Determination of oxytocin in biological samples by isocratic high-performance liquid chromatography with coulometric detection using C18 solid-phase extraction and polyclonal antibody-based immunoaffinity column purification.
A specific high-performance liquid chromatographic (HPLC) method is described for the reliable quantitation of oxytocin using culture media supernatants. The procedure employs solid-phase extraction, antibody-based immunoaffinity purification and isocratic HPLC with dual channel coulometric detection (ED). The lower limit of detection for this cyclic nonapeptide was 40 pg (40 fmol). Due to its relative simplicity, specificity and precision, the HPLC-ED of oxytocin is an accurate and attractive alternative to many existing quantitative methods.
Kukucka
M A
MA
Department of Biomedical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg 24061-0442.
Misra
H P
HP
D000595:Amino Acid Sequence; D000906:Antibodies; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003470:Culture Media; D004563:Electrochemistry; D008969:Molecular Sequence Data; D010121:Oxytocin / Q000032:analysis*; D015203:Reproducibility of Results
D000906:Antibodies; D003470:Culture Media; D010121:Oxytocin