柱前衍生-RP-HPLC方法测定水产品中生物胺  

Determination of Biogenic Amines in Aquatic Products by Pre-Column Dansy Lalition Reversed Phase High Performance Liquid Chromatography with Fluorescence Detector

作　　者：李志军[1] 薛长湖[1] 封锦芳[2] 周蕊[3] 周雪飞[3] 吴永宁[3] 赵云峰[3] 

机构地区：[1]中国海洋大学食品科学与工程学院 [2]首都医科大学公共卫生与家庭医学学院,北京100050 [3]中国疾病预防控制中心营养与食品安全所,北京100050

出　　处：《中国海洋大学学报（自然科学版）》2007年第3期427-431,共5页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(30571572);科技部中国-希腊政府间科技合作项目资助

摘　　要：建立同时测定水产品中多种生物胺的丹磺酰氯柱前衍生反相高效液相色谱方法。用5％TCA提取样品中生物胺，正己烷脱脂，正丁醇／氯仿（1／1，v／v）萃取，以1，7二胺庚烷为内标，丹磺酰氯为衍生剂衍生后，采用CAPCELL PAK C18MG（4．6mm I．D．×150mm，5μm）色谱柱，以甲醇／水为流动相，进行梯度洗脱，流速1．5mL／min。荧光检测器的激发波长（Ex）350胁，发射波长（Em）520胁。结果显示：色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、精胺和亚精胺等8种生物胺在30min内完全分离。在给定的浓度范围内，待测生物胺峰面积与内标峰面积之比同其相应浓度呈良好的线性相关（r〉0．99）。仪器重复性良好（RSD〈1．34％），方法重复性也在可接收范围内（RSD〈10％）。除色胺（57．7％）外，其它7种生物胺的样品平均回收率均在94.2％～110．3％之间。结果表明生物胺的HPLC检测方法，分离度好，灵敏度高，可快捷、准确地对水产品中生物胺进行定性和定量检测。A pre-column derivatization reversed-phase high-performance liquid chromatography method was developed for the determination of biogenic amines （tryptamine, phenylethyamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine） in aquatic products with fluorescecse detector. 5% TCA was used to extract amines in aquatic products and chloroform/butanol was used to clean up the extraction, which was degreased by n-hexane prior to derivatization with dansyl chloride. The amines were separated using gradient elution. Mobile phase A was methanol and mobile phase B was water with a flow rate 1.5 ml/min, The column used was a CAPCELL PAK C18 MG （4.6 mm I. D. × 150 mm, 5μm） and gradient elution at contant column tempereture with 30 ℃ under fluorescence detector with Ex 350 nm and Em 520 nm. 1, 7-Diamino- heptane was used as the internal standard. The method was linear for the amines studied at concentrations ranging from 0.05 to 25 mg/L, except for spermidine and spermine, whose concentrations ranged from 0.05 to 15 mg/L. The average recoveries were between 94.2 96 - 110.3 % for all amines except for tryptamine （57. 7% ）, RSD below 10 %. With this method the samples were determined with favorable results. The method can be applied to the determination of amines in aquatic products.

关 键 词：高效液相色谱 生物胺 柱前衍生 水产品 

分 类 号：TS201.2[轻工技术与工程—食品科学] TS254.7[轻工技术与工程—食品科学与工程]