Efficient HPLC Separation of Diastereomers of Modified Oligodeoxynucleotides.

Library Number:
Part Number:
M. Cecilia Torres;Charles R. Iden
Fall ACS 2002; 224th ACS National Meeting; Boston August 18-22
Content Type:
Content Subtype:
DNA; Oligonucleotide; Diastereomers; BPDE
XTerra MS C18 N/A N/A N/A N/A
Both endogenous and exogenous chemical agents react with DNA and in many instances modify the structure of the nucleobase or the deoxyribose moiety. A single agent may produce products that are mixtures of stereoisomers, each of which may adopt a distinct chemical structure, have unique chemical stabilities and undergo reactions with specific DNA polymerases and repair enzymes. Modified oligodeoxynucleotides are used frequently to study the properties of these DNA adducts, and, ideally, each stereoisomer of a specific adduct should be isolated in order to elucidate differences in genotoxicity. We report several examples of the efficient separation of oligonucleotides containing diastereomers of critical DNA adducts using XTerra MS C18 columns (Waters Corporation). These include adducts from the reaction of acrolein, a reactive , alpha,beta-unsaturated aldehyde; oligodeoxynucleotides containing the N 2- benzo[alpha]pyrene diol epoxide (BPDE) adduct of deoxyguanosine; and diastereomers of thymine glycol in oligomers (23-mers). Excellent separation was achieved for these adducts resulting in HPLC fractions with oligomers containing a single stereoisomer.