摘要
建立了反相HPLC同时测定色胺、2-苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺8种生物胺的方法,并对腌干鱼制品中生物胺的提取方法进行了优化。1,7-二氨基庚烷为内标,经丹磺酰氯衍生,C18柱(250 mm×4.6 mm,5μm)分离后,使用荧光检测器检测,荧光激发波长为350 nm,发射波长为520 nm;乙腈-乙酸铵-超纯水为流动相进行梯度洗脱,流速为1 mL/min;采用NaOH及饱和NaHCO3调节衍生体系pH,三氯乙酸(TCA)、高氯酸(HClO4)和盐酸(HCl)作为提取介质对腌干鱼制品中生物胺进行提取。结果显示,8种生物胺在30 min内完全分离,并呈良好的线性相关性(R2≥0.998 66);精胺的线性范围为0.5~50.0μg/mL,其余7种生物胺为0.5~100.0μg/mL;最低检测限为0.03~0.10μg/mL;仪器精密度较好(RSD≤1.87%);最佳衍生体系pH为11.0;5%TCA提取生物胺效果最为理想;腌干鱼样品中8种生物胺得到有效检测结果,生物胺总量范围为(91.75±3.11)^(986.43±5.42)mg/kg。该方法适合腌干鱼制品等腌制水产品中8种常见生物胺的快速有效检测。
A reversed -phase high performance liquid chromatographic （HPLC） method was established to determine tryptamine, 2 - phenylethylamine, putrescine, cadaverive, histamine, tyramine, spermidine and spermine in salted - dried fish and the extraction method of biogenic amines was optimized. Biogenie amines in salted - dried fish were derived with dansyl chloride. 1,7 -Diaminoheptane was used as an internal standard（IS）. 8 biogenic amines were separated by C18 column（250 mm ×4.6 mm, 5 μm）, and then detected by fluorescence detector with excitation wavelength of 350 nm and emission wavelength of 520 nm. Gradient elute using a mobile phase composed of acetonitrile, ammonium acetate and ultrapure water was at 1 mL/min flow rate. Sodium hydroxide and saturated sodium bicarbonate were used to adjust pH of derivative system and biogenic amines were extracted with trichloroacetic acid （TCA） or perchloric acid （ HClO4 ） or hydrochloric acid （HCl）. The results indicated that 8 biogenic amines completely separated within 30 min, and the correlation coefficient （ R2 ） was greater than 0. 998 66 ; linear range of spermine was among 0.5 -50.0 μg/mL; linear range of the other biogenic amines were among 0.5 - 100.0 μg/mL. The limits of detection were in the range of 0.03 -0.10 μg/mL. The relative standard deviation （RSD） of precision was lower than 1.87%. The pH 11.0 was most suitable for derivative system. The using of 5% TCA for extracting biogenic amines in salted -dried fish was a more suitable extraction than the others. Biogenic amines in seven salted - dried fish were determined effectively and the values of total biogenic amines were in the range of （91.75 ±3.11 ） （986.43 ± 5.42） mg/kg. So this optimized method is a rapid, efficient detection way for 8 biogenic amines in salted -dried aquatic product.
出处
《中国渔业质量与标准》
2014年第4期41-48,共8页
Chinese Fishery Quality and Standards
基金
国家自然科学基金项目(31371800)
中国水产科学研究院基本科研业务费(2014C05XK01)
广东省海洋渔业科技推广专项(A201301C01
A201201I04)
关键词
腌干鱼制品
生物胺
提取
反相高效液相色谱法
salted - dried fish
biogenic amines
extraction
reversed phase high performance liquid chromatography
