Quantification of antiretroviral drugs in dried blood spot samples by means of liquid chromatography/tandem mass spectrometry
For the first time approved antiretroviral drugs, i.e. protease inhibitors (PI) and non‐nucleoside reverse transcriptase inhibitors (NNRTI), were quantified in dried blood spots (DBS) from HIV/AIDS patient whole blood samples as the basis for therapeutic drug monitoring (TDM) by a robust simultaneou...
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description | For the first time approved antiretroviral drugs, i.e. protease inhibitors (PI) and non‐nucleoside reverse transcriptase inhibitors (NNRTI), were quantified in dried blood spots (DBS) from HIV/AIDS patient whole blood samples as the basis for therapeutic drug monitoring (TDM) by a robust simultaneous liquid chromatography/tandem mass spectrometry (LC/MS/MS) method. This study included seven PI (amprenavir, nelfinavir, indinavir, lopinavir, saquinavir, ritonavir, atazanavir) and two NNRTI (nevirapine, efavirenz). LC/MS/MS coupling was realized using a Phenomenex® Synergy Max RP LC column (150 × 2 mm, 4 µ) in combination with a tandem mass spectrometer (API 2000, Applied Biosystems/MDS Sciex Concord) operating in positive and negative multiple reaction monitoring (MRM) mode with reserpine as internal standard. DBS samples were punched out and extracted with 50:50 MeOH/0.2 M ZnSO4 (v/v) as extraction reagent. The method performance data for the drugs in DBS like limits of detection (LOD, 8–70 ng/mL), lower limits of quantification (LLOQ, 41–102 ng/mL), linearity (R2, 0.9981–0.9999), linear concentration ranges (41–10.000 ng/mL), accuracies (92–113%), recoveries (62–94%), and ion suppression were investigated and are comparable to data obtained from human plasma, which is the current standard matrix for TDM of PI and NNRTI. In this case, off‐line plasma sample preparation was performed by means of simple protein precipitation with 80:20 methanol/0.2 M ZnSO4 (v/v) as precipitation reagent. Significant correlations between real patient plasma and DBS were obtained for samples containing lopinavir, atazanavir, ritonavir, saquinavir, and efavirenz. DBS preparation as sampling alternative is well suited and practicable for TDM minimizing the high infection risk of HIV/AIDS samples and may facilitate sample mailing. Copyright © 2005 John Wiley & Sons, Ltd. |
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This study included seven PI (amprenavir, nelfinavir, indinavir, lopinavir, saquinavir, ritonavir, atazanavir) and two NNRTI (nevirapine, efavirenz). LC/MS/MS coupling was realized using a Phenomenex® Synergy Max RP LC column (150 × 2 mm, 4 µ) in combination with a tandem mass spectrometer (API 2000, Applied Biosystems/MDS Sciex Concord) operating in positive and negative multiple reaction monitoring (MRM) mode with reserpine as internal standard. DBS samples were punched out and extracted with 50:50 MeOH/0.2 M ZnSO4 (v/v) as extraction reagent. The method performance data for the drugs in DBS like limits of detection (LOD, 8–70 ng/mL), lower limits of quantification (LLOQ, 41–102 ng/mL), linearity (R2, 0.9981–0.9999), linear concentration ranges (41–10.000 ng/mL), accuracies (92–113%), recoveries (62–94%), and ion suppression were investigated and are comparable to data obtained from human plasma, which is the current standard matrix for TDM of PI and NNRTI. In this case, off‐line plasma sample preparation was performed by means of simple protein precipitation with 80:20 methanol/0.2 M ZnSO4 (v/v) as precipitation reagent. Significant correlations between real patient plasma and DBS were obtained for samples containing lopinavir, atazanavir, ritonavir, saquinavir, and efavirenz. DBS preparation as sampling alternative is well suited and practicable for TDM minimizing the high infection risk of HIV/AIDS samples and may facilitate sample mailing. Copyright © 2005 John Wiley & Sons, Ltd.</description><identifier>ISSN: 0951-4198</identifier><identifier>EISSN: 1097-0231</identifier><identifier>DOI: 10.1002/rcm.2158</identifier><identifier>PMID: 16193530</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Acquired Immunodeficiency Syndrome - blood ; Blood Specimen Collection - methods ; Chromatography, High Pressure Liquid - methods ; Drug Monitoring - methods ; HIV Protease Inhibitors - blood ; Humans ; Reproducibility of Results ; Reverse Transcriptase Inhibitors - blood ; Spectrometry, Mass, Electrospray Ionization - methods</subject><ispartof>Rapid communications in mass spectrometry, 2005-01, Vol.19 (21), p.2995-3001</ispartof><rights>Copyright © 2005 John Wiley & Sons, Ltd.