Validation of an analytical method for the determination of spiramycin, virginiamycin and tylosin in feeding-stuffs by thin-layer chromatography and bio-autography

摘要

An inter-laboratory validation was carried out to determine the performance characteristics of an analytical method based on thin-layer chromatography (TLC) coupled to microbiological detection (bio-autography) for screening feed samples for the presence of spiramycin, tylosin and virginiamycin. Twenty-four samples including blank samples and samples with concentrations of the target analytes ranging between 1 and 5 mg kg~(-1) (expressed in microbiological activity) were analysed by seven laboratories participating in the study. The required detection limit was 1 mg kg~(-1) (expressed in microbiological activity). For spiramycin, acceptable values for the sensitivity (at least 95%) indicating the rate of correct positive results were obtained for samples containing this substance at or above 2 mg kg~(-1), whereas at 1 mg kg~(-1), the sensitivity rate dropped to about 70%. Therefore, it was concluded that the detection limit was 2 mg kg~(-1). For tylosin and virginiamycin, acceptable values of the sensitivity were obtained for all concentrations including 1 mg kg~(-1). Therefore, the method fulfils the criterion regarding the required sensitivity at the target detection limit for tylosin and virginiamycin.