A combined solid phase extraction/capillary gel electrophoresis method for the determination of phosphorothioate oligodeoxynucleotides in biological fluids, tissues and feces.
A novel biological sample clean-up procedure has been developed for the determination of phosphorothioate oligodeoxynucleotides (PS-ODNs) and derived metabolites in biological fluids (plasma, urine and bile) and in tissues and feces from mice and rats. This method uses a one-step C18 solid-phase extraction (SPE) for biological matrix removal, and it uses capillary gel electrophoresis (CGE) for analyte detection. The assay is specific, and its linearity is superb (r>0.99) for IV-AS (a 13-mer PS-ODN) and PS19 (a 19-mer PS-ODN) in a variety of biological matrices. For both IV-AS and PS19, the precision, accuracy and absolute recovery values were found to be <20%, +/-20% and 80-120%, respectively. The LODs of IV-AS and PS19 were 0.6 mg/l for plasma, 0.8 mg/l for rat urine and bile, 6 microg/g for rat tissues, and 10 microg/g for rat feces, with a signal-to-noise ratio of 3 (S/N=3). This method has been successfully applied to the analysis and quantitation of PS-ODNs in various biological samples arising from preclinical pharmacokinetic studies.
Wu
Li-Xia
LX
Beijing Institute of Radiation Medicine, Beijing 100850, PR China.
Lu
Dan-Dan
DD
Zhou
Zhe
Z
Zhang
Hong-Yan
HY
Zhang
Yu-Lin
YL
Wang
Sheng-Qi
SQ
D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D001483:Base Sequence; D001826:Body Fluids / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005243:Feces / Q000737:chemistry*; D051379:Mice; D009838:Oligodeoxyribonucleotides / Q000032:analysis* / Q000493:pharmacokinetics; D009943:Organophosphorus Compounds / Q000032:analysis* / Q000493:pharmacokinetics; D051381:Rats; D012015:Reference Standards; D012680:Sensitivity and Specificity; D012965:Sodium Chloride / Q000737:chemistry; D013873:Thionucleotides / Q000032:analysis* / Q000493:pharmacokinetics; D014018:Tissue Distribution
D000097:Acetonitriles; D009838:Oligodeoxyribonucleotides; D009943:Organophosphorus Compounds; D013873:Thionucleotides; D012965:Sodium Chloride; C032159:acetonitrile