2000
DOI: 10.1016/s0021-9673(99)01292-3
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High-performance liquid chromatography with on-line coupled UV, mass spectrometric and biochemical detection for identification of acetylcholinesterase inhibitors from natural products

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Cited by 194 publications
(127 citation statements)
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“…The analysis rate of the segmented flow method compares favorably with previously reported flow injection AchE assays [26,27,29]. The speed of flow injection methods was limited by the need to inject individual samples or additional separation steps when assay buffer was not directly compatible with ESI-MS.…”
Section: Segmented Flow Esi-ms Analysis For Rapid Screeningmentioning
confidence: 63%
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“…The analysis rate of the segmented flow method compares favorably with previously reported flow injection AchE assays [26,27,29]. The speed of flow injection methods was limited by the need to inject individual samples or additional separation steps when assay buffer was not directly compatible with ESI-MS.…”
Section: Segmented Flow Esi-ms Analysis For Rapid Screeningmentioning
confidence: 63%
“…It may not always be possible to design assays that allow direct injection into an ESI source. Current flow injection systems can utilize in-line solid-phase extraction at high flow rates so their throughput is minimally impacted by this necessity [26]. Therefore, for the segmented flow approach to be as versatile as the flow injection approach for HTS, it is necessary to develop systems that allow compatible solid-phase extraction or similar sample preparation steps that are high-throughput.…”
Section: Overall Throughputmentioning
confidence: 99%
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“…Such biochemical assays are of more interest to the pharmaceutical industry as many drugs target enzymes as part of their mode of action (van Elswijk and Irth 2003;Ingkaninan et al 2000;Schenk et al 2003;Schobel et al 2001). Within our group in cooperation with the Department of Toxicology of Wageningen University, we have developed an on-line HPLC system for detection of Cytochrome P450 (CYP450) 1A2 inhibitors.…”
Section: High-resolution Screeningmentioning
confidence: 99%
“…The assay for measuring acetylcholinesterase activity was based from the assay described by Ellman et al (1961) and Ingkaninan et al (2000) with modifications. Briefly, 125 μL of 3 mM DTNB, 50 μL of sodium phosphate buffer (pH 8.0), 25 μL of the sample dissolved in DMSO and 25 μL of 0.5 U/mL acetylcholinesterase were added in a 96-well microplate and incubated for 15 min at 25°C.…”
Section: Acetylcholinesterase Enzyme Inhibitory Activitymentioning
confidence: 99%