柱前衍生HPLC同时测定鱼中多种生物胺及其变化规律

Biogenic Amines in Fish Determined by HPLC with Pre-column Dansylation

目的：采用高效液相色谱法同时测定鱼中7种生物胺（即色胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺）的含量,研究在不同贮藏条件下鱼体生物胺的变化规律。方法：样品用5%三氯乙酸溶液提取后,用丹磺酰氯进行柱前衍生,经C18色谱柱（4.6mmI.D.×150mm）分离,用水和乙腈进行梯度洗脱,于波长254nm处检测。考察大黄鱼和黄鲷在-20、4和25℃贮藏不同时间时生物胺的变化情况。结果：20min内7种生物胺得到良好的分离,在0.1~20.0mg/kg范围具有良好的线性关系（r〉0.999）。生物胺的平均回收率73%~101%,RSD5.9%~20.1%。所测的7种新鲜海鱼中,除金线鱼具有较高的生物胺含量（170.8mg/kg）外,其它鱼的生物胺含量很低（3.95~23.63mg/kg）。在-20℃条件下大黄鱼和黄鲷中生物胺的总量无显著变化（p〈0.05）,而在4℃和25℃时,鱼体中的生物胺总量随贮藏时间的延长而增加。其中腐胺、尸胺、组胺和酪胺含量变化敏锐,随着贮藏时间的延长和温度的上升,其含量显著增加（p〈0.05）。结论：低温贮藏可有效控制鱼体中生物胺的形成。腐胺、尸胺、组胺和酪胺可作为判断鱼体新鲜度的质量指标。

Objective： Seven biogenic amines （including tryptamine, putrescine, cadaverine, hisamine, tyramine, spermidine and spermine） in fish were simultaneously determined by high -performance liquid chromatograph （HPLC）, and the changes of biogenic amines of fish stored at different conditions were investigated. Method： After being extracted with 5% TCA from the aquatic products, the biogenic amines were derived by dansyl chloride before the separation with a C18 column （4.6 mm I.D. ×150 mm）. The biogenic amines were eluted using a mixture of water and acetonitrile at a gradient elution program, and were detected at 254 nm. The changes of biogenic amines in the two species of Pseudosci- aena crocea and Dentex tumifrons separately stored at -20 ℃, 4 ℃ and 25 ℃ for different time were measured. Results： Seven biogenic amines could well be separated within 20 min. A good linear response of biogenic amines ranged from 0.1 to 20 mg/L with γ 0.999. The average recoveries tested by adding standards ranged from 73% to 101% for all amines with RSD from 5.9% to 20.1%. Lower levels of biogenic amines （ranged from 3.95 to 23.63 mg/kg） were found in the seven species of measured fresh sea-fish except for Sinocyclocheilus grahami with the value of 170.8 mg/kg. Results also showed that the amounts of total biogenic amines in the two species of Pseudosciaena crocea and Dentex tumifrons stored at -20 ℃ did not significantly change（p0.05）, while, their values increased when the fish were stored at both 4 ℃ and 25 ℃. In addition, the contents of four biogenic amines including putrescine, cadaverine, hisamine and tyramine significantly increased with the increase of storage time and temperature. Conclusion： The formation of biogenic amines can be efficiently controlled by lower temperature. Putrescine, cadaverine, hisamine and tyramine might be used as the potential freshness indexes for fish.