Zhang Xiu-yao, Cai Xin-xin
Institute of Physical and Chemical Inspection, Wenzhou Center for Disease Control and Prevention, Wenzhou 325001, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2013 Mar;47(3):274-7.
To develop a rapid hilic ultra performance liquid chromatography (UPLC)-mass spectrum (MS)/MS method for determination of tetrodotoxin in seafood.
The sample of muscle and liver of puffer fish and nassarius were extracted with aqueous solution containing 0.2% (V/V) acetic acid (the extract of liver must be purified through HLB cartridge), and then cleanup of extract was accomplished by solid-phase extraction with a graphitized carbon black cartridge. The analysis of tetrodotoxin was carried out on a chromatographic column (Acquity UPLC BEH Amide, 100 mm×2.1 mm×1.7 µm) with gradient elution of 95% (V/V) acetonitrile-H2O both containing 0.1% (V/V) formic acid and 2.0 mmol/L ammonium formate, and detected by positive electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode, and quantified by matrix-match standard solution.
The calibration curves were linear in the range of 30 - 10 000, 50 - 10 000 and 30 - 10 000 µg/kg of tetradotoxin in muscle and liver of puffer fish and in muscle of nassarius, respectively. The correlation coefficients were within 0.9963 - 0.9990. The limits of detection were 10, 20 and 10 µg/kg, and that of quantitation were 30, 50 and 30 µg/kg for muscle and liver of puffer fish and muscle of nassarius, respectively. The average recoveries were 81.5% - 93.1%, 82.3% - 106.0% and 83.5% - 95.2% for tetrodotoxin spiked in muscle and liver of puffer fish and in muscle of nassarius, respectively, with relative standard deviation (RSD) of 2.3% - 11%, 4.3% - 14.0% and 3.5% - 13.0% (n = 6).
The method was simple, accurate and sensitive, and could be successfully applied to the measurement of tetrodotoxin in puffer fish and nassarius.
建立一种快速的亲水作用超高效液相色谱（UPLC）-质谱（MS）/MS法测定海产品中的河豚毒素。
用含0.2%（V/V）乙酸的水溶液提取河豚和织纹螺的肌肉及肝脏样品（肝脏提取物须经HLB柱净化），然后用石墨化炭黑柱进行固相萃取净化提取物。在色谱柱（Acquity UPLC BEH酰胺柱，100 mm×2.1 mm×1.7 µm）上，以含0.1%（V/V）甲酸和2.0 mmol/L甲酸铵的95%（V/V）乙腈-水进行梯度洗脱分析河豚毒素，采用正电喷雾电离串联质谱在多反应监测（MRM）模式下检测，并用基质匹配标准溶液进行定量。
河豚肌肉和肝脏以及织纹螺肌肉中河豚毒素的校准曲线在30 - 10 000、50 - 10 000和30 - 10 000 µg/kg范围内呈线性，相关系数在0.9963 - 0.9990之间。河豚肌肉和肝脏以及织纹螺肌肉的检测限分别为10、20和10 µg/kg，定量限分别为30、50和30 µg/kg。河豚肌肉和肝脏以及织纹螺肌肉中添加河豚毒素的平均回收率分别为81.5% - 93.1%、82.3% - 106.0%和83.5% - 95.2%，相对标准偏差（RSD）为2.3% - 11%、4.3% - 14.0%和3.5% - 13.0%（n = 6）。
该方法简便、准确、灵敏，可成功应用于河豚和织纹螺中河豚毒素的测定。
