Shanghai Mass Spectrometry Center, Shanghai Institute of Organic Chemistry, Chinese Academy of Science, Shanghai 200032, China.
Anal Chem. 2010 Mar 1;82(5):2113-8. doi: 10.1021/ac9019945.
A selective, fast, and effective enzyme assay based on matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS) for quantifying beta-lactamase, an illegal additive in milk products, has been reported. The strengths of the mass spectrometric assay are its response to all substrate and products, simple and direct detection of the conversion of substrate, and facile determination of enzyme activity. Also, MALDI MS is tolerant to many buffer salts and reagents without the requirement of complicated sample pretreatment procedures. In this study, the approach was used to detect the presence of beta-lactamases (BLA) in milk samples. The amount of BLA that could be determined in a milk sample is 6 x 10(-3) U x mL(-1) by this approach. To test the strategy, it has been applied to the fortified milk (adding a BLA product known as an antimicrobial destroyer). It is then tested whether the pasteurization procedure of the milk process affects the activity of BLA in milk samples. This study offers a perspective into the utility of MALDI-FTMS as an alternative detection tool for BLA screening in milk.
已报道了一种基于基质辅助激光解吸/电离傅里叶变换质谱(MALDI-FTMS)的选择性、快速且有效的酶分析法,用于定量奶制品中的非法添加物——β-内酰胺酶。该质谱分析法的优势在于其对所有底物和产物均有响应,可直接简单地检测底物的转化,并能方便地测定酶活性。此外,MALDI MS 可耐受多种缓冲盐和试剂,无需进行复杂的样品预处理。在本研究中,该方法用于检测牛奶样品中β-内酰胺酶(BLA)的存在。通过该方法可在牛奶样品中检测到 6 x 10(-3) U x mL(-1)的 BLA。为了测试该策略,已将其应用于强化牛奶(添加一种称为抗菌破坏剂的 BLA 产品)中。然后测试牛奶加工过程中的巴氏杀菌程序是否会影响牛奶样品中 BLA 的活性。本研究为 MALDI-FTMS 作为替代检测工具用于牛奶中 BLA 筛查提供了一个视角。