Measurement of serotonin in platelet depleted plasma by liquid chromatography tandem mass spectrometry
5-Hydroxytryptamine (5-HT) in human platelet depleted plasma (PDP) is a biomarker in functional gastrointestinal disorders (FGID), with levels reflecting acute changes in circulating 5-HT concentration. PDP 5-HT is currently measured by reversed phase high performance liquid chromatography (HPLC) fl...
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| Veröffentlicht in: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2009-07, Vol.877 (22), p.2163-2167 |
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| creator | Monaghan, Phillip J. Brown, Heather A. Houghton, Lesley A. Keevil, Brian G. |
| description | 5-Hydroxytryptamine (5-HT) in human platelet depleted plasma (PDP) is a biomarker in functional gastrointestinal disorders (FGID), with levels reflecting acute changes in circulating 5-HT concentration. PDP 5-HT is currently measured by reversed phase high performance liquid chromatography (HPLC) fluorimetric detection. We have developed a simple and rapid liquid chromatography tandem mass spectrometry (LC–MS/MS) method that is two times more rapid than the current HPLC methodology. Our method employs a simple protein precipitation requiring no further downstream sample preparation. 10μL of extract was injected directly onto a SecurityGuard SCX cation exchange column followed by isocratic elution onto an Onyx Monolithic C18 analytical column and methanolic gradient elution. Eluant was connected directly to a Quattro Premier XE tandem mass spectrometer operating in ES+ mode. We detected multiple reaction monitoring transitions m/z 160>114.9 and m/z 164.1>118.9 for 5-HT and d4-5-HT, respectively. 5-HT and d4-5-HT co-eluted at 2.79min and cycle time between injections was 6min. Mean recovery was 98%, limit of detection 1.5nmol/L, lower limit of quantification 5nmol/L, linearity to 1000nmol/L (r2=0.999), imprecision |
| doi_str_mv | 10.1016/j.jchromb.2009.05.045 |
| format | Article |
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PDP 5-HT is currently measured by reversed phase high performance liquid chromatography (HPLC) fluorimetric detection. We have developed a simple and rapid liquid chromatography tandem mass spectrometry (LC–MS/MS) method that is two times more rapid than the current HPLC methodology. Our method employs a simple protein precipitation requiring no further downstream sample preparation. 10μL of extract was injected directly onto a SecurityGuard SCX cation exchange column followed by isocratic elution onto an Onyx Monolithic C18 analytical column and methanolic gradient elution. Eluant was connected directly to a Quattro Premier XE tandem mass spectrometer operating in ES+ mode. We detected multiple reaction monitoring transitions m/z 160>114.9 and m/z 164.1>118.9 for 5-HT and d4-5-HT, respectively. 5-HT and d4-5-HT co-eluted at 2.79min and cycle time between injections was 6min. Mean recovery was 98%, limit of detection 1.5nmol/L, lower limit of quantification 5nmol/L, linearity to 1000nmol/L (r2=0.999), imprecision <10% and bias <13.4%. 5-HT eluted with no ion suppression. No interference was found with l-tryptophan or 5-hydroxyindoleacetic acid (5-HIAA). This assay was compared to a previously published HPLC method. Passing-Bablok regression analysis showed LC–MS/MS=0.91 (HPLC)–0.83, r2=0.97, n=80. Bland Altman analysis showed general agreement, with a mean bias of 3.3nmol/L. We have developed a simple and robust assay for PDP 5-HT that will increase throughput for clinical trials.</description><identifier>ISSN: 1570-0232</identifier><identifier>EISSN: 1873-376X</identifier><identifier>DOI: 10.1016/j.jchromb.2009.05.045</identifier><identifier>PMID: 19520623</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Analysis ; Analytical, structural and metabolic biochemistry ; Biological and medical sciences ; Blood Platelets - metabolism ; Chromatography, Liquid - methods ; Functional gastrointestinal disorders ; Fundamental and applied biological sciences. Psychology ; Gastrointestinal Diseases - diagnosis ; Gastrointestinal Diseases - metabolism ; General pharmacology ; Humans ; LC–MS/MS ; Medical sciences ; Pharmacology. Drug treatments ; Protein precipitation ; Serotonin ; Serotonin - blood ; Serotonin - chemistry ; Serotonin - metabolism ; Tandem Mass Spectrometry - methods</subject><ispartof>Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2009-07, Vol.877 (22), p.2163-2167</ispartof><rights>2009 Elsevier B.V.