免疫亲和柱净化-高效液相色谱-三重四极杆质谱法测定双壳类水产中软骨藻酸

Determination of domoic acid in bivalve aquatic products by immunoaffinity column purification-high performance liquid chromatography-triple quadrupole mass spectrometry

中文关键词:  双壳类  贝类毒素  软骨藻酸  免疫亲和柱  高效液相色谱-三重四极杆质谱法  基质效应

英文关键词:bivalves  shellfish toxin  domoic acid  immunoaffinity column  high performance liquid chromatography-triple quadrupole mass spectrometry  matrix effect

中文摘要:

      目的 建立免疫亲和柱净化-高效液相色谱-三重四极杆质谱法(liquid chromatography coupled with triple quadrupole mass spectrometry, HPLC-MS/MS)检测双壳类水产中软骨藻酸的方法。方法 样品用80%甲醇水提取, 上清液加入磷酸缓冲液后经免疫亲和柱净化, 将洗脱液氮吹至干定容后上机测定。待测物经InfinityLab Poroshell 120 EC-C18色谱柱分离, 以10 mmol/L甲酸铵-0.1%甲酸-甲醇为流动相进行梯度洗脱。多重态反应监测(multiple reaction monitoring, MRM)模式检测。结果 软骨藻酸在20.0~200 ng/mL范围内线性关系良好, 相关系数为0.9999, 检出限为0.002 μg/g, 回收率为75.2%~85.7%, 相对标准偏差为0.8%~3.4%。结论 本方法特异性强、提取效果好、无基质抑制效应, 适用于双壳类水产中软骨藻酸的痕量检测。

英文摘要:

      Objective To establish a method for determination of domoic acid in bivalve aquatic products by immunoaffinity column purification-high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS). Methods The sample was extracted with 80% methanol water, and the supernatant was added to phosphate buffer and purified by an immunoaffinity column. The eluent was blown to dry and constant volume with nitrogen and then measured on the machine. The analyte was separated with an InfinityLab Poroshell 120 EC-C18 chromatographic column using 10 mmol/L ammonium formate-0.1% formic acid-methanol as the mobile phase for gradient elution, detected with multiple reaction monitoring (MRM) mode. Results Domoic acid had a good linear relationship in the range of 20.0?200 ng/mL, the correlation coefficient was 0.9999, the limit of detection was 0.002 μg/g, the recovery rates were 75.2%?85.7%, and the relative standard deviations were 0.8%?3.4%. Conclusion The method has strong specificity, good extraction effect and no matrix inhibition effect. It is suitable for trace determination of domoic acid in bivalve aquatic products.