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 D
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- 作者：Zdene虒k Spa虂č ; il ; Susan Elliott ; Steven L. Reeber ; Michael H. Gelb ; C. Ronald Scott ; Frantis虒ek Turec虒ek
- 刊名：Analytical Chemistry
- 出版年：2011
- 出版时间：June 15, 2011
- 年：2011
- 卷：83
- 期：12
- 页码：4822-4828
- 全文大小：834K
- 年卷期：v.83,no.12(June 15, 2011)
- ISSN：1520-6882
文摘
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鈥揺lectrospray/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 workup. The new LC-based method showed figures of merit that were superior to those of the currently used method based on liquid鈥搇iquid extraction into ethyl acetate and flow injection into the mass spectrometer. The other methods we investigated for comprehensive comparison involved liquid鈥搇iquid extraction into ethyl acetate followed by LC鈥揈SI-MS/MS 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.