On-Line HPLC Electrospray Mass Spectrometry of Phosphorothioate Oligonucleotide Metabolites

Metabolism of 2‘-deoxyphosphorothioate oligonucleotides ISIS 11061 and ISIS 11637 was examined with capillary gel electrophoresis (CGE) and on-line HPLC electrospray mass spectrometry (HPLC/ES-MS). Oligonucleotides were isolated from plasma, liver, and kidneys of rats injected with ISIS 11061 and IS...

Full description

Saved in:
Bibliographic Details
Published in:Analytical chemistry (Washington) 1997-02, Vol.69 (3), p.313-319
Main Authors: Gaus, Hans J, Owens, Stephen R, Winniman, Michael, Cooper, Scott, Cummins, Lendell L
Format: Article
Language:English
Subjects:
Analysis
Animals
Biochemistry
Biological and medical sciences
Chromatography, High Pressure Liquid
Drugs
General pharmacology
Mass Spectrometry
Medical sciences
Metabolism
Oligonucleotides - analysis
Oligonucleotides - metabolism
Pharmacology. Drug treatments
Rats
Scientific imaging
Thionucleotides - analysis
Thionucleotides - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
cited_by cdi_FETCH-LOGICAL-a435t-8abe74d1a293acd5d5217c62c46fffbad89d39cb37f7719c5ab10fe5bb81c2ea3
cites cdi_FETCH-LOGICAL-a435t-8abe74d1a293acd5d5217c62c46fffbad89d39cb37f7719c5ab10fe5bb81c2ea3
container_end_page 319
container_issue 3
container_start_page 313
container_title Analytical chemistry (Washington)
container_volume 69
creator Gaus, Hans J
Owens, Stephen R
Winniman, Michael
Cooper, Scott
Cummins, Lendell L
description Metabolism of 2‘-deoxyphosphorothioate oligonucleotides ISIS 11061 and ISIS 11637 was examined with capillary gel electrophoresis (CGE) and on-line HPLC electrospray mass spectrometry (HPLC/ES-MS). Oligonucleotides were isolated from plasma, liver, and kidneys of rats injected with ISIS 11061 and ISIS 11637. Metabolites found in plasma were consistent with 3‘-exonuclease activity. Metabolites isolated from liver and kidney were consistent with 3‘- and/or 5‘-exonuclease activity. HPLC/ES-MS analysis of ISIS 11061 isolated from kidney indicated extensive degradation from the 3‘ terminus, but metabolites consistent with 5‘ degradation and combinations of 3‘ and 5‘ truncations also were observed. ISIS 11061 isolated from liver showed less extensive degradation. The 5‘ truncated metabolites represented the predominant species in contrast to the kidney sample. Metabolites with masses consistent with combinations of 3‘ and 5‘ truncations were also observed in liver. The metabolic profiles generated by CGE analysis of these samples agreed qualitatively with mass spectrometric results. HPLC/ES-MS enabled the simultaneous determination of degradation products that are the same length but differ in composition. CGE could discriminate species that differed by one nucleotide in length. HPLC/ES-MS was shown to be a useful tool to study the complex metabolism of antisense oligonucleotides in vivo.
doi_str_mv 10.1021/ac960557q
format article
fullrecord <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_78823511</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>11160255</sourcerecordid><originalsourceid>FETCH-LOGICAL-a435t-8abe74d1a293acd5d5217c62c46fffbad89d39cb37f7719c5ab10fe5bb81c2ea3</originalsourceid><addsrcrecordid>eNqF0c9v0zAUB3ALgUYZHPgDkCIESBwCz3Ycx0dUbWxSuxatiAMH68VxaEYad7Yjrf893loVaRw4WfL3o6f3g5DXFD5RYPQzGlWCEPL2CZlQwSAvq4o9JRMA4DmTAM_JixBuACgFWp6QEwUcoKgm5OdiyGfdYLOL5WyanfXWRO_C1uMum2MI2fX24Wdjo99lrs2W65SunXdx3TmMNlv03S83jKa3LnaNzeY2Yu36LtrwkjxrsQ_21eE9Jd_Pz1bTi3y2-Ho5_TLLseAi5hXWVhYNRaY4mkY0glFpSmaKsm3bGptKNVyZmstWSqqMwJpCa0VdV9Qwi_yUfNjX3Xp3O9oQ9aYLxvY9DtaNQcu0DC4o_S-kQhUV5WWCbx_BGzf6IQ2hU2-VUKpgCX3cI5M2Frxt9dZ3G_Q7TUHfn0Ufz5Lsm0PBsd7Y5igPd0j5u0OOwWDfehxMF46MiRLkQ1_5nnUh2rtjjP63LiWXQq-W13q-VD-uvq2khuTf7z2a8HeEf9v7AwtFsEc</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>217859942</pqid></control><display><type>article</type><title>On-Line