SPÁČIL, Zdeněk, S ELLIOTT, SL REEBER, MH GELB, CR SCOTT and F TURECEK. 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. Analytical Chemistry. WASHINGTON: AMER CHEMICAL SOC, vol. 83, No 12, p. 4822-4828. ISSN 0003-2700. doi:10.1021/ac200417u. 2011.
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Original name 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
Authors SPÁČIL, Zdeněk, S ELLIOTT, SL REEBER, MH GELB, CR SCOTT and F TURECEK.
Edition Analytical Chemistry, WASHINGTON, AMER CHEMICAL SOC, 2011, 0003-2700.
Other information
Original language English
Type of outcome Article in a journal
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 5.856
Doi http://dx.doi.org/10.1021/ac200417u
UT WoS 000291499800032
Changed by Changed by: PharmDr. Zdeněk Spáčil, Ph.D., učo 238088. Changed: 29/9/2017 13:52.
Abstract
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 workup. 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-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.
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