Determination of Nine Steroid Hormone Residues in Beef Samples by Gel Permeation Chromatography-Solid Phase Extraction-Rapid Resolution Liquid Chromatography-Mass Spectrometry/Mass Spectrometry

A gel permeation chromatography-solid phase extraction-rapid resolution liquid chromatography-tandem mass spectrometry (GPC-SPE-RRLC-MS/MS) method was developed for the determination of 9 steroid hormone residues in beef tissue. The beef samples were enzymatically digested with β-glucuronidase/arylsulfatase, and then extracted with tert-butyl methyl ether under ultrasonication incubation. Cleanup was carried out with GPC followed by a further HLB SPE. After C18 RRLC gradient elution separation with acetonitrile-0.1% aqueous formic acid as a mobile phase, the eluents were qualitatively and quantitatively determined under multireaction monitoring (MRM) scan type with tandem mass analyzer. The limit of quantification (LOQ) was 0.2-0.7 μg kg−1, and the calibration curves showed good linearity with correlation coefficients larger than 0.999. At the spiked levels of 0.2, 1.0 and 4.0 μg kg−1, the average recoveries were in the range of 81.4%-110%, and the relative standard deviations (RSDs) were in the range of 2.17%-9.82% (n = 7). The test results of real sample showed this method is sensitive and accurate. The proposed method was used for the sensitive and accurate determination of 9 steroid hormone residues in high-fat and complex samples.