Determination of Steroids in Porcine Plasma by High Performance Liquid Chromatography-Triple Quadrupole-Tandem Mass Spectrometry

KeyWord:steroids  hormone  high performance liquid chromatography tandem mass spectrometry(HPLC-MS/MS)  porcine plasma

Abstract:

      A method was developed for the determination of 33 kinds of steroid hormones£¨including androgens,estrogens,progestogens,corticosteroids and steroids-like chemicals£© in porcine plasma.The analytes in samples were extracted with methyl tert-butyl ether by vortex oscillation.The separation of analytes was achieved by gradient elution on a Xselect-C18 column£¨3.5 ¦Ìm,2.1 mm¡Á100 mm£© under positive mode and an Extend-C18 column£¨3.5 ¦Ìm,2.1 mm¡Á100 mm£© under negative mode.All steroids were detected by high performance liquid chromatography combined with tandem mass spectrometry£¨HPLC-MS/MS£© and quantified by the external standard method.The calibration curves of 33 steroids were linear in the certain concentration range with correlation coefficients£¨r£© more than 0.99.The average recoveries of steroids at three spiked levels ranged from 50.9% to 119.5% with relative standard deviations£¨RSDs£© of 2.3%-9.8%.The limits of quantitation£¨LOQ,S/N¡Ý10£© were in the range of 0.05-40.00 ng/mL.The method was used for the determination of steroids in real samples collected from the large scaled livestock farms in Beijing and 10 kinds of steroids£¨including testosterone,progesterone,cortisone,androstenone,hydrocortisone,4-androstenedione,corticosterone,cortexolone,21-hydroxyprogesterone and 17¦Á-hydroxyprogesteron£© were detected.The results showed that the method was simple,sensitive and reproducible,and could be applied in the determination of steroids in porcine plasma.