Comparative triplex tandem mass spectrometry assays of lysosomal enzyme activities in dried blood spots using fast liquid chromatography: application to newborn screening of Pompe, Fabry, and Hurler diseases.
Comparative triplex tandem mass spectrometry assays of lysosomal enzyme activities in dried blood spots using fast liquid chromatography: application to newborn screening of Pompe, Fabry, and Hurler diseases.
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DOI：
10.1021/ac200417u
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发表时间：
2011-06-15
影响因子：
7.4
通讯作者：
Turecek, Frantisek
中科院分区：
文献类型：
--
作者：
Spacil, Zdenek;Elliott, Susan;Reeber, Steven L.;Gelb, Michael H.;Scott, C. Ronald;Turecek, Frantisek
我们报告了一项基于三重串联质谱 (MS/MS) 的比较研究，用于检测干血斑中的溶酶体酶，以早期检测新生儿的庞贝病、法布里病和胡勒病。已评估了四种在样品处理和所用设备方面不同的方法。一种新开发的方法使用乙腈进行测定淬灭来沉淀血液蛋白，然后在能够在两个平行色谱柱上进行多次连续样品注射的 LC-电喷雾/MS/MS 系统上进行分析。该方法每次对酶产品和内标进行三重分析需要 1.5 分钟，这与之前报道的流动注射方法的通量相匹配。 LC 分离减少了基质效应，使样品处理更加容易。基于液相色谱的新方法显示出优于目前使用的基于液液萃取到乙酸乙酯中并流动注射到质谱仪中的方法的品质因数。我们研究进行全面比较的其他方法包括液-液萃取到乙酸乙酯中，然后进行 LC-ESI/MSMS，以及乙腈淬灭，然后进行直接流动注射。发现使用乙腈淬火的两种方法都很稳健，并且提供了良好的质量数据，同时比提取方法需要更少的液体转移步骤以及更少的一次性材料和劳动力。讨论了新方法的各自优点，以提出一种经过评估的新生儿筛查实验室高通量分析替代方法。
We report a comparative study of triplex tandem mass spectrometry (MS/MS) based assays of lysosomal enzymes in dried blood spots for the early detection of Pompe, Fabry, and Hurler diseases in newborns. Four methods have been evaluated that differed in sample handling and the equipment used. A newly developed method uses assay quenching with acetonitrile to precipitate blood proteins followed by analysis on an LC-electrospray/MS/MS system capable of multiple consecutive sample injections on two parallel chromatographic columns. This method requires 1.5 min per a triplex analysis of enzyme products and internal standards, which matches the throughput of the previously reported flow injection method. LC separation reduces matrix effects and allows for more facile sample work-up. The new LC-based method showed figures of merit that were superior to those of the currently used method based on liquid-liquid extraction into ethyl acetate and flow injection into the mass spectrometer. The other methods we investigated for comprehensive comparison involved liquid-liquid extraction into ethyl acetate followed by LC-ESI/MSMS, and acetonitrile quenching followed by direct flow injection. Both methods using acetonitrile quenching were found to be robust and provide good quality data while requiring fewer liquid transfer steps and less disposable material and labor than did the extraction methods. The individual merits of the new methods are discussed to present an evaluated alternative approach to high throughput analysis in newborn screening laboratories.
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影响因子：
2
作者：
Jemal, M;Schuster, A;Whigan, DB
通讯作者：
Whigan, DB
影响因子：
9.3
作者：
Duffey, Trisha A.;Bellamy, Garland;Scott, C. Ronald
通讯作者：
Scott, C. Ronald
影响因子：
9.3
作者：
Li, YJ;Scott, CR;Gelb, MH
通讯作者：
Gelb, MH
影响因子：
9.3
作者：
De Jesus, Victor R.;Zhang, X. Kate;Hannon, W. Harry
通讯作者：
Hannon, W. Harry
影响因子：
6.1
作者：
Spacil, Zdenek;Novakova, Lucie;Solich, Petr
通讯作者：
Solich, Petr