The identification and quantitation of alkylated nucleobases by high-performance liquid chromatograghy with UV photodiode array detection

W. Xue, A. K. Samanta, Robert M Carlson

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

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. Quantitative spectral overlays of these compounds to methyl standard references from a spectral library and absorbance ratios at two maximal wavelengths (λ(max)) are found to be useful in categorizing the solutes. They can be grouped into classes of compounds originating from a specific nucleobase and classes of analogs having different alkyl substituents (e.g., methyl, ethyl, propyl, allyl, and benzyl) at the same position of the heterocycle. At a selected wavelength for alkylated nucleobases in the same class, the detector response factors are independent of the alkyl group (±10%). This 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.

Original languageEnglish (US)
Pages (from-to)477-481
Number of pages5
JournalJournal of Chromatographic Science
Volume28
Issue number9
DOIs
StatePublished - Jan 1 1990

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