一种利用RT-HPLC分析乳酸菌产生物胺的方法
首发时间:2009-08-31
摘要:具有脱羧酶活性的乳酸菌可通过氨基酸的脱羧反应产生具有潜在安全风险的生物胺。本文利用脱羧酶培养基初步筛查61株乳酸菌产生物胺情况,再通过RT-HPLC法测定其在发酵液和发酵乳中的生物胺含量。用10%的三氯乙酸提取样品中的生物胺,采用苯甲酰氯衍生处理后,以甲醇/水为流动相,进行梯度洗脱,流速0.8mL/min,紫外检测器波长为254nm。结果显示,组胺和酪胺得到良好的分离,在给定的浓度范围内呈现良好的线性关系(R2>0.995)。在发酵液和发酵乳中添加生物胺混合标准溶液,平均回收率为97.92-101.14%,相对偏差RSD<5%。结果表明,发酵液与发酵乳中生物胺的RT-HPLC法,是一种快捷,稳定,灵敏度高的检测方法,其与脱羧酶培养基法结合可以准确的实现对乳酸菌产生物胺的评价,确保乳酸菌更安全的应用于食品中。
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A RT-HPLC Analysis Method for Biogenic Amine Produced by Lactic Acid Bacteria
Abstract:Lactic acid bacteria with decarboxylase activity, through decarboxylation of amino acids reaction, may produce biogenic amines which have the potential security risks. In this paper, 61 strain lactic acid bacteria were screened by a decarboxylase medium, and then biogenic amines produced by fermented broth and milk was quantitatively detected by the RT-HPLC method. The biogenic amines were extracted by 10% trichloroacetic acid, benzoyl chloride derivation. The mobile phase was a gradient elution program with methanol and water. The flow rate was 0.8ml/min. UV detector was used in the final determination with λ=254nm. The results showed that biogenic amines were separated well. The method was linear for the amines studied at given concentrations (R2>0.995). The average recoveries ranged from 97.92%-110.14 %for all amines samples. The RSDs were less than 5 %. It can be seen that the RT-HPLC method was rapid, steady, high sensitive. The combination with the screening decarboxylase medium could evaluate the safety of Lactic acid bacteria more accurately and ensure their applications in food more securely.
Keywords: Lactic Acid Bacteria biogenic amine decarboxylase RT-HPLC
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