A validated assay to quantitate serotonin in lamb plasma using ultrahigh-performance liquid chromatography-tandem mass spectrometry: applications with LC/MS3

An ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC/MS/MS) method was developed and validated for the quantification of serotonin (5-HT) in lamb plasma using [ 2 d 4 ]-serotonin ([ 2 d 4 ]-5-HT) as an internal standard. Charcoal-stripped human plasma was used as the blank...

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Published inAnalytical and bioanalytical chemistry Vol. 406; no. 20; pp. 5055 - 5059
Main Authors Szeitz, András, Nguyen, Tuan-Anh T., Riggs, K. Wayne, Rurak, Dan
Format Journal Article
LanguageEnglish
Published Berlin/Heidelberg Springer Berlin Heidelberg 01.08.2014
Subjects
Analytical Chemistry
Animals
Biochemistry
Biological Assay
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
Chromatography, High Pressure Liquid - methods
Food Science
Humans
Laboratory Medicine
Monitoring/Environmental Analysis
Plasma - chemistry
Serotonin - analysis
Sheep
Tandem Mass Spectrometry - methods
LC/MS
Serotonin
Method validation
LC/MS/MS
Lamb plasma
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Summary:An ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC/MS/MS) method was developed and validated for the quantification of serotonin (5-HT) in lamb plasma using [ 2 d 4 ]-serotonin ([ 2 d 4 ]-5-HT) as an internal standard. Charcoal-stripped human plasma was used as the blank matrix during validation, and 5-HT was quantitated using selected reaction monitoring. The UHPLC/MS/MS system consisted of an Agilent 1290 Infinity ultrahigh-performance liquid chromatograph coupled with an AB SCIEX QTRAP® 5500 hybrid linear ion trap triple quadrupole mass spectrometer. The method was validated for accuracy, precision, linearity, lower limit of quantification (LLOQ), selectivity, and other parameters. The LLOQ was 1.0 ng/mL, requiring 100 μL of sample. The method was applied to monitor the 5-HT levels in lamb plasma after the administration of fluoxetine. Tandem mass spectrometry cubed (MS 3 ) experiments were also performed to investigate the fragmentation pattern of 5-HT and [ 2 d 4 ]-5-HT. A liquid chromatography-MS 3 (LC/MS 3 ) method was developed, and the UHPLC/MS/MS and the LC/MS 3 methods were compared for performance. Figure ᅟ
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ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-014-7913-6