Development and validation of a confirmatory HPLC method for simultaneous determination of Sudan dyes in animal...

A simple and sensitive high-performance liquid chromatography (HPLC) analytical method for the simultaneous determination of six Sudan dyes (Sudan Red G, Sudan I, Sudan II, Sudan III, Sudan Red 7B, Sudan IV) in animal tissues and eggs was developed. Samples were extracted with acetonitrile followed by a cleanup using a C(18) solid-phase extraction column. Chromatographic separation was achieved on a Zorbax SB-C(18) column under gradient conditions. The analytes were detected at 510 nm by HPLC with diode array detection. Specificity, decision limit (CCalpha), detection capacity (CCbeta), accuracy, and precision were determined during validation process. Recoveries for six Sudan dyes from three animal tissues and eggs were 77.2-98.0% with excellent relative standard deviations. CCalpha and CCbeta were in the range of 7.7-9.0 microg/kg and 9.1-10.3 microg/kg, respectively. The limits of quantitations were between 12.8 microg/kg and 15.0 microg/kg.