Abstract: Using isotope internal standard dilution technique, a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method has been developed for the identification and quantitative determination of ciprofloxacin residue in the tissues of various fishes/shellfishes. The homogenized tissue sample added with ciprofloxacin-D8 and phosphate buffer solution (pH 7.0) was extracted with acetonitrile under ultrasonication, and degreased with hexane. After solid-phase extraction (SPE) was performed on an Oasis MAX cartridge, the sample was separated on a Cloversil-C18 column (150 mm×4.6 mm, 5 μm) by using the mobile phase consisting of CH3CN-0.05%CF3COOH (25∶75, v/v). The detection was carried out by LC-MS/MS using an electrospray ionization interface in multiple reaction monitoring (MRM) mode. The quantification using isotope-labelled internal standard was based on the peak area ratio of ciprofloxacin and deuterated internal standard in the MRM mode. The calibration curve was linear within the range of 0.1-50.0 μg/kg and the limit of quantification was 0.1 μg/kg (S/N≥10). The recovery was between 92.5% and 98.1%, and the relative standard deviation was less than 4.3%. The application of this method was further demonstrated by analyzing ten various real samples from local markets. The results show that this method is sensitive, accurate and suitable for the confirmative determination of ciprofloxacin residues.

Key words: ciprofloxacin, fish/shellfish tissues , isotope internal standard, liquid chromatography-tandem mass spectrometry (LC-MS/MS), solid-phase extraction （SPE）