Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
40, Number 9, October 2002, pp. 515-518
Determination of Bovine Blood Oleandrin by High-Performance Liquid
Chromatography and Postcolumn Derivatization
Kimio Hamada[1],[2], Asako Iwamoto[1],[3], Shigeru Miyazaki*,[1],
Noriko Yamanaka[1], and Keerthi Siri Guruge[1]
[1]Toxico-Biochemistry Section, National Institute of Animal Health, Kannondai
3-1-5, Tsukuba, Ibaraki 305-0856, Japan;
[2]Kumamoto Central Animal Hygiene Service Center, Shizume 1666, Johnan, Shimomashiki,
Kumamoto 861-4215, Japan; and
[3]Himeji Animal Hygiene Service Center, Tadera-higashi 2-10-16, Himeji, Hyogo
670-0081, Japan
A high-performance liquid chromatographic procedure with a postcolumn
fluorescence derivatization is developed for the analysis of oleandrin in bovine
blood. Oleandrin is separated by an octadecylsilane-bonded column with a mobile
phase containing dehydroascorbic acid. The effluent of the column is mixed with
concentrated hydrochloric acid and passed through poly(tetrafluoroethylene)
tubing maintained at 70°C. The resultant fluorophores are detected at 465
nm with excitation at 348 nm. Simple solid-phase extraction using Sep-Pak tC2
is effective for sample purification. We found the minimal detectable quantity
of oleandrin in plasma to be 1.5 ng/mL at a signal-to-noise ratio of 3:1.
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