Multi-Residue Determination of Eight Anabolic Steroids by GC-MS in Muscle Tissues from Pigs

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A gas chromatography-mass spectrometry(GC-MS)method to determine eight anabolic steroids(diethylstilbestrol,methyltestosterone,norethindrone,17α-ethynylestradiol,estradiol,6a-methyl-17α-hydroxy-progesterone,estradiol benzoate,and chlormadinone acetate)was developed.Muscle samples were extracted with liquid-liquid extraction and clean-up was performed in two steps,the extracts obtained were derivatized with heptafluorbutyric(HBF)anhydride and analyzed by GC-MS.In the above method,the linear scope was 2.5-50 μg kg-1.The range of the recoveries was 78.5-148% for diethylstilbestrol,70.8-109% for methyltestosterone,69.8-87.2% for norethindrone,67.7-120% for 17α-ethynylestradiol,82.8-103% for estradiol,70.3-99.2% for 6α-methyl-17α-hydroxy-progesterone,73.0-104% for estradiol benzoate,and 72.9-91.8% for chlormadinone acetate.The range of the coefficients of variation within batches was 0.4-12%;the range of the coefficients of variation between batches was 6.4-11%.The limit of detections and the detection capability were 0.99 and 3.30 μg kg-1 for diethylstilbestrol,1.05 and 3.50 μg kg-1 for methyltestosterone,1.19 and 3.97 μg kg-1 for norethindrone,0.94 and 3.13 μg kg-1 for 17α-ethynylestradiol,1.45 and 4.83 μg kg-1 for estradiol,1.56 and 5.20 μg kg-1 for 6α-methyl-17α-hydroxy-progesterone,1.92 and 6.40 μg kg-1 for estradiol benzoate,and 2.41 and 8.03 μg kg-1 for chlormadinone acetate,respectively.These results showed that the method was widely available,accurate,and sensitive.
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