</rights><rights>2009 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c453t-7fe053c5db5f2ed839546721373d4a421f10325fb9bcf1b35282d33dbfb7ebc83</citedby><cites>FETCH-LOGICAL-c453t-7fe053c5db5f2ed839546721373d4a421f10325fb9bcf1b35282d33dbfb7ebc83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jchromb.2009.05.045$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,45974</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=21691607$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19520623$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Monaghan, Phillip J.</creatorcontrib><creatorcontrib>Brown, Heather A.</creatorcontrib><creatorcontrib>Houghton, Lesley A.</creatorcontrib><creatorcontrib>Keevil, Brian G.</creatorcontrib><title>Measurement of serotonin in platelet depleted plasma by liquid chromatography tandem mass spectrometry</title><title>Journal of chromatography. B, Analytical technologies in the biomedical and life sciences</title><addtitle>J Chromatogr B Analyt Technol Biomed Life Sci</addtitle><description>5-Hydroxytryptamine (5-HT) in human platelet depleted plasma (PDP) is a biomarker in functional gastrointestinal disorders (FGID), with levels reflecting acute changes in circulating 5-HT concentration. PDP 5-HT is currently measured by reversed phase high performance liquid chromatography (HPLC) fluorimetric detection. We have developed a simple and rapid liquid chromatography tandem mass spectrometry (LC–MS/MS) method that is two times more rapid than the current HPLC methodology. Our method employs a simple protein precipitation requiring no further downstream sample preparation. 10μL of extract was injected directly onto a SecurityGuard SCX cation exchange column followed by isocratic elution onto an Onyx Monolithic C18 analytical column and methanolic gradient elution. Eluant was connected directly to a Quattro Premier XE tandem mass spectrometer operating in ES+ mode. We detected multiple reaction monitoring transitions m/z 160>114.9 and m/z 164.1>118.9 for 5-HT and d4-5-HT, respectively. 5-HT and d4-5-HT co-eluted at 2.79min and cycle time between injections was 6min. Mean recovery was 98%, limit of detection 1.5nmol/L, lower limit of quantification 5nmol/L, linearity to 1000nmol/L (r2=0.999), imprecision <10% and bias <13.4%. 5-HT eluted with no ion suppression. No interference was found with l-tryptophan or 5-hydroxyindoleacetic acid (5-HIAA). This assay was compared to a previously published HPLC method. Passing-Bablok regression analysis showed LC–MS/MS=0.91 (HPLC)–0.83, r2=0.97, n=80. Bland Altman analysis showed general agreement, with a mean bias of 3.3nmol/L. We have developed a simple and robust assay for PDP 5-HT that will increase throughput for clinical trials.</description><subject>Analysis</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Blood Platelets - metabolism</subject><subject>Chromatography, Liquid - methods</subject><subject>Functional gastrointestinal disorders</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gastrointestinal Diseases - diagnosis</subject><subject>Gastrointestinal Diseases - metabolism</subject><subject>General pharmacology</subject><subject>Humans</subject><subject>LC–MS/MS</subject><subject>Medical sciences</subject><subject>Pharmacology. Drug treatments</subject><subject>Protein precipitation</subject><subject>Serotonin</subject><subject>Serotonin - blood</subject><subject>Serotonin - chemistry</subject><subject>Serotonin - metabolism</subject><subject>Tandem Mass Spectrometry - methods</subject><issn>1570-0232</issn><issn>1873-376X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkVuL1TAUhYsozkV_gpIXfWvNpUnaJ5FBHWHEFwXfQi47Tg5t00lS4fx7czxFHwcCO-z97WSxVtO8IrgjmIh3h-5g71OcTUcxHjvMO9zzJ80lGSRrmRQ_n9Y7l7jFlNGL5irnA8ZEYsmeNxdk5BQLyi4b_xV03hLMsBQUPcqQYolLWFA966QLTFCQg7UWcKdOnjUyRzSFhy049FeDLvFX0uv9ERW9OJjRrHNGeQVb6hRKOr5onnk9ZXi51-vmx6eP329u27tvn7_cfLhrbc9ZaaUHzJnlznBPwQ1s5L2QlDDJXK97SjzBjHJvRmM9MYzTgTrGnPFGgrEDu27ent9dU3zYIBc1h2xhmvQCcctKyJ4OkstHQdYLMhImKsjPoE0x5wRerSnMOh0VweqUhDqoPQl1SkJhrmoSde_1_sFmZnD_t3brK_BmB3S2evJJLzbkfxwlYiQCn5S-P3NQffsdIKlsAywWXEjVYOVieETKH3rXq9Q</recordid><startdate>20090715</startdate><enddate>20090715</enddate><creator>Monaghan, Phillip J.</creator><creator>Brown, Heather A.</creator><creator>Houghton, Lesley A.</creator><creator>Keevil, Brian G.