National Research Council Canada, Ottawa, Ontario, Canada.
Anal Chim Acta. 2012 Sep 26;744:54-9. doi: 10.1016/j.aca.2012.07.011. Epub 2012 Jul 16.
A simple, rapid microwave digestion procedure for protein hydrolysis preceding the determination of amino acids in yeast using gas chromatography-mass spectrometry (GC-MS) is described. Protein hydrolysis was performed in a focused microwave using 4M methanesulfonic acid (MAS). Amino acids were derivatized with methyl chlorofomate (MCF) and extracted into chloroform prior to GC-MS analysis. The microwave parameters, including power, temperature and heating time, were optimized. It was found that temperature and heating time were the most influential factors. A total of 17 amino acids were determined in selenium-enriched yeast with use of standard addition calibration. Limits of detection and quantitation (LODs/LOQs) of the amino acids measured were in the sub-nmol range, suitable for monitoring of amino acids in yeast and other food products.
本文描述了一种使用气相色谱-质谱联用仪(GC-MS)测定酵母中氨基酸前对蛋白质进行水解的简单、快速的微波消解方法。在聚焦微波中使用 4M 甲磺酸(MAS)进行蛋白质水解。在进行 GC-MS 分析之前,用甲基氯仿酸盐(MCF)对氨基酸进行衍生化,并萃取到氯仿中。优化了微波参数,包括功率、温度和加热时间。结果发现,温度和加热时间是最具影响力的因素。使用标准添加校准法测定了富硒酵母中的 17 种氨基酸。所测氨基酸的检测限和定量限(LOD/LOQ)均在亚纳摩尔范围内,适用于酵母和其他食品中氨基酸的监测。