食品科学

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PITC柱前衍生-反相高效液相色谱法测定黄酒中游离氨基酸和生物胺

芮鸿飞,张晓瑜,刘兴泉*,吴峰华,王卓然,李凯利   

  1. 浙江农林大学农业与食品科学学院,浙江 杭州 311300
  • 出版日期:2016-04-25 发布日期:2016-04-13

Simultaneous Determination of Free Amino Acids and Biogenic Amines in Chinese Rice Wine by RP-HPLC with Pre-Column Phenylisothiocyanate (PITC) Derivatization

RUI Hongfei, ZHANG Xiaoyu, LIU Xingquan*, WU Fenghua, WANG Zhuoran, LI Kaili   

  1. School of Agriculture and Food Science, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China
  • Online:2016-04-25 Published:2016-04-13

摘要:

建立柱前衍生-反相高效液相色谱,同时测定黄酒中17 种氨基酸及7 种生物胺含量的方法。采用异硫氰酸苯酯柱前衍生试剂,采用Inertsil ODS-SP C18(250 mm×4.6 mm,5 μm)分析,以乙酸钠溶液和乙腈溶液混合液进行梯度洗脱,在254 nm波长处进行氨基酸和生物胺的含量分析。氨基酸和生物胺在2.0~500 mg/L和0.5~125 mg/L范围内具有良好的线性关系,相关系数R2大于0.99,平均回收率在86.44%~99.75%之间,相对标准偏差在0.31%~3.53%之间;检出限为0.02~0.25 mg/L,定量限为0.25~4.50 mg/L。方法线性范围广、准确性高、稳定性好,适用于黄酒中氨基酸及生物胺的检测。

关键词: 黄酒, 氨基酸, 生物胺, 反相高效液相色谱

Abstract:

A reversed-phase high performance liquid chromatographic (RP-HPLC) method has been presented for the
simultaneous determination of 17 free amino acids and 7 biogenic amines in Chinese rice wine. Wine samples were precolumn
derivatized with phenylisothiocyanate (PITC), separated using Inertsil ODS-SP C18 chromatography column (250 mm ×
4.6 mm, 5 μm), eluted with a mobile phase composed of sodium acetate, acetonitrile and ultra-pure water, and detected at
254 nm. The calibration curves of 17 amino acids and 7 biogenic amines all exhibited good linearity over the range from 2.0
to 500 mg/L and 0.5 to 125 mg/L, respectively. Mean recoveries for amino acids and biogenic amines were between 86.44%
and 99.75%, and the relative standard deviations (RSDs) were in the range of 0.31%–3.53%. The limits of detection (LODs)
for 17 free amino acids and 7 biogenic amines were in the range of 0.02–0.25 mg/L, and the limits of quantification (LOQs)
were 0.25–4.50 mg/L. The method is stable, accurate, and suitable for the simultaneous determination of amino acids and
biogenic amines in Chinese rice wine, having a wide linear range.

Key words: Chinese wice wine, amino acids, biogenic amines, reversed-phase high performance liquid chromatography (RP-HPLC)

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