Quantitative analysis of gemcitabine triphosphate in human peripheral blood mononuclear cells using weak anion-exchange liquid chromatography coupled with tandem mass spectrometry
Gemcitabine triphosphate (dFdCTP) is a highly active metabolite of gemcitabine. It is formed intra‐cellularly via the phosphorylation of gemcitabine by deoxycytidine kinase. The monitoring of dFdCTP in human peripheral blood mononuclear cells (PBMCs), in addition to plasma concentrations of gemcitab...
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| Veröffentlicht in: | Journal of mass spectrometry. 2006-12, Vol.41 (12), p.1633-1642 |
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| creator | Veltkamp, S. A. Hillebrand, M. J. X. Rosing, H. Jansen, R. S. Wickremsinhe, E. R. Perkins, E. J. Schellens, J. H. M. Beijnen, J. H. |
| description | Gemcitabine triphosphate (dFdCTP) is a highly active metabolite of gemcitabine. It is formed intra‐cellularly via the phosphorylation of gemcitabine by deoxycytidine kinase. The monitoring of dFdCTP in human peripheral blood mononuclear cells (PBMCs), in addition to plasma concentrations of gemcitabine and its metabolite 2′,2′‐difluorodeoxyuridine, is considered very useful in determining pharmacokinetic–pharmacodynamic relationships.
We describe a novel sensitive assay for the quantification of dFdCTP in human PBMCs. The method is based on weak anion‐exchange liquid chromatography and detection with tandem mass spectrometry (LC‐MS/MS). The assay has been validated from 1 ng/ml (lower limit of quantification, LLOQ) to 25 ng/ml (upper limit of quantification, ULOQ) using 180 µl aliquots of PBMC extracts containing ∼0.648 mg protein or 3.8 × 106 lysed PBMCs. The LLOQ is equivalent to 94 fmol/106 cells (1 ng/ml = 0.18 ng/180 µl or 0.18 ng/0.648 mg protein = 0.047 ng/106 cells or 94 fmol/106 cells). This highly sensitive assay is capable of quantifying about 200‐fold lower concentrations of dFdCTP in human PBMCs than currently available methods. Copyright © 2006 John Wiley & Sons, Ltd. |
| doi_str_mv | 10.1002/jms.1133 |
| format | Article |
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We describe a novel sensitive assay for the quantification of dFdCTP in human PBMCs. The method is based on weak anion‐exchange liquid chromatography and detection with tandem mass spectrometry (LC‐MS/MS). The assay has been validated from 1 ng/ml (lower limit of quantification, LLOQ) to 25 ng/ml (upper limit of quantification, ULOQ) using 180 µl aliquots of PBMC extracts containing ∼0.648 mg protein or 3.8 × 106 lysed PBMCs. The LLOQ is equivalent to 94 fmol/106 cells (1 ng/ml = 0.18 ng/180 µl or 0.18 ng/0.648 mg protein = 0.047 ng/106 cells or 94 fmol/106 cells). This highly sensitive assay is capable of quantifying about 200‐fold lower concentrations of dFdCTP in human PBMCs than currently available methods. Copyright © 2006 John Wiley & Sons, Ltd.</description><identifier>ISSN: 1076-5174</identifier><identifier>EISSN: 1096-9888</identifier><identifier>DOI: 10.1002/jms.1133</identifier><identifier>PMID: 17117372</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Analysis ; Anions ; Antimetabolites, Antineoplastic - analysis ; Antimetabolites, Antineoplastic - chemistry ; Antimetabolites, Antineoplastic - pharmacokinetics ; Antineoplastic agents ; Biological and medical sciences ; Calibration ; Chromatography, Liquid - methods ; Chromatography, Liquid - standards ; Deoxycytidine - analogs & derivatives ; Deoxycytidine - analysis ; Deoxycytidine - chemistry ; Deoxycytidine - pharmacokinetics ; dFdCTP ; General aspects ; General pharmacology ; Humans ; intra-cellular ; LC-MS/MS ; Leukocyte Count ; Leukocytes, Mononuclear - metabolism ; Medical sciences ; PBMCs ; Pharmacology. Drug treatments ; Proteins - analysis ; Reproducibility of Results ; Sensitivity and Specificity ; Tandem Mass Spectrometry - methods ; Tandem Mass Spectrometry - standards ; validation</subject><ispartof>Journal of mass spectrometry., 2006-12, Vol.41 (12), p.1633-1642</ispartof><rights>Copyright © 2006 John Wiley & Sons, Ltd.