Journal
HIGH THROUGHPUT SCREENING: METHODS AND PROTOCOLS, 3RD EDITION
Volume 1439, Issue -, Pages 47-63Publisher
SPRINGER
DOI: 10.1007/978-1-4939-3673-1_3
Keywords
Mass spectrometry; RapidFire (TM); Electrospray (ESI); High-throughput screening; Triple-quadrupole mass spectrometry; Mass charge (m/z); Ionization
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In order to detect a biochemical analyte with a mass spectrometer (MS) it is necessary to ionize the analyte of interest. The analyte can be ionized by a number of different mechanisms, however, one common method is electrospray ionization (ESI). Droplets of analyte are sprayed through a highly charged field, the droplets pick up charge, and this is transferred to the analyte. High levels of salt in the assay buffer will potentially steal charge from the analyte and suppress the MS signal. In order to avoid this suppression of signal, salt is often removed from the sample prior to injection into the MS. Traditional ESI MS relies on liquid chromatography (LC) to remove the salt and reduce matrix effects, however, this is a lengthy process. Here we describe the use of RapidFire (TM) coupled to a triple-quadrupole MS for high-throughput screening. This system uses solid- phase extraction to de-salt samples prior to injection, reducing processing time such that a sample is injected into the MS similar to every 10 s.
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