Xue W L, Samanta A K, Carlson R M
Department of Chemistry, University of Minnesota, Duluth 55812.
J Chromatogr Sci. 1990 Sep;28(9):477-81. doi: 10.1093/chromsci/28.9.477.
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 (lambda 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.
研究了紫外二极管阵列检测在高效液相色谱（HPLC）中对几类合成的和市售的烷基化核碱基进行鉴定和定量分析中的应用。发现这些化合物与光谱库中甲基标准参比物的定量光谱叠加以及在两个最大波长（λmax）处的吸光度比，对于溶质的分类很有用。它们可以分为源自特定核碱基的化合物类别以及在杂环相同位置具有不同烷基取代基（例如甲基、乙基、丙基、烯丙基和苄基）的类似物类别。在同一类烷基化核碱基的选定波长下，检测器响应因子与烷基无关（±10%）。该技术仅使用易于获得的甲基化衍生物作为参比标准，为DNA碱基烷基化产物分布的定性和定量分析提供了一种实用方法。
