Quantitative analysis of phosphorothioate oligonucleotides in biological fluids using fast anion-exchange chromatography.
Phosphorothioate oligonucleotides are potentially useful as anti-viral drugs. Classical DNA extraction methods are not as effective on short single-stranded DNA as with longer double-stranced chains. The classical method of phenol-chloroform extraction followed by ethanol precipitation is difficult to quantify, thus monitoring of the pharmacological disposition of these compounds is subject to error. A method has been devised and validated for extraction and analysis of modified oligonucleotides from biological fluids such as urine and serum based on protein kinase digestion and phenol-chloroform extraction. Due to the high native ultraviolet absorbance of the oligomers, detection limits in the low ppb range were obtained without derivatization.
Bourque
A J
AJ
Hybridon, Inc., Worcester, MA 01605.
Cohen
A S
AS
D002852:Chromatography, Ion Exchange / Q000379:methods*; D006801:Humans; D009841:Oligonucleotides / Q000032:analysis* / Q000097:blood / Q000652:urine; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D013873:Thionucleotides / Q000032:analysis* / Q000097:blood / Q000652:urine
D009841:Oligonucleotides; D013873:Thionucleotides