建立高效液相色谱串联三重四极杆方法（HPLC-MS/MS），对喹烯酮在刺参体壁中的未知代谢物进行定性检测和结构解析。单次投喂刺参含喹烯酮20 mg/kg的饲料，24 h后转入清洁海水中72 h，实验期间定时取样去除内脏进行分析。本研究采用乙腈直接提取浓缩至干后定容检测和盐酸酸解后过SPE柱富集净化检测两种前处理方式对刺参体壁中的喹烯酮和可能代谢物进行提取和净化。采用ACQUITYTM UPLC ? BEH C18（2.1 mm ? 50 mm，1.7 μm）色谱柱，乙腈（0.1% HCOOH）和水（0.1% HCOOH）为流动相，对未知代谢物样品进行梯度洗脱分离，电喷雾离子源（ESI）正离子，Fullscan-Data Dependant scan （Fullscan-DDA）扫描方法进行检测。通过检测组分与已知喹烯酮和代谢物标准品的一级母离子和二级碎片离子对比，在刺参体壁中确认了已知代谢物MQCA（3-甲基喹恶啉-2羧酸），并首次发现了另4种可能代谢物。

A high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method was established to conduct qualitative analysis and structural elucidation of the unknown quinocetone metabolites in the body wall of Apostichopus japonicus. Apostichopus japonicus were exposed to the one-time feed containing 20 mg/kg of quinocetone for 24 h, and then were transferred to clean seawater for 72 h. Sea cucumber samples were collected regularly during the experiment for analysis after removal of the viscera. In this study, acetonitrile was used for direct extraction, the extract was dried, and the dried sample was reconstituted in a volumetric flask. After hydrochloric acid hydrolysis, sample enrichment and purification were performed on a solid-phase extraction (SPE) cartridge to detect the quinocetone in the Apostichopus japonicus body wall due to the two pre-treatments as well as to extract and purify the possible metabolites. The metabolites were separated on an ACQUITYTM ? BEH C18（2.1 mm?50 mm, 1.7 μm）column by gradient elution with acetonitrile-water (containing 0.1% formic acid) as the mobile phase, and were detected using full-scan-data-dependent scan (Fullscan-DDA) by positive ion electrospray ionization (ESI+). By comparing the MS1 and MS2 ions of the test sample with those of quinocetone and other metabolite standards, the metabolite MQCA (3-methyl-quinoxaline-2-carboxylic acid) was identified in Apostichopus japonicus body wall, along with other four possible, metabolites for the first time.

山东省科学技术发展计划项目（2012GHY11517）；山东省水生动物营养与饲料泰山学者岗位资助项目