微波水解-液相色谱串联质谱法测定16种氨基酸的含量

Determination of 16 kinds of amino acids by microwave hydrolysis-liquid chromatography tandem mass spectrometry

中文关键词:  微波水解  液相色谱-串联质谱法  保健品  氨基酸

英文关键词:microwave hydrolysis  liquid chromatography-tandem mass spectrometry  health food  amino acid

中文摘要:

      目的 建立微波水解-液相色谱串联质谱法(liquid chromatography tandem mass spectrometry, LC-MS)测定保健食品中16种氨基酸的含量。方法 取适量样品, 加入6 mol/L盐酸溶液15 mL, 加入适量苯酚, 于150 ℃微波消解4 h, 冷却稀释至80 ℃减压干燥1 h。样品经0.1%甲酸溶解后经Agilent-C18柱分离, 以乙腈- 10 mmol/L乙酸铵溶液为流动相进行梯度洗脱, 以串联质谱的多反应监测模式进行检测, 外标法定量。结果 方法的线性范围0.01~0.05 nmol/μL; 平均回收率为95.6%~100.7%, 相对标准偏差(relative standard deviation, RSD)小于3.0%。结论 该方法分析速度快、灵敏度高、准确和重复性好, 与现有技术对比, 该方法通过微波技术辅助水解, 缩短了样品的前处理时间, 可适用于保健食品中16种氨基酸的快速检测。

英文摘要:

      Objective To establish a method for determination of 16 kinds of amino acids in health food by microwave hydrolysis-liquid chromatography tandem mass spectrometry. Methods Adding 15 mL 6 mol/L hydrochloric acid solution and appropriate amount of phenol to sample, it was hydrolyzed by microwave at 150 ℃ for 4 h, cooled to 80 ℃ and dried for 1 h under reduced pressure. The sample was dissolved in 0.1% formic acid and separated by Agilent-C18 column. The gradient elution was performed with acetonitrile-10 mmol/L ammonium acetate as the mobile phase. The sample was detected by multi-reaction monitoring mode of tandem mass spectrometry and quantified by external standard method. Results The linear range of the method was 0.01-0.05 nmol/μL; the average recovery was 95.6%-100.7%, and the relative standard deviation (RSD) was less than 3.0%. Conclusion This method is rapid, sensitive, accurate and repeatable. Comparing with the current technology, it is assisted by microwave hydrolysis to shorten the sample pretreatment time, which can be used for the simultaneous determination of 16 amino acids in health food.