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渔业研究 ›› 2022, Vol. 44 ›› Issue (2): 146-153.DOI: 10.14012/j.cnki.fjsc.2022.02.005

• 论文与报告 • 上一篇    下一篇

ELISA法测定水产苗种中孔雀石绿及其代谢物的残留量

梁娟张倩玉魏海英于盟盟牟丹丹王磊   

  1. 日照市海洋与渔业研究院,山东 日照 276826
  • 收稿日期:2021-12-14 修回日期:2022-01-17 出版日期:2022-04-25 发布日期:2022-04-25
  • 通讯作者: 于盟盟(1975-),男,工程师,本科,从事水产养殖与增殖和水产品质量安全检测. E-mail:157017844@qq.com
  • 作者简介:梁 娟(1975-),女,高级工程师,本科,从事水产养殖与增殖和水产品质量安全检测. E-mail:liangjuan75@126.com
  • 基金资助:
    国家虾蟹产业技术体系建设任务书(CARS-48);山东省现代农业产技术体系藻类创新团队建设专项资金(SDAIT-26);山东省现代农业刺参产业技术体系任务书(SDAIT-22-12);山东省现代农业鱼类产业技术体系任务书(SDAIT-12-08)

Determination of malachite green and its metabolite residues in aquatic seedlings by ELISA

  • Received:2021-12-14 Revised:2022-01-17 Online:2022-04-25 Published:2022-04-25

摘要: 本文建立了水产苗种中孔雀石绿及其代谢物隐色孔雀石绿残留量的酶联免疫(ELISA)法。采用乙腈作为提取剂、超声提取替代涡旋振荡提取、纯水对正己烷进行饱和、红色记号笔对离心管进行标记等措施对前处理过程进行优化。结果表明:应用ELISA法测定孔雀石绿残留量,在0.050 0~4.50 ng·mL-1范围内呈现良好的线性关系,相关系数R2=0.997 0,方法检出限为0.075 0 μg·kg-1。在基质样品添加浓度为0.075 0、0.150、0.750 μg·kg-1的加标水平下,中国对虾、三疣梭子蟹、刺参、牙鲆等苗种的平均回收率为79.7%~104.5%,精密度(RSD)为3.3%~7.9%。试验结果表明,该法灵敏度高、特异性强、快速简便,适用于不同基质水产苗种中孔雀石绿及其代谢物隐色孔雀石绿残留的快速批量检测,能及时为渔业质量安全监管部门的执法工作提供强有力的技术支撑。

关键词: 水产苗种, 酶联免疫法, 孔雀石绿, 隐色孔雀石绿, 残留量

Abstract: An enzyme-linked immunosorbent assay (ELISA) for detecting the residue of malachite green (MG) and its metabolite leucomalachite green (LMG) in aquatic seedlings was established in the present study. The pretreatment process was optimized by using acetonitrile as the extractant, ultrasonic extraction instead of vortex oscillation extraction, saturated n-hexane with pure water, and marking the centrifugal tube with red marker. The results showed that MG content had good linear in the range of 0.050 0~4.50 ng.mL-1, which the correlation coefficients was R2=0.997 0, and the detection limit was 0.075 0 μg·kg-1. The average recoveries ranged from 79.7%~104.5% at the spiked level of 0.075 0, 0.150 and 0.750 μg·kg-1 in the seedlings of Penaeus chinensis, Portunus trituberculatus, Stichopus japonicus and Paralichthys olivaceus. The relative standard deviations (RSD) was 3.3%~7.9%. The results showed that this method was accurate, rapid and cheap, which would be suitable for periodic spot check of MG & LMG residues in different aquatic seedlings substrate.

Key words: aquatic seedlings, ELISA, malachite green, leucomalachite green, residue

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