A Method for Determining Ten Alkaloids in Feed: Rapid and Accurate Quantification Based on Ultra‐High Performance Liquid Chromatography‐Tandem Mass Spectrometry

Abstract

In this study, ultra‐high performance liquid chromatography‐tandem mass spectrometry (UHPLC‐MS/MS) was employed for the rapid detection and analysis of 10 alkaloids in feed, such as morphine, caffeine, and sanguinarine. The methodology involved sample extraction using methanol, purification with an amino column (NH2), separation via a Hypersil GOLD C18 column (100 × 2.1 mm, 1.9 µm), and elution using a methanol‐0.1% formic acid aqueous solution as the mobile phase, with a flow rate of 0.3 mL min−1 and a column temperature of 40°C. Ten compounds showed a good linear range within the range of 0.01–5 ng mL−1, with a correlation coefficient (R2) exceeding 0.99. The limit of detection (LOD) ranged from 0.002 to 0.01 mg kg−1. Peak areas demonstrated excellent linearity across various concentrations. Furthermore, the study determined the LOD and LOQ for the 10 alkaloids. The recovery rate for different feed types ranged from 95.1% to 116.8%, with a relative deviation (RSDn = 6) of 0.86% to 1.31%. Overall, this method offers good reproducibility, high sensitivity, and stability, making it suitable for the detection and analysis of alkaloids in feed.