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Validation and quantification of a UPLC-MS/MS method for the simultaneous determination of quinocetone and its main metabolites (3-methylquinoxaline-2-carboxylic acid and dedioxoquinenone) in aquatic products

Tian Xiuhui, Han Dianfeng, Cui Yanmei, Ren Lihua, Jiang Fang, Huang Hui, Gong Xianghong, Xue Jinglin, Li Jiawei, Liu Huihui, Xu Yingjiang, Luo Xiaojun, Liu Xiaojing, Zhang Xiuzhen

Acta Chromatographica

2022

Vol. 34, no. 4

444--452

A sensitive and validated method for determining quinocetone and its main metabolites (3-methylquinoxaline-2-carboxylic acid and dedioxoquinenone) was established in aquatic products using ultra-high-performance liquid chromatography-tandem spectrometry (UHPLC-MS/MS). Samples were extracted with 2.0 mol L⁻¹ hydrochloric acid, then purified on MAX columns. After extraction and purification, the supernatant was evaporated to dry nearly under a gentle stream of nitrogen at 40 °C. Formic acid-acetonitrile-water (0.1/30/70, v/v/v) was adjusted to 1.00 mL final volume. An aliquot (10 μL) was injected into the C18 column for separation with the mobile phase of acetonitrile and 0.5% formic acid in water at 0.25 mL min⁻¹. Calibration curves were linear ranged from 10.00 ng mL⁻¹ to 200.0 ng mL⁻¹ for quinocetone and 3-methylquinoxaline-2-carboxylic acid, and 20.00 ng mL⁻¹ to 400.0 ng mL⁻¹ for dedioxoquinenone. Mean recoveries were 70%–89%, 73%–83% and 72%–84%, respectively. The limit of detection (LOD) was 1.00 μg kg⁻¹, 1.00 μg kg⁻¹ and 2.00 μg kg⁻¹, and quantification (LOQ) were 2.00 μg kg⁻¹, 2.00 μg kg⁻¹ and 4.00 μg kg⁻¹ for quinocetone, 3-methylquinoxaline-2-carboxylic acid, and dedioxoquinenone. Based on the method above, the analytes were determined in Apostichopus japonicus, three fishes (including Ctenopharyngodon idellus, Crucian carp and Oreochromis mossambicus), Penaeus vannamei, Penaeus chinensis, and Chlamys farreri. The method shows good sensitivity, linearity, precision, and accuracy. In short, the proposed method was reliable for the determination of quinocetone, 3-methylquinoxaline-2-carboxylic acid, and dedioxoquinenone in aquatic products.