Synthetic and natural hormones are used in the production and intensive breeding of animals as growth factors and reproductive regulators. However, the illegal and inappropriate use of these substances increases the risk of introducing residues into the food chain. Also because of their carcinogenic and teratogenic effects, the European Union has banned the use of these substances since 1985. For this reason, the development of sensitive and reliable analytical methods for the monitoring of their residues in food has become a necessity. A liquid chromatography analysis method coupled to mass spectrometry and based on Quechers extraction has been developed to analyse thirteen synthetic and natural hormones in muscles. In order to study the reliability of this method, its validation has been carried out in different bovine matrices (liver, kidney, bile, and hair) according to European Decision 2002/657 / EC. The method demonstrates good linearity (R2> 0.99) as well as accuracy with coefficients of variation for repeatability and reproducibility less than 23%. The values of CCα and CCβ were determined for each analyte indicating values ranging from 0.13 to 0.86 μg.kg-1 and 0.25 to 1.72 μg.kg-1 respectively for most of the analytes. Higher values were obtained for 17β-estradiol, estriol, 17α-ethinyl-estradiol ranging from 1.73 to 13.95 μg.kg-1 for CCα and from 3.47 to 23.87 μg.kg-1 for CCβ. The recovery rate in the different matrices (liver, kidney, bile, and hair) varies from 51.5 to 107 %. The matrix effect of the method was evaluated, indicating significant suppression values for the liver and the kidney, which varied respectively from -45 to -15.5 % and -35 to - 2.5%. In the same way, the other two matrices hair and bile show smaller matrix effects than the others. Finally, this method has been successfully applied to detect anabolic hormones in nighty one samples (muscle, liver, kidney, bile) available from different local butchers. Progesterone was found in 41 samples at 0.11 to 11.7 µg.kg−1 otherwise testosterone was observed in 43 samples ranging from 0.5 to 9.72 µg.kg−1.
VALIDATION OF CONFIRMATORY MULTI-RESIDUE METHOD FOR THE DETERMINATION OF THIRTEEN HORMONES RESIDUES IN SEVERAL BOVINES MATRICES USING HPLC-MS/MS
Fadl Moussa;Nicola Bernabo;
2019-01-01
Abstract
Synthetic and natural hormones are used in the production and intensive breeding of animals as growth factors and reproductive regulators. However, the illegal and inappropriate use of these substances increases the risk of introducing residues into the food chain. Also because of their carcinogenic and teratogenic effects, the European Union has banned the use of these substances since 1985. For this reason, the development of sensitive and reliable analytical methods for the monitoring of their residues in food has become a necessity. A liquid chromatography analysis method coupled to mass spectrometry and based on Quechers extraction has been developed to analyse thirteen synthetic and natural hormones in muscles. In order to study the reliability of this method, its validation has been carried out in different bovine matrices (liver, kidney, bile, and hair) according to European Decision 2002/657 / EC. The method demonstrates good linearity (R2> 0.99) as well as accuracy with coefficients of variation for repeatability and reproducibility less than 23%. The values of CCα and CCβ were determined for each analyte indicating values ranging from 0.13 to 0.86 μg.kg-1 and 0.25 to 1.72 μg.kg-1 respectively for most of the analytes. Higher values were obtained for 17β-estradiol, estriol, 17α-ethinyl-estradiol ranging from 1.73 to 13.95 μg.kg-1 for CCα and from 3.47 to 23.87 μg.kg-1 for CCβ. The recovery rate in the different matrices (liver, kidney, bile, and hair) varies from 51.5 to 107 %. The matrix effect of the method was evaluated, indicating significant suppression values for the liver and the kidney, which varied respectively from -45 to -15.5 % and -35 to - 2.5%. In the same way, the other two matrices hair and bile show smaller matrix effects than the others. Finally, this method has been successfully applied to detect anabolic hormones in nighty one samples (muscle, liver, kidney, bile) available from different local butchers. Progesterone was found in 41 samples at 0.11 to 11.7 µg.kg−1 otherwise testosterone was observed in 43 samples ranging from 0.5 to 9.72 µg.kg−1.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
