色谱

色谱

• 技术与应用 • 上一篇    

柱前衍生-反相高效液相色谱法测定不同方法煮制的猪肉及其汤汁中的游离氨基酸

顾伟钢1, 张进杰1, 辛梅2 姚燕佳1, 纪蓉1, 吕兵兵1, 陈健初1*   

  1. 1.　浙江大学生物系统工程与食品科学学院, 浙江 杭州 310058；　2.　青岛市渔业技术推广站,　山东　青岛　266071
  • 收稿日期:2011-04-25 修回日期:2011-06-01 出版日期:2011-10-28 发布日期:2011-10-13
  • 通讯作者: 陈健初,博士,副教授,主要从事食品加工和分析检测方面的研究.
  • 基金资助:
    黑龙江省自然科学基金项目(No. B200909)和哈尔滨师范大学科技发展预研项目(No. 08XYG-15

Rapid determination of trace iodate using monolithic column ion-pair chromatography coupled with direct conductivity detection

GU Weigang1, ZHANG Jinjie1, XIN Mei2, YAO Yanjia1, JI Rong1, L Bingbing1, CHEN Jianchu1*   

  1. College of Biosystem Engineering and Food Science, Zhejiang University, Hangzhou 310058, China; 2. Qingdao Centre of Popularization of Fisheries Technology, Qingdao 266071, China
  • Received:2011-04-25 Revised:2011-06-01 Online:2011-10-28 Published:2011-10-13

PDF

476

摘要： 建立了反相高效液相色谱(RP-HPLC)分离检测用不同方法煮制的猪肉及其汤汁中17种游离氨基酸的方法。样品经6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC)柱前衍生后,采用Nova-PakTMC18色谱柱分离,以AccQ\5Tag Eluent A稀释液、乙腈和超纯水为流动相,梯度洗脱,检测波长为248 nm,在47 min内实现了17种氨基酸衍生物的基线分离。各氨基酸在1～100 μmol/L(胱氨酸在0.5～50 μmol/L)范围内呈现出良好的线性关系,相应的线性相关系数(r2)均大于0.99; 17种氨基酸的检出限（以信噪比为3计）为0.29～0.96 μmol/L;在汤汁中的加标回收率为86.5%～101.0%。该方法前处理过程简单,分离效果好,是检测肉及其汤汁中氨基酸的有效手段,可应用于肉制品的质量评定和工艺优化。

关键词: 反相高效液相色谱, 汤汁, 游离氨基酸, 猪肉, 柱前衍生

Abstract: A method was developed on a monolithic column for the fast determination of trace iodate (IO~3) by ion-pair chromatography with direct conductivity detection. The analytes were separated using a mobile phase of tetrabutylammonium hydroxide (TBA)-phthalic acid-acetonitrile on a reversed-phase silica-based monolithic column. The effects of eluent, flow rate and column temperature on the retention of iodate were investigated. The optimized chromatographic conditions for the determination of the anion were as follows: 0.25 mmol/L TBA-0.18 mmol/L phthalic acid-3% acetonitrile (pH 5.5) as mobile phase, a flow rate of 4.0 mL/min and a column temperature of 30 ℃. Under the optimal conditions, retention time of iodate was less than 0.5 min and the baseline separation of iodate was achieved without any interference by other anions (Cl~, NO~3, SO2~4, I~). The detection limit (S/N=3) was 0.36 mg/L for IO~3. Relative standard deviation (RSD, n=5) of chromatographic peak area and retention time were 0.35% and 0.28%, respectively. The proposed method was applied to the determination of trace iodate in iodized medicine. The spiked recovery of iodate was 96.4%. The method is rapid, simple, accurate, reliable, and practical.

Key words: broth, free amino acids, high performance liquid chromatography (HPLC), pork, pre-column derivatization