Determination of oxytocin in biological samples by isocratichigh-performance liquid chromatography with coulometric detection using C18solid-phase extraction and polyclonal antibody-based immunoaffinity columnpurification

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

by Mark A. Kukucka and Hara P. Misra
Department of Biomedical Sciences
Virginia-Maryland Regional College of Veterinary Medicine
Virginia Polytechnic Institute & State University, Blacksburg 24061


ABSTRACT

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.


CITATION
Kukucka, Mark A. and Hara P. Misra. 1994. 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. J Chromatogr B
Biomed
Appl 653(2):139-45.


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