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IDENTIFICATION AND QUANTITATION OF ALKYLATED NUCLEOBASIS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH UV PHOTODIODE ARRAY DETECTION
Citation:
Xue, W., A. Samanta, AND R. Carlson. IDENTIFICATION AND QUANTITATION OF ALKYLATED NUCLEOBASIS BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH UV PHOTODIODE ARRAY DETECTION. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-90/252.
Description:
The application of UV diode array detection in high-performance liquid chromatographic (HPLC) identification and quantitation of several classes of synthetic and commercially available alkylated nucleobases is investigated. uantitative spectral overlays of these compounds to methyl standard references from a spectral library and absorbance ratios at two maximal wavelengths are found to be useful in categorizing the solutes. hey can be grouped into classes of analogs having different alkyl substituents (e.g., methyl, ethyl, propyl, allyl, and benzyl) at the same position of the heterocycle. t a selected wavelength for alkylated nucleobases in the same class, the detector response factors are independent of the alkyl group (+ 10%). his technique provides a practical means for both qualitative and quantitative analysis of product distribution of DNA base alkylation by using only readily obtainable methylated derivatives as the reference standards.