An in vitro stability study of unmodified and modified antisense oligonucleotides in human serum was performed with a previously developed capillary electrophoretic method using either micellar solution or entangled polymer solution depending on the oligonucleotide length to be separated. A method has been devised and validated for the extraction of oligonucleotides from serum using anion-exchange centrifugal filter units. The extracted samples were desalted by a drop dialysis method. The serum half-lives and the degradation patterns of unmodified and modified oligonucleotides are compared. The modified oligonucleotide used in this study is protected from exonuclease activity present in human serum by terminal 1,3-propanediol modification.