Development of an HPLC-DAD Method for the Determination of Five Sulfonamides in Shrimps and Validation According to the European Decision 657/2002/EC

Abstract

A method for the determination of five sulfonamide drugs: sulfadiazine (SDZ), sulfamethazine (SMTH), sulfisoxazole (SIX), sulfadimethoxine (SDMX), sulfaquinoxaline (SQX) in shrimps was developed using a high-pressure liquid chromatography–diode array detection (HPLC–DAD) instrumentation. The drugs were extracted from the samples using a solid-liquid extraction, with a mixture of acetonitrile-methanol and aqueous solution of acetic acid. The separation was achieved with LiChroCART-LiChrospher® 100 RP-18 (5 μm, 250 × 4 mm), and the mobile phase consisted of (A) aqueous solution of acetic acid 0.1% (65%) and (B) mixture of ACN-MeOH 50:50 v/v (35%), delivered at a flow rate of 0.8 mL/min isocratically. A DAD detector was used for the detection of the sulfonamides at 265 nm. The method was validated according to the 657/2002/EC decision of the European Union. The recoveries for within-day repeatability were 90.2–109.0% and for between-day precision 88.6–108.4%. The limit of quantitation for all five suflonamides was 50 μg/kg, and the limit of detection was 15 μg/kg.