高效液相色谱-荧光检测法测定发酵红肉制品中Neu5Gc

Determination of Neu5Gc in Fermented Red Meat Products by HPLC-FLD

DOI：10.3969/j.issn.1673-1689.2019.05.010

中文关键词: N-羟乙酰神经氨酸 红肉 发酵肉制品 荧光高效液相色谱 食品安全

英文关键词: N-glycolylneuraminic acid,red meat,fermented meat,HPLC-FLD,food safety

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中文摘要:

基于高效液相色谱-荧光检测（HPLC-FLD）技术，建立红肉发酵及制品中N-羟乙酰神经氨酸（N-glycolylneuraminic acid，Neu5Gc）的定量分析方法。成功搭建了以酸水解释放试样中的Neu5Gc，4，5-亚甲二氧基-1，2-邻苯二胺盐（4，5-methylenedioxy-1，2-phenylenediamine dihydrochloride，DMB）为衍生化试剂，并采用安捷伦ZORBAX Eclipse XDB-C18色谱柱等系列条件的HPLC-FLD分析检测方法。该方法经验证：Neu5Gc在25～400 μmol/L范围内与Neu5Gc峰面积的线性关系良好，R2＝0.999 41；样品加标平均回收率在96.4%～99.07%之间；RSD为1.2％，重复性的RSD为1.5％；仪器检出限为0.001 μmol/L，定量限为0.003 μmol/L。检测发酵红肉样品中Neu5Gc的质量分数在9.59～20.08 μg/g之间，不同的发酵红肉制品Neu5Gc的含量之间均存在显著性差异（p＜0.05）。本方法能有效分离Neu5Gc目标峰（分离度R＞1.5）、操作简单、重复性好、灵敏度和精密度高，可广泛用于发酵红肉制品中Neu5Gc的定量测定。

英文摘要:

The establishment of quantitative analysis method of N-glycolylneuraminic acid（Neu5Gc） based on HPLC-FLD technique has become the key to explore the mechanism of Neu5Gc reduce or increase in red meat fermentation and fermented meat products. We released the Neu5Gc in the sample by acid hydrolysis and used 4，5-Methylenedioxy-1，2-phenylenediamine dihydrochloride（DMB） as a derivatization agent. The chromatographic separation was accomplished using an Agilent ZORBAX Eclipse XDB-C18 column，etc. The results showed that the proposed method showed a good linearity between the ratio of Neu5Gc peak area and the mass concentration of Neu5Gc in the range of 25～400 μmol/L，R2＝0.999 41；The recovery of spiked samples was between 96.4%～ 99.07%；RSD was 1.2%；the instrumental had a limit of detection（LOD） of 0.001 μmol/L，and the limit of quantitation（LOQ） was 0.003 μmol/L. The Neu5Gc content of fermented red meat samples was detected to be in the range of 9.59～20.08 μg/g，with significant differences existing among these samples（p＜0.05）. The method can effectively separate the target peaks of Neu5Gc（resolution p＞1.5） with simple operation，good repeatability，high sensitivity and precision，which can be widely used for the determination of Neu5Gc in fermented red meat products.

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