反相高效液相色谱法同时检测海产品中8种生物胺

Simultaneous determination of eight biogenic amines in seafood by reversed-phase high performance liquid chromatography

中文关键词:  反相高效液相色谱法  生物胺  海产品  秋刀鱼  过敏反应

英文关键词:reversed-phase high performance liquid chromatography  biogenic amines  seafood  Cololabis saira  allergic reactions

中文摘要:

      目的 建立一种可同时快速测定海产品中8种生物胺的丹磺酰氯柱前衍生反相高效液相色谱法。方法 样品经5%三氯乙酸提取, 丹磺酰氯柱前衍生后, 采用Agilent zorbax SB-C18色谱柱(4.6 mm×150 mm, 5 μm), 以甲醇和水作为流动相, 进行梯度洗脱。柱温30 ℃, 流速为1.5 mL/min, 采用UV紫外检测器, 检测波长254 nm, 对8种生物胺进行分离分析。结果 结果显示, 色胺、2-苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺在15 min内得到有效分离, 在1~100 mg/L范围内线性系数r2﹥0.999, 8种生物胺测定方法的检出限范围为0.02~0.17 mg/L, 定量限范围为0.067~0.57 mg/L, 且重复性良好, 向秋刀鱼中分别添加生物胺标准品20.0、50.0、100.0 mg/L 3个含量水平, 8种生物胺平均回收率介于79%~117%之间, 相对标准偏差为0.84%~12.57%。采用该方法测定了秋刀鱼等5类海产品中8种生物胺的含量, 结果显示生物胺总含量在23~208 mg/kg之间。结论 该方法简单、快速, 灵敏度和准确度符合要求, 适用于海产品中多种生物胺的同时检测。

英文摘要:

      Objective To establish a method for simultaneous determination of eight biogenic amines in seafood by reversed-phase high performance liquid chromatographic (RP-HPLC) with precolumn derivatization by dansylchloride. Methods Samples were extracted with trichloroacetic acid, and derivatized with dansylchloride. Using Agilent zorbax SB-C18 chromatography column (4.6 mm×150 mm, 5 μm), eight biogenic amines were separated and analyzed by RP-HPLC. The chromatographic conditions of RP-HPLC were as follows: the mobile phase for gradient elution consisted of methanol and water, the flow rate was 1.5 mL/min, the column temperature was 30 ℃, the UV detector was used and the detection wavelength was 254 nm. Results The results showed a good separation of eight biogenic amines (tryptamine, 2-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine) within 15 min. It was linear in the range of 1~100 mg/L (r2﹥0. 999). The detection limits of eight biogenic amines were 0.02~0.17 mg/L, and the limits of quantification were 0.067~0.57 mg/L. The reproducibility of the method was good. The average recoveries of biogenic amines with different mass concentrations were 79%~117%, and the relative standard deviations (RSDs) ranged from 0.84% to 12.57%. The results showed that the total content of biogenic amines were between 23 and 208 mg/kg in five kinds of aquatic products. Conclusion The established method is simple, rapid, accurate, and sensitive, and can be used for simultaneous determination of biogenic amines in aquatic products.