High-performance liquid chromatographic determination of spectinomycin in swine, calf and chicken plasma.

A rapid clean-up procedure based on ion-pair solid-phase extraction (SPE) for the high-performance liquid chromatographic (HPLC) determination of spectinomycin in swine, calf and chicken plasma at a limit of detection of 50 ng/ml is described. After dilution with water and adjustment of the pH to approximately 5.6, the plasma is applied to a high-hydrophobic C18 SPE column treated with sodium dioctylsulphosuccinate. Spectinomycin is eluted with methanol and derivatized with 2-naphthalene sulphonyl chloride prior to chromatography. The HPLC set-up consists of a dual-column system using two Chromspher silica columns and dichloromethane-acetonitrile-ethyl acetate-acetic acid, in different ratios, as mobile phases. Detection is performed at 250 nm. Quantification is carried out using external standards prepared in blank cleaned plasma. Mean recoveries were 83 +/- 3% (n = 5), 93 +/- 6% (n = 5) and 92 +/- 6% (n = 6) for swine, calf and chicken plasma, respectively, at the 0.1 microgram/ml level.