</rights><rights>(c) 2005 John Wiley & Sons, Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4578-97853ddbd0740c725c5b323e15d6c3aae18c22502219aa6e9214cf13107a18d73</citedby><cites>FETCH-LOGICAL-c4578-97853ddbd0740c725c5b323e15d6c3aae18c22502219aa6e9214cf13107a18d73</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%2Frcm.2158$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Frcm.2158$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16193530$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Koal, Therese</creatorcontrib><creatorcontrib>Burhenne, Heike</creatorcontrib><creatorcontrib>Römling, Regina</creatorcontrib><creatorcontrib>Svoboda, Michal</creatorcontrib><creatorcontrib>Resch, Klaus</creatorcontrib><creatorcontrib>Kaever, Volkhard</creatorcontrib><title>Quantification of antiretroviral drugs in dried blood spot samples by means of liquid chromatography/tandem mass spectrometry</title><title>Rapid communications in mass spectrometry</title><addtitle>Rapid Commun. Mass Spectrom</addtitle><description>For the first time approved antiretroviral drugs, i.e. protease inhibitors (PI) and non‐nucleoside reverse transcriptase inhibitors (NNRTI), were quantified in dried blood spots (DBS) from HIV/AIDS patient whole blood samples as the basis for therapeutic drug monitoring (TDM) by a robust simultaneous liquid chromatography/tandem mass spectrometry (LC/MS/MS) method. This study included seven PI (amprenavir, nelfinavir, indinavir, lopinavir, saquinavir, ritonavir, atazanavir) and two NNRTI (nevirapine, efavirenz). LC/MS/MS coupling was realized using a Phenomenex® Synergy Max RP LC column (150 × 2 mm, 4 µ) in combination with a tandem mass spectrometer (API 2000, Applied Biosystems/MDS Sciex Concord) operating in positive and negative multiple reaction monitoring (MRM) mode with reserpine as internal standard. DBS samples were punched out and extracted with 50:50 MeOH/0.2 M ZnSO4 (v/v) as extraction reagent. The method performance data for the drugs in DBS like limits of detection (LOD, 8–70 ng/mL), lower limits of quantification (LLOQ, 41–102 ng/mL), linearity (R2, 0.9981–0.9999), linear concentration ranges (41–10.000 ng/mL), accuracies (92–113%), recoveries (62–94%), and ion suppression were investigated and are comparable to data obtained from human plasma, which is the current standard matrix for TDM of PI and NNRTI. In this case, off‐line plasma sample preparation was performed by means of simple protein precipitation with 80:20 methanol/0.2 M ZnSO4 (v/v) as precipitation reagent. Significant correlations between real patient plasma and DBS were obtained for samples containing lopinavir, atazanavir, ritonavir, saquinavir, and efavirenz. DBS preparation as sampling alternative is well suited and practicable for TDM minimizing the high infection risk of HIV/AIDS samples and may facilitate sample mailing. Copyright © 2005 John Wiley & Sons, Ltd.</description><subject>Acquired Immunodeficiency Syndrome - blood</subject><subject>Blood Specimen Collection - methods</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Drug Monitoring - methods</subject><subject>HIV Protease Inhibitors - blood</subject><subject>Humans</subject><subject>Reproducibility of Results</subject><subject>Reverse Transcriptase Inhibitors - blood</subject><subject>Spectrometry, Mass, Electrospray Ionization - methods</subject><issn>0951-4198</issn><issn>1097-0231</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kMFu1DAQhi0EokuLxBMgnxCXtB47jpMjWqAglqJWhUpcLMd2WkMcp3YC5MC749VGcEJzmBnpm2-kH6FnQE6BEHoWtT-lwOsHaAOkEQWhDB6iDWk4FCU09RF6ktI3QgA4JY_REVTQMM7IBv2-nNUwuc5pNbkw4NDh_R7tFMMPF1WPTZxvE3ZDHpw1uO1DMDiNYcJJ-bG3CbcL9lYNaX_cu_vZGazvYvBqCrdRjXfL2aQGYz32KqV8anWW-_xhOUGPOtUn-3Ttx-jz2zfX23fF7tP5--2rXaFLLuqiETVnxrSGiJJoQbnmLaPMAjeVZkpZqDWlnFAKjVKVbSiUugMGRCiojWDH6MXBO8ZwP9s0Se-Stn2vBhvmJKs6FyvLDL48gDqGlKLt5BidV3GRQOQ-apmjlvuoM_p8dc6tt-YfuGabgeIA_HS9Xf4rklfbj6tw5V2a7K-_vIrfZSWY4PLm4lxe7L5uX998qeQH9gfwx5lz</recordid><startdate>20050101</startdate><enddate>20050101</enddate><creator>Koal, Therese</creator><creator>Burhenne, Heike</creator><creator>Römling, Regina</creator><creator>Svoboda, Michal</creator><creator>Resch, Klaus</creator><creator>Kaever, Volkhard</creator><general>John Wiley & Sons, Ltd</general><scope>BSCLL</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>7X8</scope></search><sort><creationdate>20050101</creationdate><title>Quantification of antiretroviral drugs in dried blood spot samples by means of liquid chromatography/tandem mass spectrometry</title><author>Koal, Therese ; Burhenne, Heike ; Römling, Regina ; Svoboda, Michal ; Resch, Klaus ; Kaever, Volkhard</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4578-97853ddbd0740c725c5b323e15d6c3aae18c22502219aa6e9214cf13107a18d73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Acquired Immunodeficiency Syndrome - blood</topic><topic>Blood Specimen Collection - methods</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Drug Monitoring - methods</topic><topic>HIV Protease Inhibitors - blood</topic><topic>Humans</topic><topic>Reproducibility of Results</topic><topic>Reverse Transcriptase Inhibitors - blood</topic><topic>Spectrometry, Mass, Electrospray Ionization - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Koal, Therese</creatorcontrib><creatorcontrib>Burhenne, Heike</creatorcontrib><creatorcontrib>Römling, Regina</creatorcontrib><creatorcontrib>Svoboda, Michal</creatorcontrib><creatorcontrib>Resch, Klaus</creatorcontrib><creatorcontrib>Kaever, Volkhard</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Rapid communications in mass spectrometry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Koal, Therese</au><au>Burhenne, Heike</au><au>Römling, Regina</au><au>Svoboda, Michal</au><au>Resch, Klaus</au><au>Kaever, Volkhard</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantification of antiretroviral drugs in dried blood spot samples by means of liquid chromatography/tandem mass spectrometry</atitle><jtitle>Rapid communications in mass spectrometry</jtitle><addtitle>Rapid Commun. Mass Spectrom</addtitle><date>2005-01-01</date><risdate>2005</risdate><volume>19</volume><issue>21</issue><spage>2995</spage><epage>3001</epage><pages>2995-3001</pages><issn>0951-4198</issn><eissn>1097-0231</eissn><abstract>For the first time approved antiretroviral drugs, i.e. protease inhibitors (PI) and non‐nucleoside reverse transcriptase inhibitors (NNRTI), were quantified in dried blood spots (DBS) from HIV/AIDS patient whole blood samples as the basis for therapeutic drug monitoring (TDM) by a robust simultaneous liquid chromatography/tandem mass spectrometry (LC/MS/MS) method. This study included seven PI (amprenavir, nelfinavir, indinavir, lopinavir, saquinavir, ritonavir, atazanavir) and two NNRTI (nevirapine, efavirenz). LC/MS/MS coupling was realized using a Phenomenex® Synergy Max RP LC column (150 × 2 mm, 4 µ) in combination with a tandem mass spectrometer (API 2000, Applied Biosystems/MDS Sciex Concord) operating in positive and negative multiple reaction monitoring (MRM) mode with reserpine as internal standard. DBS samples were punched out and extracted with 50:50 MeOH/0.2 M ZnSO4 (v/v) as extraction reagent. The method performance data for the drugs in DBS like limits of detection (LOD, 8–70 ng/mL), lower limits of quantification (LLOQ, 41–102 ng/mL), linearity (R2, 0.9981–0.9999), linear concentration ranges (41–10.000 ng/mL), accuracies (92–113%), recoveries (62–94%), and ion suppression were investigated and are comparable to data obtained from human plasma, which is the current standard matrix for TDM of PI and NNRTI. In this case, off‐line plasma sample preparation was performed by means of simple protein precipitation with 80:20 methanol/0.2 M ZnSO4 (v/v) as precipitation reagent. Significant correlations between real patient plasma and DBS were obtained for samples containing lopinavir, atazanavir, ritonavir, saquinavir, and efavirenz. DBS preparation as sampling alternative is well suited and practicable for TDM minimizing the high infection risk of HIV/AIDS samples and may facilitate sample mailing. Copyright © 2005 John Wiley & Sons, Ltd.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>16193530</pmid><doi>10.1002/rcm.2158</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Acquired Immunodeficiency Syndrome - blood Blood Specimen Collection - methods Chromatography, High Pressure Liquid - methods Drug Monitoring - methods HIV Protease Inhibitors - blood Humans Reproducibility of Results Reverse Transcriptase Inhibitors - blood Spectrometry, Mass, Electrospray Ionization - methods |
title | Quantification of antiretroviral drugs in dried blood spot samples by means of liquid chromatography/tandem mass spectrometry |
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