The identification and quantitation of alkylated nucleobases by high-performance liquid chromatography with UV photodiode array detection.
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.
Xue
W L
WL
Department of Chemistry, University of Minnesota, Duluth 55812.
Samanta
A K
AK
Carlson
R M
RM
D000225:Adenine / Q000031:analogs & derivatives* / Q000032:analysis; D000478:Alkylation; D002851:Chromatography, High Pressure Liquid; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis; D012015:Reference Standards; D013056:Spectrophotometry, Ultraviolet; D013941:Thymine / Q000031:analogs & derivatives* / Q000032:analysis; D014498:Uracil / Q000031:analogs & derivatives* / Q000032:analysis
D014498:Uracil; D006147:Guanine; D000225:Adenine; D013941:Thymine