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Updated: Aug 4, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
High-performance liquid chromatographic determination of spectinomycin in swine, calf and chicken plasma using
N Haagsma1, P Scherpenisse, R J Simmonds
1Department of the Science of Food of Animal Origin, Faculty of Veterinary Medicine, University of Utrecht, Netherlands.
Abstract:
An HPLC method for the determination of spectinomycin in swine, calf and chicken plasma at 0.1 microgram/ml or higher is described. The clean-up is based upon ion-pair solid-phase extraction on a High Hydrophobic C18 column treated with sodium dioctyl sulfosuccinate. After elution with methanol, spectinomycin is chromatographed on a Spherisorb SCX column using 0.1 M sodium sulphate solution (pH 2.6)-acetonitrile (80:20, v/v) as mobile phase. Fluorescence detection is at an excitation wavelength of 340 nm and an emission wavelength of 460 nm after post-column oxidation with sodium hypochlorite followed by derivatization with o-phthaldialdehyde. Mean recoveries were 99 +/- 2% (n = 6), 99 +/- 2% (n = 7) and 104 +/- 2% (n = 6) for swine, calf and chicken plasma, respectively, at the 0.1 microgram/ml level.
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