The obtained spectrums were analyzed by Thermo Proteome Discoverer 1.4 software and then ANIQLVEK with high sensitivity was selected as the 20-Hydroxyecdysone quantitative signature peptide. that the developed method could be used for the determination of TM in different kinds of food samples. Table 6 Results of real samples. thead th valign=”top” align=”left” rowspan=”1″ colspan=”1″ Sample /th th valign=”top” align=”center” rowspan=”1″ colspan=”1″ TM (g/g) /th /thead Penaeus vannamei3,291China shrimp3,730Fresh shrimp slicesNDFresh shrimp stripsNDShrimp ballsNDLobster steakNDLobster stickNDCrab king stickNDCrab chopsNDFish ballNDCuttle ballNDDragon prawn ballND Open in a separate window The Advantages of the Method To compare relevant papers in the field, immunoaffinity purification was used in the procedure of sample pretreatment for first time. TM extraction, purification, antibody preparation, and the preparation of immunoaffinity column 20-Hydroxyecdysone were finished by members of the research group. Signature peptides and isotope-labeled internal standard were used in the quantitative analysis. The method has the advantages of simple pretreatment, less interference, high specificity, and perfect accuracy and precision. Conclusion In this experiment, TM was extracted and purified, antibody of TM was made, and immunoaffinity column was filled. After simple extraction, the samples were purified by immunoaffinity column and then digested by trypsin. A new liquid chromatographyCtandem mass spectrometry method at the peptide level was developed to determine TM, NIQLVEK (AK-8) was confirmed as the quantitative peptide and synthesized for the further 20-Hydroxyecdysone process, and isotope-labeled internal standard was used in the quantitative analysis. The specificity, linearity, sensitivity, matrix effect, accuracy, and precision of the method were investigated, and Mouse monoclonal to Epha10 the developed method has been successfully used for the detection of TM in various food samples. Data Availability Statement The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation. Ethics 20-Hydroxyecdysone Statement The animal study was reviewed and approved by Animal Ethical and Welfare Review Committee of Hebei Food Inspection and Research Institute. Author Contributions YZ and YW conceived and designed the experiments. SF, JM, QL, CL, and JZ performed the experiments. YW analyzed the data. WZ and LW wrote the original draft. All authors have read and approved the manuscript. Funding This work was supported by Hebei Province High-Level Talent Funding Program (Project No. A201901008), Science and Technology Project of State Administration for Market Regulation (Project No. 2021MK023), and Research Project of Hebei Administration for Market Regulation (Project No. 2020ZD12). Conflict of Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Publisher’s Note All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher..
