高效液相色谱法测定植物源性食品中γ-氨基丁酸含量

Determination of γ-amino butyric acid content in foods of plant origin by high performance liquid chromatography

中文关键词:  高效液相色谱法  植物源性食品  γ-氨基丁酸

英文关键词:high performance liquid chromatography  foods of plant origin  γ-amino butyric acid  4-dimethylamino-azobenzene 4-sulfonyl chloride

中文摘要:

      目的 建立高效液相色谱法测定植物源性食品中γ-氨基丁酸含量的分析方法。方法 以4-二甲基胺基偶氮苯-4-磺酰氯为柱前衍生试剂, 以提取溶剂、料液比、提取方式和提取时间为考察因素, 设计正交实验并进行方法学验证。通过检测不同植物源性食品、仪器比对测定, 分析γ-氨基丁酸测定结果。结果 确定提取γ-氨基丁酸最优条件为60%乙醇溶液, 按1:25 (g:mL)的料液比, 通过水浴振荡(40 ℃)或超声提取30 min。色谱条件: C18色谱柱(250 mm×4.6 mm, 5 μm), 柱温30 ℃, 流动相为乙腈+乙酸钠溶液(20:80, V:V), 流速1.0 mL/min, 检测波长为436 nm。γ-氨基丁酸在1.092~131.064 μg/mL范围内与峰面积具有良好的线性关系。火龙果(白心)、番茄和麦芽中γ-氨基丁酸精密度的相对标准偏差均小于5.0%, 加标回收率均在95.80%~104.36%之间, 表明方法精密度良好, 添加回收率高。结论 本研究建立的方法重复性好、准确度高, 可以准确定量植物源性食品中γ-氨基丁酸的含量。

英文摘要:

      Objective To establish an analytical method for determining γ-amino butyric acid content in foods of plant origin by high performance liquid chromatography. Method Foods samples were pre-column derivatized with 4-dimethylamino-azobenzene 4-sulfonyl chloride, orthogonal experiments were designed and methodological verification was carried out based on extraction solvent, solid-liquid ratio, extraction method and extraction time. The results of γ-amino butyric acid were analyzed by detecting different plant-derived foods and comparing with instruments. Results The results showed that the optimal conditions were determined as 60% ethanol solution, which was extracted by water bath (40 ℃) or ultrasonic for 30 min at 1:25 (g:mL) solid-liquid ratio. The separation of γ-amino butyric acid was achieved on C18 column (250 mm×4.6 mm, 5 μm) using acetonitrile+sodium acetate solution (20:80, V:V) as the mobile phase at a flow rate of 1.0 mL/min. The absorption wavelength of γ-amino butyric acid was 436 nm and column temperature was set at 30 ℃, respectively. The linear range of γ-amino butyric acid was 1.092–131.064 μg/mL. The relative standard deviation of γ-amino butyric acid was all less than 5.0% in pitaya (white heart), tomato, and malt. The average recovery rate of γ-amino butyric acid was between 95.80%–104.36%, indicating that the method had good precisions and high recoveries. Conclusion The method established in this study has good repeatability and high accuracy, and can accurately quantify the content of γ-amino butyric acid in foods of plant origin.