</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4183-796835a2c13800fb24210213c628e466dd7f6c0e7af6d1b0e749dc57fcc70f583</citedby><cites>FETCH-LOGICAL-c4183-796835a2c13800fb24210213c628e466dd7f6c0e7af6d1b0e749dc57fcc70f583</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fjms.1133$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjms.1133$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27903,27904,45553,45554</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18348832$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17117372$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Veltkamp, S. A.</creatorcontrib><creatorcontrib>Hillebrand, M. J. X.</creatorcontrib><creatorcontrib>Rosing, H.</creatorcontrib><creatorcontrib>Jansen, R. S.</creatorcontrib><creatorcontrib>Wickremsinhe, E. R.</creatorcontrib><creatorcontrib>Perkins, E. J.</creatorcontrib><creatorcontrib>Schellens, J. H. M.</creatorcontrib><creatorcontrib>Beijnen, J. H.</creatorcontrib><title>Quantitative analysis of gemcitabine triphosphate in human peripheral blood mononuclear cells using weak anion-exchange liquid chromatography coupled with tandem mass spectrometry</title><title>Journal of mass spectrometry.</title><addtitle>J. Mass Spectrom</addtitle><description>Gemcitabine triphosphate (dFdCTP) is a highly active metabolite of gemcitabine. It is formed intra‐cellularly via the phosphorylation of gemcitabine by deoxycytidine kinase. The monitoring of dFdCTP in human peripheral blood mononuclear cells (PBMCs), in addition to plasma concentrations of gemcitabine and its metabolite 2′,2′‐difluorodeoxyuridine, is considered very useful in determining pharmacokinetic–pharmacodynamic relationships.
We describe a novel sensitive assay for the quantification of dFdCTP in human PBMCs. The method is based on weak anion‐exchange liquid chromatography and detection with tandem mass spectrometry (LC‐MS/MS). The assay has been validated from 1 ng/ml (lower limit of quantification, LLOQ) to 25 ng/ml (upper limit of quantification, ULOQ) using 180 µl aliquots of PBMC extracts containing ∼0.648 mg protein or 3.8 × 106 lysed PBMCs. The LLOQ is equivalent to 94 fmol/106 cells (1 ng/ml = 0.18 ng/180 µl or 0.18 ng/0.648 mg protein = 0.047 ng/106 cells or 94 fmol/106 cells). This highly sensitive assay is capable of quantifying about 200‐fold lower concentrations of dFdCTP in human PBMCs than currently available methods. Copyright © 2006 John Wiley & Sons, Ltd.</description><subject>Analysis</subject><subject>Anions</subject><subject>Antimetabolites, Antineoplastic - analysis</subject><subject>Antimetabolites, Antineoplastic - chemistry</subject><subject>Antimetabolites, Antineoplastic - pharmacokinetics</subject><subject>Antineoplastic agents</subject><subject>Biological and medical sciences</subject><subject>Calibration</subject><subject>Chromatography, Liquid - methods</subject><subject>Chromatography, Liquid - standards</subject><subject>Deoxycytidine - analogs & derivatives</subject><subject>Deoxycytidine - analysis</subject><subject>Deoxycytidine - chemistry</subject><subject>Deoxycytidine - pharmacokinetics</subject><subject>dFdCTP</subject><subject>General aspects</subject><subject>General pharmacology</subject><subject>Humans</subject><subject>intra-cellular</subject><subject>LC-MS/MS</subject><subject>Leukocyte Count</subject><subject>Leukocytes, Mononuclear - metabolism</subject><subject>Medical sciences</subject><subject>PBMCs</subject><subject>Pharmacology. Drug treatments</subject><subject>Proteins - analysis</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Tandem Mass Spectrometry - methods</subject><subject>Tandem Mass Spectrometry - standards</subject><subject>validation</subject><issn>1076-5174</issn><issn>1096-9888</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10c1u1DAUBeAIgWgpSDwB8gbEJsWOM4mzRKP-UBUQAgQ768a5mbh17NR2mOa5-oIkmoiuWPnK-nyPrJMkrxk9ZZRmH276cMoY50-SY0arIq2EEE-XuSzSDSvzo-RFCDeU0qrKi-fJESsZK3mZHScP30awUUeI-g8SsGCmoANxLdlhr-b7Wlsk0euhc2HoICLRlnRjD5YMuFyjB0Nq41xDemedHZVB8EShMYGMQdsd2SPczru1syneqw7sDonRd6NuiOq86yG6nYehm4hy42CwIXsdOxLBNtiTHkIgYUAVZ4rRTy-TZy2YgK_W8yT5eX72Y3uZXn-9-LT9eJ2qnAmellUh-AYyxbigtK2zPGM0Y1wVmcC8KJqmbAtFsYS2aFg9D3nVqE3ZKlXSdiP4SfLusHfw7m7EEGWvw_IvsOjGIFmVzwHVAt8foPIuBI-tHLzuwU-SUbkUJOeC5FLQTN-sO8e6x-YRro3M4O0KICgwrQerdHh0gudC8MWlB7fXBqf_Bsqrz9_X4NXrEPH-nwd_K4s5eCN_fbmQ299XojqfX1X8L2TGu2Q</recordid><startdate>200612</startdate><enddate>200612</enddate><creator>Veltkamp, S. A.</creator><creator>Hillebrand, M. J. X.</creator><creator>Rosing, H.</creator><creator>Jansen, R. S.</creator><creator>Wickremsinhe, E. R.</creator><creator>Perkins, E. J.</creator><creator>Schellens, J. H. M.</creator><creator>Beijnen, J. H.</creator><general>John Wiley & Sons, Ltd</general><general>Wiley</general><scope>BSCLL</scope><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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>200612</creationdate><title>Quantitative analysis of gemcitabine triphosphate in human peripheral blood mononuclear cells using weak anion-exchange liquid chromatography coupled with tandem mass spectrometry</title><author>Veltkamp, S. A. ; Hillebrand, M. J. X. ; Rosing, H. ; Jansen, R. S. ; Wickremsinhe, E. R. ; Perkins, E. J. ; Schellens, J. H. M. ; Beijnen, J. H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4183-796835a2c13800fb24210213c628e466dd7f6c0e7af6d1b0e749dc57fcc70f583</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Analysis</topic><topic>Anions</topic><topic>Antimetabolites, Antineoplastic - analysis</topic><topic>Antimetabolites, Antineoplastic - chemistry</topic><topic>Antimetabolites, Antineoplastic - pharmacokinetics</topic><topic>Antineoplastic agents</topic><topic>Biological and medical sciences</topic><topic>Calibration</topic><topic>Chromatography, Liquid - methods</topic><topic>Chromatography, Liquid - standards</topic><topic>Deoxycytidine - analogs & derivatives</topic><topic>Deoxycytidine - analysis</topic><topic>Deoxycytidine - chemistry</topic><topic>Deoxycytidine - pharmacokinetics</topic><topic>dFdCTP</topic><topic>General aspects</topic><topic>General pharmacology</topic><topic>Humans</topic><topic>intra-cellular</topic><topic>LC-MS/MS</topic><topic>Leukocyte Count</topic><topic>Leukocytes, Mononuclear - metabolism</topic><topic>Medical sciences</topic><topic>PBMCs</topic><topic>Pharmacology. Drug treatments</topic><topic>Proteins - analysis</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Tandem Mass Spectrometry - methods</topic><topic>Tandem Mass Spectrometry - standards</topic><topic>validation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Veltkamp, S. A.</creatorcontrib><creatorcontrib>Hillebrand, M. J. X.