Quick Determination of Three Hormones Residues in Beef and Kidney by HPLC-MS/MS

Trenbolone, testosterone and progesterone are steroid-assimilating hormone drugs, which are widely used in animal husbandry, especially in ruminants. Most hormones have potential carcinogenic effects. Long-term intake of assimilative hormones can lead to metabolic disorders, dysplasia or tumors. The residues of assimilating hormone drugs in liver, kidney and adipose tissue are higher, and the residues in muscle and plasma are lower. Therefore, the development of detection method for steroid assimilation hormone prototype drug in animal-derived food has significance. In this study, a high pressure liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was established for determination of the amount of hormones residues in beef and kidney products. The methyl tert-butyl ester solution was used for the extraction of steroid residues in beef and kidney products without solid phase extraction column. External standard method was used to quantify, so reducing pretreatment steps as much as possible, using high-efficiency extraction solvent, and then reducing sample loss, is the key factor to improve the detection limit of the method. The results showed that higher recovery could be obtained even without further purification and separation with tert-butyl methyl ether as extraction solvent, and the matrix interference had little effect. However, when the sample matrix was more complex (such as bovine kidney samples), especially in the case of more fat, direct detection without purification and separation will inevitably cause pollution to the instrument. Therefore, the method of centrifugal degreasing by freezing the sample at low temperature after rotary evaporation was used. This procedure not only plays an important role in purification, but also shortens the pretreatment time. The linearity was satisfactory with the correlation coefficient more than 0.996 7 in the range of 0.0-50.0 μg/L. Average recoveries for the steroid residues were in the range of 74%-83% with RSD below 10% in beef and kidney. The method is convenient, rapid and sensitive, which is suit for simultaneously determine three steroid residues in beef and kidney.