Quantitative analysis of phosphorothioate oligonucleotides in biological fluids using direct injection fast anion-exchange chromatography and capillary gel electrophoresis.
The analysis of antisense phosphorothioate DNA (SODN) in human plasma via direct injection using anion-exchange high-performance liquid chromatography (AE-HPLC) is presented. The method relies on the ability to selectively extract phosphorothioate DNA from undigested serum, plasma and urine on anion-exchange resins. The automated HPLC method can analyze a sample every 5 min with a limit of detection of 50 ng/ml (ppb). The DNA was collected, desalted and analyzed by capillary gel electrophoresis. Due to the high resolving power of this technique, a qualitative assessment of enzymatic degradation of the antisense oligonucleotide can be made.
Bourque
A J
AJ
Hybridon, Worcester, MA 01605.
Cohen
A S
AS
D002852:Chromatography, Ion Exchange; D004247:DNA / Q000032:analysis; D004586:Electrophoresis; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000032:analysis* / Q000097:blood / Q000652:urine; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet; D013873:Thionucleotides / Q000032:analysis* / Q000097:blood / Q000652:urine
D016376:Oligonucleotides, Antisense; D012997:Solvents; D013873:Thionucleotides; D004247:DNA