ribonucleoside triphosphates to their bases was also compared with the original method. Both methods gave accurate results when trichloroacetic acid was used as extracting agent, but when methanol was used interfering peaks were present on the chromatogram when the original method was used. These peaks were absent when the modified periodate procedure was used.