Horie Masakazu, Ishii Rie, Kobayashi Susumu, Nakazawa Hiroyuki
Saitama Prefectural Institute of Public Health: 639-1, Kamiokubo, Saitama, Saitama 338-0824, Japan.
Shokuhin Eiseigaku Zasshi. 2002 Aug;43(4):234-8. doi: 10.3358/shokueishi.43.234.
A simple and reliable method using liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS) for the analysis of tetrodotoxin in puffer-fish was developed. Tetrodotoxin in puffer-fish was extracted with 0.1% acetic acid by heating in a boiling water bath, and the extracts were cleaned up on a Bond Elut C18 (500 mg) cartridge. The LC separation was performed on a TSK-gel ODS 80Ts column (25 cm x 2 mm i.d.) using 5 mmol/L heptafluoro-n-butyric acid (HFBA)-methanol (99:1) as the mobile phase at a flow rate of 0.2 mL/min. Positive ionization produced a typical [M + H]+ molecular ion of tetrodotoxin (m/z 320.1). The calibration graph for tetrodotoxin was rectilinear from 0.1 to 1 microgram/mL with selected ion monitoring (SIM). The detection limit of tetrodotoxin in puffer-fish was 1 microgram/g (equivalent to ca. 5 MU/g).
开发了一种使用液相色谱/电喷雾电离-质谱法（LC/ESI-MS）分析河豚中毒素的简单可靠方法。河豚中的毒素通过在沸水浴中加热，用0.1%乙酸提取，提取物在Bond Elut C18（500mg）柱上进行净化。液相色谱分离在TSK-gel ODS 80Ts柱（25cm×2mm内径）上进行，使用5mmol/L七氟正丁酸（HFBA）-甲醇（99:1）作为流动相，流速为0.2mL/min。正离子化产生了典型的河豚毒素[M + H]+分子离子（m/z 320.1）。在选择离子监测（SIM）模式下，河豚毒素的校准曲线在0.1至1微克/毫升范围内呈线性。河豚中毒素的检测限为1微克/克（相当于约5MU/克）。