</creatorcontrib><creatorcontrib>Rosing, H.</creatorcontrib><creatorcontrib>Jansen, R. S.</creatorcontrib><creatorcontrib>Wickremsinhe, E. R.</creatorcontrib><creatorcontrib>Perkins, E. J.</creatorcontrib><creatorcontrib>Schellens, J. H. M.</creatorcontrib><creatorcontrib>Beijnen, J. H.</creatorcontrib><collection>Istex</collection><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>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of mass spectrometry.</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Veltkamp, S. A.</au><au>Hillebrand, M. J. X.</au><au>Rosing, H.</au><au>Jansen, R. S.</au><au>Wickremsinhe, E. R.</au><au>Perkins, E. J.</au><au>Schellens, J. H. M.</au><au>Beijnen, J. H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitative analysis of gemcitabine triphosphate in human peripheral blood mononuclear cells using weak anion-exchange liquid chromatography coupled with tandem mass spectrometry</atitle><jtitle>Journal of mass spectrometry.</jtitle><addtitle>J. Mass Spectrom</addtitle><date>2006-12</date><risdate>2006</risdate><volume>41</volume><issue>12</issue><spage>1633</spage><epage>1642</epage><pages>1633-1642</pages><issn>1076-5174</issn><eissn>1096-9888</eissn><abstract>Gemcitabine triphosphate (dFdCTP) is a highly active metabolite of gemcitabine. It is formed intra‐cellularly via the phosphorylation of gemcitabine by deoxycytidine kinase. The monitoring of dFdCTP in human peripheral blood mononuclear cells (PBMCs), in addition to plasma concentrations of gemcitabine and its metabolite 2′,2′‐difluorodeoxyuridine, is considered very useful in determining pharmacokinetic–pharmacodynamic relationships.
We describe a novel sensitive assay for the quantification of dFdCTP in human PBMCs. The method is based on weak anion‐exchange liquid chromatography and detection with tandem mass spectrometry (LC‐MS/MS). The assay has been validated from 1 ng/ml (lower limit of quantification, LLOQ) to 25 ng/ml (upper limit of quantification, ULOQ) using 180 µl aliquots of PBMC extracts containing ∼0.648 mg protein or 3.8 × 106 lysed PBMCs. The LLOQ is equivalent to 94 fmol/106 cells (1 ng/ml = 0.18 ng/180 µl or 0.18 ng/0.648 mg protein = 0.047 ng/106 cells or 94 fmol/106 cells). This highly sensitive assay is capable of quantifying about 200‐fold lower concentrations of dFdCTP in human PBMCs than currently available methods. Copyright © 2006 John Wiley & Sons, Ltd.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>17117372</pmid><doi>10.1002/jms.1133</doi><tpages>10</tpages></addata></record> |
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| subjects | Analysis Anions Antimetabolites, Antineoplastic - analysis Antimetabolites, Antineoplastic - chemistry Antimetabolites, Antineoplastic - pharmacokinetics Antineoplastic agents Biological and medical sciences Calibration Chromatography, Liquid - methods Chromatography, Liquid - standards Deoxycytidine - analogs & derivatives Deoxycytidine - analysis Deoxycytidine - chemistry Deoxycytidine - pharmacokinetics dFdCTP General aspects General pharmacology Humans intra-cellular LC-MS/MS Leukocyte Count Leukocytes, Mononuclear - metabolism Medical sciences PBMCs Pharmacology. Drug treatments Proteins - analysis Reproducibility of Results Sensitivity and Specificity Tandem Mass Spectrometry - methods Tandem Mass Spectrometry - standards validation |
| title | Quantitative analysis of gemcitabine triphosphate in human peripheral blood mononuclear cells using weak anion-exchange liquid chromatography coupled with tandem mass spectrometry |
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