Department of Food Science and Nutrition, School of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, China.
Anal Chem. 2011 May 1;83(9):3297-304. doi: 10.1021/ac1029538. Epub 2011 Apr 4.
We developed an automated microwave digestion labstation (MDL) combined with ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method under the control of positive-negative ion switching as a robust kinetic study tool for rapid and simultaneous quantification of asparagine, glucose, fructose, and acrylamide in Maillard reaction products. Maillard reactions were conducted in a potato model via MDL. The two-step simple pretreatment procedures included the addition of isotope internal standards (15)N(2)-asparagine, (13)C(6)-glucose, and D(3)-acrylamide, followed by appropriate dilution with the mobile phase and filtration. Analytes were separated on a Hypercarb column and monitored by MS/MS. Study of matrix effects indicated Maillard reaction products induce an ionization suppression of both positive and negative precursor ions, but quantitative results are corrected through the use of isotopically labeled internal standards. Using this method, the limit of detection (LOD) and limit of quantification (LOQ) ranges of all analytes were calculated as 0.04-0.6 and 0.1-1.1 μM, respectively. Excellent repeatability (RSD < 9.6%) and acceptable within-laboratory reproducibility (RSD < 9.2%) substantially supported the use of this method for sample analysis. The present kinetic tools, with 10-50 min mimic of Maillard reactions and short instrumental run time (5.5 min per sample), were successfully validated and applied to simultaneous determination of acrylamide and its precursors and intermediates during Maillard reactions and kinetic elucidation. Furthermore, current tools of MDL combined with simple sample treatment procedures and UHPLC-MS/MS analysis reduce sample analysis time and labor in the kinetic study.
我们开发了一种自动化微波消解实验室工作站（MDL），结合正负离子切换的超高效液相色谱-串联质谱（UHPLC-MS/MS）方法，作为一种强大的动力学研究工具，用于快速和同时定量测定美拉德反应产物中天冬酰胺、葡萄糖、果糖和丙烯酰胺。美拉德反应通过 MDL 在马铃薯模型中进行。两步简单的预处理程序包括添加同位素内标（15）N（2）-天冬酰胺、（13）C（6）-葡萄糖和 D（3）-丙烯酰胺，然后用流动相适当稀释并过滤。分析物在 Hypercarb 柱上分离，并通过 MS/MS 监测。基质效应研究表明，美拉德反应产物会抑制正离子和负离子前体离子的离子化，但通过使用同位素标记的内标可以校正定量结果。使用该方法，所有分析物的检测限（LOD）和定量限（LOQ）范围分别计算为 0.04-0.6 和 0.1-1.1 μM。出色的重复性（RSD < 9.6%）和可接受的实验室内重复性（RSD < 9.2%）极大地支持了该方法用于样品分析。这种动力学工具可以模拟 10-50 分钟的美拉德反应，仪器运行时间短（每个样品 5.5 分钟），成功地进行了验证，并应用于美拉德反应过程中丙烯酰胺及其前体和中间体的同时测定和动力学阐明。此外，MDL 与简单的样品处理程序和 UHPLC-MS/MS 分析相结合的当前工具减少了动力学研究中的样品分析时间和劳动力。
