GC-MS method development and validation for anabolic steroids in feed samples

• Published in: Journal of separation science Vol. 31; no. 4; pp. 727 - 734
• Main Authors: Muñiz-Valencia, Roberto, Ceballos-Magaña, Silvia G, Gonzalo-Lumbreras, Raquel, Santos-Montes, Ana, Izquierdo-Hornillos, Roberto
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley-VCH Verlag 01.03.2008; WILEY-VCH Verlag; WILEY‐VCH Verlag; Wiley
• Subjects: Anabolic Agents - analysis; Anabolic steroids; Animal feed; Animal Feed - analysis; Animal productions; Animals; Biological and medical sciences; Derivatization; Feed and pet food industries; Food industries; Fundamental and applied biological sciences. Psychology; Gas Chromatography-Mass Spectrometry - methods; Growth promoting agents; Steroids - analysis; Swine; Terrestrial animal productions; Testosterone Congeners - analysis; Vertebrates; Steroid; Chemical analysis; Silylation; Derivatization; Solvent extraction; Veterinary medicine; Growth promoting agents; Sample preparation; Ungulata; Feed; Quantitative analysis; Validation; Farming animal; Liquid liquid extraction; Coupled method; Young animal; Animal feed; Pig; Programmed temperature; Gas chromatography; Vertebrata; Mammalia; Anabolic steroids; Artiodactyla; Anabolic agent; Mass spectrometry
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.200700513

A GC-MS method for the determination of AAS used as growth promoting agents using SIM in piglet feed samples has been developed and validated, using testosterone as internal standard. The formation of volatile steroid derivatives was carried out by derivatization with N-methyl-N-(trimethylsilyl)trifluoroacetamide. The optimum separation was achieved using a Zebron ZB-5 column under a gradient temperature elution, allowing the separation of steroids in 18 min. The required sample treatment process was discussed. A leaching using ACN, saponification using a binary NaOH/MgCl₂ solution, and LLE using ethyl acetate were finally selected. Method validation has been carried out according to the Commission Decision 2002/657/EC criteria established for quantitative confirmatory methods. The extraction efficiencies, CCα and CCβ for these compounds were in the ranges 78-98%, 10-21 and 18-35 μg/kg, respectively. The repeatability and the within-laboratory reproducibility at 1, 1.5, and 2 CCβ concentration levels were smaller than 8.2, 7.5, and 5.8% and 12.2, 9.5, and 7.5%, respectively. Accuracy was in the 99-103% range. The robustness was evaluated using the Youden robustness test. The proposed method was applied to the analysis of steroids spiked in different kinds of animal feed samples with satisfactory results.