建立了柱前衍生-高效液相色谱(HPLC)法同时检测啤酒中8种生物胺的方法,并研究了啤酒酿造原料、酿造过程及腐败菌的生物胺形成规律.样品经5％三氯乙酸溶液提取后,经丹磺酰氯衍生,然后用HPLC进行定性和定量分析.该方法对8种生物胺具有良好的分离效果,并具有线性范围广,精密度和重复性高等优点,适用于啤酒中生物胺的检测.运用该方法对啤酒酿造原料(麦芽和酒花)中生物胺进行检测,发现除β-苯乙胺外其他7种生物胺均有检出,其中色胺和腐胺含量最高.麦汁和发酵过程中主要含有色胺和腐胺,主要来源于原料麦芽.将2株啤酒腐败乳酸菌-乳酸短杆菌(Lactobacillus brevis)BS49和类布氏乳杆菌(Lactobacillus parabuchneri)B S201接入到无菌啤酒中,分析其生物胺形成规律.这2株菌能在啤酒中生长繁殖,并且随着菌体生长啤酒中生物胺含量发生显著变化,接BS49号菌的啤酒中酪胺由0.2 mg/L增加到8 mg/L,接BS201号菌的啤酒中组胺由0.1 mg/L增加到6 mg/L,因此组胺和酪胺可作为判断啤酒在贮藏过程中是否污染乳酸菌的重要指标.%An analytical method for simultaneous determination of 8 biogenic amines in beer by high-performance liquid chromatography (HPLC) has been developed, meanwhile the forming regularity of biogenic amines in brewing material, brewing process and beer spoilage bacteria was studied.After being extracted with 5％ TCA and derived with dansyl chloride, biogenic amines in beer were qualitatively and quantitatively analyzed by HPLC.The method of HPLC was conformed with efficient separation, good reproducibility, high sensitivity and wide linear range, which was conducive to analysis of biogenic amines in beer.The established method has been successfully applied to determine biogenic amines in malt and hop.The results showed that the 8 biogenic amines were all detected except β-phenylethyl-amine, and the contents of trypatamine and putrescine were the highest.Wort and fermented wort mainly contain trypatamine and putrescine, which come from beer brewing materials.Forming regularity of biogenic amines in spoilage beer by Lactobicillus brevis BS49 and Lactobacillus parabuchneri BS201 were analyzed.Both BS49 and BS201 can grow and propagate in beer, cause the change of content of biogenic amines in spoilage beer.The content of tyramine was increased from 0.1 mg/L to 8 mg/L in spoilage beer by adding BS49, and the content of histamine was increased from 0.1 mg/L to 6 mg/ L in spoilage beer by adding BS201, so it should be seen as an important index for beer spoilage of Lactic acid bacteria.
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