Genotyping of Toxic Pufferfish Based on Specific PCR-RFLP Products As Determined by Liquid Chromatography/Quadrupole-Orbitrap Hybrid Mass Spectrometry

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• 作者：Hajime Miyaguchi ; Tadashi Yamamuro ; Hikoto Ohta ; Hiroaki Nakahara ; Shinichi Suzuki
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2015
• 出版时间：October 28, 2015
• 年：2015
• 卷：63
• 期：42
• 页码：9363-9371
• 全文大小：649K
• ISSN：1520-5118

文摘

A method based on liquid chromatography鈥揺lectrospray mass spectrometric analysis of the enzymatically digested amplicons derived from the mitochondrial 16S rRNA gene was established for the discrimination of toxic pufferfish. A MonoBis C18 narrow-bore silica monolith column (Kyoto Monotech) and a Q Exactive mass spectrometer (Thermo Fisher) were employed for separation and detection, respectively. Monoisotopic masses of the oligonucleotides were calculated using Protein Deconvolution 3.0 software (Thermo Fisher). Although a lock mass standard was not used, excellent accuracy (mass error, 0.83 ppm on average) and precision (relative standard deviation, 0.49 ppm on average) were achieved, and a mass accuracy of <2.8 ppm was maintained for at least 180 h without additional calibration. The present method was applied to 29 pufferfish samples, and results were consistent with Sanger sequencing.