Determination of steroid hormones in grey seal (Halichoerus grypus) blood plasma using convergence chromatography tandem mass spectrometry

A hybrid solid phase extraction (HybridSPE) protocol tailored to ultra-performance convergence chromatography tandem mass spectrometry (UPC2-MS/MS) was developed for the determination of 19 steroid hormones in grey seal (Halichoerus grypus) blood plasma. In this study, the protocol demonstrated acceptable absolute recoveries ranging from 33 to 90%. The chromatographic separation was carried out using a gradient elution program with a total run time of 5 min. For most target analytes, the method repeatability ranged from 1.9 to 24% and the method limits of quantification (mLOQs) ranged from 0.03 to 1.67 ng/mL. A total of 9 plasma samples were analysed to demonstrate the applicability of the developed method, and 13 steroid hormones were quantified in grey seal pup plasma. The most prevalent steroids: cortisol, cortisone, corticosterone, 11-deoxycortisol, progesterone and 17 alpha-hydroxyprogesterone were detected at concentrations in the range of 12.6-40.1, 7.10-24.2, 0.74-10.7, 1.06-5.72, 0.38-4.38 and <mLOQ - 1.01 ng/mL, respectively. To our knowledge, this is the first study to determine steroid hormones in the plasma of pinnipeds using convergence chromatography.