A method for rapid screening and identification of 30 protein assimilation hormones and glucocorticoids in fish was established using ultra-high performance liquid chromatography coupled to quadrupole/orbitrap high-resolution mass spectrometry (UPLC-Q/Orbitrap HRMS). The samples were extracted with 80% acetonitrile aqueous solution (containing 0.2% formic acid), centrifuged, purified by solid phase extraction (SPE) on Oasis PRiME HLB column, and dried with nitrogen blow. The residue was re-dissolved. The chromatographic separation was performed on a Waters Acquity BEH C₁₈ (2.1 mm × 100 mm, 1.7 μm) column through gradient elution using a mobile phase consisting of 20 mmol/L ammonium acetate aqueous solution containing 0.1% formic acid (A) and acetonitrile (B). The target analytes were detected in the positive and negative ion mode using a heated electrospray ionization (HESI) source through primary full scan/data-dependent secondary scan (full MS/dd-MS²) monitoring, and quantitated by matrix-matched external standard method. The results showed that good linearity was observed for the 30 hormones in the concentration range of 0.5–100 ng/mL with correlation coefficients (r) greater than 0.9950. The limits of detection were between 0.2 and 1.0 μg/kg, and the limits of quantification were between 0.5 and 2.0 μg/kg. The recoveries were 69.7%–103.2% with relative standard deviations (RSDs) of 2.3%–9.4% at three spiked levels for four different fishes (turbot, mandarin fish, mullet and grass carp). The proposed method is efficient, accurate and reliable, and is suitable for the rapid multi-objective screening and quantitative analysis of protein assimilation hormones and glucocorticoids in fish.

Monitoring of food chemical hazards (FCHs) is crucial to ensure food quality and safety. Non-targeted screening (NTS) is an effective technique to identify a wide range of potential and unknown FCHs using high performance liquid chromatography/high performance gel chromatography-high resolution mass spectrometry (HPLC/HPGC-HRMS) platforms with strong separation capability, high mass resolution and high mass accuracy. However, the development and application of NTS methods are still challenging due to the wide variety of FCHs, the large amount of acquired data, and the cumbersome analysis process. Selecting an appropriate data acquisition mode and an effective non-targeted data analysis strategy are essential to successfully identify potential and unknown FCHs. This paper reviews the data acquisition modes, analysis processes and strategies involved in NTS and the application of NTS for FCHs screening in the past five years, in order to promote the development of NTS methods for FCHs and to provide support for the early warning of risk factors in foods and food safety monitoring.