Determination of Steroid Hormone Residues in Fish Tissues Using Gel Permeation Chromatography and Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

A highly sensitive method was developed for the simultaneous determination of 8 steroid hormones in high-fat fish tissues using ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The 8 steroid hormones were extracted from the tissues with diethyl ether. Differing from other common purification methods, the extract solutions were cleaned by gel permeation chromatography (GPC) using ethyl acetate-cyclohexane solution (1:1, v/v) as the mobile phase. The separation of target compounds was carried out by a BEH C18 column and a gradient elution consisting of acetonitrile and 0.2% aqueous formic acid (v/v). The compounds were detected under the multiple reaction monitoring (MRM) mode and quantified with external standard method. This method was validated with respect to linearity, specificity, accuracy and precision. A linearity with correlation coefficient larger than 0.995 was achieved in the range of 0.5 to 50 ng mL−1. The average recoveries at the spiked levels of 1.0, 5.0, and 10.0 μg kg–1 varied between 81.7% and 90.8%, with the relative standard deviations (n=5) ranged from 3.50% to 10.0%. The limit of quantification (LOQ) for 8 steroid hormones ranged from 0.2 to 1.5 μg kg−1. It was concluded that this method can be successfully applied for the determination of 8 steroid hormones in complicated matrices including high-fat fish tissues.