超高效液相色谱-柱前衍生法同时测定水产品中的8种生物胺

食品科学 ›› 2012, Vol. 33 ›› Issue (18): 181-185.

超高效液相色谱-柱前衍生法同时测定水产品中的8种生物胺

张阳1，吴光红2，刘文斌1,*，孟勇2，黄鸿兵2

1.南京农业大学动物科技学院 2.江苏省淡水水产研究所水产质量检测中心

收稿日期:2011-07-19 修回日期:2012-07-17 出版日期:2012-09-25 发布日期:2012-09-07
通讯作者: 刘文斌 E-mail:wowzhangyang0512@163.com
基金资助:
2010年江苏省水产三项工程水产品中生物胺和杂环胺检测技术研究

Simultaneous Determination of 8 Biogenic Amine Residues in Aquatic Products by Ultra-high Performance Liquid Chromatography

Received:2011-07-19 Revised:2012-07-17 Online:2012-09-25 Published:2012-09-07

摘要/Abstract

摘要： 应用超高效液相色谱检测平台，建立同时测定水产品中8种生物胺的高灵敏度快速检测。水产品肉组织经高氯酸溶液冰冻离心提取后，正己烷除脂，丹磺酰氯溶液衍生。氨水结束反应，乙腈定容，进样量5μL，梯度洗脱，紫外检测波长245nm。结果显示：8种生物胺分别在0.5～20μg/mL范围内线性良好，相关系数r≥0.999；8种生物胺加标回收率为71.99%～97.65%，批内精密度为0.15%～9.89%，批间精密度为1.55%～10.7%(n＝6)；8种生物胺的检出限(RSN＝3)：色胺、苯乙胺15.0μg/kg，尸胺、腐胺、组胺6.0μg/kg，酪胺、精胺、亚精胺20.0μg/kg。该方法快速简便，灵敏度高、重复性和准确性好，13min完成分析，适合水产品中生物胺含量的大批量快速的检测分析，检测效率成倍提高。

关键词: 生物胺, 水产品, 超高效液相色谱

Abstract: A sensitive and rapid method for the simultaneous determination of 8 biogenic amine residues in aquatic products was established using an ultra-high performance liquid chromatography (UPLC) system. Aquatic product samples were extracted with perchloric acid solution, centrifugated, defatted with n-hexane and derivatized with dansyl chloride. The derivative was allowed to react with ammonia water and made up to the desired volume with acetonitrile. A 5 ?L sample was injected and separated in a gradient elution mode. The detection wavelength was set as 245 nm. The results showed that 8 biogenic amines displayed a linear relationship in the range of 0.5–20 μg/mL with correlation coefficients not lower than 0.999. The spike recovery rates of 8 biogenic amines were in the range of 71.99%–97.65% with intra-batch precision RSD of 0.15%–8.89% and inter-batch precision RSD of 1.55%–10.7% (n = 6). The limit of detection was 15.0 μg/kg for Try and Phe, 6.0 μg/kg for Put, Cad and His, and 20.0 μg/kg for Try, Spd and Spm. This method was rapid, simple, sensitive, repeatable and accurate and the analysis process could be completed in 13 min. Thus, it is suitable for the rapid and large-scale determination of biogenic amine residues in aquatic products.

Key words: biogenic amines (BAs), aquatic products, ultra-high performance liquid chromatography (UPLC)

中图分类号:

张阳1，吴光红2，刘文斌1,*，孟勇2，黄鸿兵2. 超高效液相色谱-柱前衍生法同时测定水产品中的8种生物胺[J]. 食品科学, 2012, 33(18): 181-185.

参考文献

[1] Esti M,Volpe G,Massignan L,et al.Determination of amines in fresh and modified atmosphere packaged fruits using electrochemical biosensors[J].Journal of Agricultural and Food Chemistry,1998,46:4233–4237
[2] RawlesDD, Flick GJ , Martin RE, et al.Biogenic amines in fish and shellfish[J]. Advances in food and nutrition research,Academic Press,1996,39:329–365
[3] Yoshinaga D H，Frank H A,et al.Histamine producting bacteria in decomposed skipjacktuna (Katsuwonus pelamis) [J].Applied Environmental Microbialogy, 1982,44(3):447–452
[4] Dingemanse J,Wood N, Guentert T, et al.Clinical pharmacology of moclobemide duringchronic administration of high doses to healthy subjects[J].Psychopharmacology, 1998, 140:164–172.
[5] Mietz J L,Karmas E. Chemical quality index of cannel tuna as determined by HPLC[J].Journal of food science, 1997,42(1):155–158
[6] Mietz J L,Karmas E. Polyamine and histamine content of rockfish,salmon,lobster and shrimp as an indictor of decomposition[J].Analytical Technology Biomedical life science, 1978,61(1):139
[7] Lange J，Wittmann C．Enzyme sengor array for the determination of biogenic amines in food samples[J]．Anal Bioanal Chem, 2002.372:276–283
[8] Lin W C, Lin C E,Lin E C. Capillllary zone electrophoretic separation of biogenic amine:influence of orgnic modifier[J].Journal of Chromatography A , 1996,55(1):142–146
[9] Slemr J，Beyermann K. Determination of biogenic am ines in meat by combined ion-exchange and capillary gas chromatography[J]. Chromatogr A, 1984,283:241–500
[10] Fernandes J O,Ferreira M A. Combined ion-pair extraction and gas chromatography-mass spectrometry for the simultaneous determination of diamines,polyamines and aromatic amines in Port wine and grapejuice[J]. Chromatogr A, 2000,886:183–195
[11] HwangB S，WangJT，ChongYM．A rapid gas chromatographic method for the determ ination of histamine in fish and fish products[J]. Food Chem, 2003,82(2) :329–334
[12] Moret S,Bortolomeazzi R,Lercker G.Improvement ofextraction procedure for biogenic amines in food and their highperformance liquid chromatographic determination[J]. Journal of Chromatography, 1992,591:175–180
[13] Romero R,Gazquez D, Bag M G,et al.Optilnization of Chromatographic parameters for the determination of biogenic amines in wine by reversed-phase highperformance liquid chromatographic determination[J]. Journal of Chromatography A, 2000,871:75–83
[14] Soufieros E H, Bouloumpasi E, Zotou A ,et al .Determination of biogenic amines in Greek wines by HPLC[J]. Food Chemistry, 2007,101:704–716
[15] 食品中生物胺含量的测定.GB/T 5009.208-2008[S].北京：中国标准出版社，2008
[16] 徐振，孟勇，徐辛莲，吴光红.反相高效液相色谱法测定冷却猪肉中生物胺[J].江西农业学报，2008，20（8）：82–84