高效液相色谱-串联质谱法测定烤鱼片中的河豚毒素

目的建立高效液相色谱-串联质谱(HPLC-MS/MS)测定烤鱼片中河豚毒素(TTX)的检测方法。方法样品经1%乙酸超声振荡提取15min,提取液加入4倍量的甲醇沉淀水溶性大分子化合物,离心后将上清液挥干,残留物加1mL甲醇复溶,高速离心后取上清液进行HPLC-MS/MS分析检测。色谱分离于HILIC柱上进行,梯度洗脱。采用电喷雾离子源正离子多反应监测(MRM)模式进行定性和定量检测。结果 TTX在5~1 000ng/mL浓度范围内线性关系良好,相关系数为0.999 3,检出限为16μg/kg,回收率为84.6%~98.1%,相对标准偏差(RSD)3.9%~13.7%。结论该方法选择性强、灵敏度高、处理方法简单,适用用于烤鱼片中河豚毒素的测定。

Determination of tetrodotoxin in roasted fish fillets by HPLC-MS/MS

Objective To establish an analytical method for qualitative and quantitative analysis of tetrodotoxin （TTX） in roasted fish fillets using HPLC-MS/MS. Methods Specimens were treated by vortexing and ultrasonic-assist extraction in 1 0/40 acetic acid. The extract was added into 4 volumes of methanol to precipitate soluble macromolecule compounds. After centrifugation, supernatant was evaporated and the residue was redissolved in 1 ml of methanol. After high speed centrifugation the supernatant was subjected by HPLC-MS/MS analysis, Separation was performed on a hydrophilic interaction liquid chromatography （HILIC） column via gradient e- lution. Eieetrospray ionization （ESI） source in positive ion mode and multi-reaction monitoring （MRM） was used for qualitative and quantitative analysis. Results TTX showed good lineage in the range of 5- 1 000 ng/ml , correlation coefficient was 0. 999 3. Limit of detection （LOD, S/N= 3） was 16 μg/kg, recovery efficiency for TTX in roast fish fillet was 84.6 %- 98.1 % with relative standard deviation of 3.9 %- 13.7 %. Conclusion The method established in this study is specific, sensitive, easy to operate, which is suitable for qualitative and quantificative analysis of TTX in roasted fish fillets.