Rapid Determination of Diarrhetic Shellfish Poisoning in Shellfish by Colorimetric Protein Phosphatase Inhibition Assay Based on Immobilization of Protein Phosphatase 2A in Sol-Gel

文献类型： 外文期刊

第一作者： Chen Jia-Qi

作者： Chen Jia-Qi;Mu Hai-Jin;Chen Jia-Qi;Wu Hai-Yan;Yao Lin;Zheng Guan-Chao;Guo Meng-Meng;Tan Zhi-Jun;Zhai Yu-Xiu;Chen Jia-Qi;Wu Hai-Yan;Yao Lin;Zheng Guan-Chao;Guo Meng-Meng;Tan Zhi-Jun;Zhai Yu-Xiu

作者机构：

关键词： Diarrhetic shellfish poisoning; Colorimetric protein phosphatase inhibition assay; Sol-gel; Shellfish; Rapid detection

期刊名称：CHINESE JOURNAL OF ANALYTICAL CHEMISTRY （ 影响因子：1.134； 五年影响因子：0.909 ）

ISSN： 0253-3820

年卷期： 2018 年 46 卷 8 期

页码：

收录情况： SCI

摘要： Based on the inhibition of protein phosphatase 2A (PP2A) by diarrhetic shellfish poisoning (DSP), mainly okadaic acid (OA) and its analogues (dinophysistoxins, DTXs), colorimetric protein phosphatase inhibition assay (PPIA) was built to detect DSP in shellfish in which 4-nitrophenyl phosphate (p-NPP) was used as the substrate. After hydrolyzed by PP2A, the product (4-nitrophenol, p-NP) was measured at 405 nm, then toxins in samples could be quantified according to the standard dose-effect curve developed with a series of toxin standard solutions. In order to overcome the instability of PP2A and preserve the enzymatic activity well, sol-gel, accompanied with hydroxyethyl-cellulose (HEC), was used to encapsulate and immobilize PP2A at the bottom of the microwells. Also, D-glucose was employed as the template or pore-forming agent to improve the sensitivity. The PP2A immobilized sol-gel exhibited a storage stability of 4 months at -18 degrees C. The limit of detection was 50 mu g OA eq./kg, the spiked recoveries for OA in shellfish samples were between 68. 8% and 119. 4%, and the linear range was from 50 to 800 mu g/kg. The PPIA with good recovery and reproducibility was simple and convenient. Samples and substrate solutions were added directly when used, demonstrating this proposed method could be used as an efficient analysis tool for rapid screening of DSP and be suited for regular monitoring program to control shellfish toxicity.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序