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TB</scope><scope>8FD</scope><scope>FR3</scope><scope>7X8</scope></search><sort><creationdate>20090715</creationdate><title>Measurement of serotonin in platelet depleted plasma by liquid chromatography tandem mass spectrometry</title><author>Monaghan, Phillip J. ; Brown, Heather A. ; Houghton, Lesley A. ; Keevil, Brian G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c453t-7fe053c5db5f2ed839546721373d4a421f10325fb9bcf1b35282d33dbfb7ebc83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Analysis</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biological and medical sciences</topic><topic>Blood Platelets - metabolism</topic><topic>Chromatography, Liquid - methods</topic><topic>Functional gastrointestinal disorders</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gastrointestinal Diseases - diagnosis</topic><topic>Gastrointestinal Diseases - metabolism</topic><topic>General pharmacology</topic><topic>Humans</topic><topic>LC–MS/MS</topic><topic>Medical sciences</topic><topic>Pharmacology. Drug treatments</topic><topic>Protein precipitation</topic><topic>Serotonin</topic><topic>Serotonin - blood</topic><topic>Serotonin - chemistry</topic><topic>Serotonin - metabolism</topic><topic>Tandem Mass Spectrometry - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Monaghan, Phillip J.</creatorcontrib><creatorcontrib>Brown, Heather A.</creatorcontrib><creatorcontrib>Houghton, Lesley A.</creatorcontrib><creatorcontrib>Keevil, Brian G.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of chromatography. B, Analytical technologies in the biomedical and life sciences</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Monaghan, Phillip J.</au><au>Brown, Heather A.</au><au>Houghton, Lesley A.</au><au>Keevil, Brian G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Measurement of serotonin in platelet depleted plasma by liquid chromatography tandem mass spectrometry</atitle><jtitle>Journal of chromatography. B, Analytical technologies in the biomedical and life sciences</jtitle><addtitle>J Chromatogr B Analyt Technol Biomed Life Sci</addtitle><date>2009-07-15</date><risdate>2009</risdate><volume>877</volume><issue>22</issue><spage>2163</spage><epage>2167</epage><pages>2163-2167</pages><issn>1570-0232</issn><eissn>1873-376X</eissn><abstract>5-Hydroxytryptamine (5-HT) in human platelet depleted plasma (PDP) is a biomarker in functional gastrointestinal disorders (FGID), with levels reflecting acute changes in circulating 5-HT concentration. PDP 5-HT is currently measured by reversed phase high performance liquid chromatography (HPLC) fluorimetric detection. We have developed a simple and rapid liquid chromatography tandem mass spectrometry (LC–MS/MS) method that is two times more rapid than the current HPLC methodology. Our method employs a simple protein precipitation requiring no further downstream sample preparation. 10μL of extract was injected directly onto a SecurityGuard SCX cation exchange column followed by isocratic elution onto an Onyx Monolithic C18 analytical column and methanolic gradient elution. Eluant was connected directly to a Quattro Premier XE tandem mass spectrometer operating in ES+ mode. We detected multiple reaction monitoring transitions m/z 160>114.9 and m/z 164.1>118.9 for 5-HT and d4-5-HT, respectively. 5-HT and d4-5-HT co-eluted at 2.79min and cycle time between injections was 6min. Mean recovery was 98%, limit of detection 1.5nmol/L, lower limit of quantification 5nmol/L, linearity to 1000nmol/L (r2=0.999), imprecision <10% and bias <13.4%. 5-HT eluted with no ion suppression. No interference was found with l-tryptophan or 5-hydroxyindoleacetic acid (5-HIAA). This assay was compared to a previously published HPLC method. Passing-Bablok regression analysis showed LC–MS/MS=0.91 (HPLC)–0.83, r2=0.97, n=80. Bland Altman analysis showed general agreement, with a mean bias of 3.3nmol/L. We have developed a simple and robust assay for PDP 5-HT that will increase throughput for clinical trials.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>19520623</pmid><doi>10.1016/j.jchromb.2009.05.045</doi><tpages>5</tpages></addata></record> |
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| ispartof | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2009-07, Vol.877 (22), p.2163-2167 |
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| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Analysis Analytical, structural and metabolic biochemistry Biological and medical sciences Blood Platelets - metabolism Chromatography, Liquid - methods Functional gastrointestinal disorders Fundamental and applied biological sciences. Psychology Gastrointestinal Diseases - diagnosis Gastrointestinal Diseases - metabolism General pharmacology Humans LC–MS/MS Medical sciences Pharmacology. Drug treatments Protein precipitation Serotonin Serotonin - blood Serotonin - chemistry Serotonin - metabolism Tandem Mass Spectrometry - methods |
| title | Measurement of serotonin in platelet depleted plasma by liquid chromatography tandem mass spectrometry |
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