HPLC Electrospray Mass Spectrometry of Phosphorothioate Oligonucleotide Metabolites</title><source>American Chemical Society:Jisc Collections:American Chemical Society Read &amp; Publish Agreement 2022-2024 (Reading list)</source><creator>Gaus, Hans J ; Owens, Stephen R ; Winniman, Michael ; Cooper, Scott ; Cummins, Lendell L</creator><creatorcontrib>Gaus, Hans J ; Owens, Stephen R ; Winniman, Michael ; Cooper, Scott ; Cummins, Lendell L</creatorcontrib><description>Metabolism of 2‘-deoxyphosphorothioate oligonucleotides ISIS 11061 and ISIS 11637 was examined with capillary gel electrophoresis (CGE) and on-line HPLC electrospray mass spectrometry (HPLC/ES-MS). Oligonucleotides were isolated from plasma, liver, and kidneys of rats injected with ISIS 11061 and ISIS 11637. Metabolites found in plasma were consistent with 3‘-exonuclease activity. Metabolites isolated from liver and kidney were consistent with 3‘- and/or 5‘-exonuclease activity. HPLC/ES-MS analysis of ISIS 11061 isolated from kidney indicated extensive degradation from the 3‘ terminus, but metabolites consistent with 5‘ degradation and combinations of 3‘ and 5‘ truncations also were observed. ISIS 11061 isolated from liver showed less extensive degradation. The 5‘ truncated metabolites represented the predominant species in contrast to the kidney sample. Metabolites with masses consistent with combinations of 3‘ and 5‘ truncations were also observed in liver. The metabolic profiles generated by CGE analysis of these samples agreed qualitatively with mass spectrometric results. HPLC/ES-MS enabled the simultaneous determination of degradation products that are the same length but differ in composition. CGE could discriminate species that differed by one nucleotide in length. HPLC/ES-MS was shown to be a useful tool to study the complex metabolism of antisense oligonucleotides in vivo.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac960557q</identifier><identifier>PMID: 9030048</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Analysis ; Animals ; Biochemistry ; Biological and medical sciences ; Chromatography, High Pressure Liquid ; Drugs ; General pharmacology ; Mass Spectrometry ; Medical sciences ; Metabolism ; Oligonucleotides - analysis ; Oligonucleotides - metabolism ; Pharmacology. Drug treatments ; Rats ; Scientific imaging ; Thionucleotides - analysis ; Thionucleotides - metabolism</subject><ispartof>Analytical chemistry (Washington), 1997-02, Vol.69 (3), p.313-319</ispartof><rights>Copyright © 1997 American Chemical Society</rights><rights>1997 INIST-CNRS</rights><rights>Copyright American Chemical Society Feb 1, 1997</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a435t-8abe74d1a293acd5d5217c62c46fffbad89d39cb37f7719c5ab10fe5bb81c2ea3</citedby><cites>FETCH-LOGICAL-a435t-8abe74d1a293acd5d5217c62c46fffbad89d39cb37f7719c5ab10fe5bb81c2ea3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=2560736$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9030048$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gaus, Hans J</creatorcontrib><creatorcontrib>Owens, Stephen R</creatorcontrib><creatorcontrib>Winniman, Michael</creatorcontrib><creatorcontrib>Cooper, Scott</creatorcontrib><creatorcontrib>Cummins, Lendell L</creatorcontrib><title>On-Line HPLC Electrospray Mass Spectrometry of Phosphorothioate Oligonucleotide Metabolites</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Metabolism of 2‘-deoxyphosphorothioate oligonucleotides ISIS 11061 and ISIS 11637 was examined with capillary gel electrophoresis (CGE) and on-line HPLC electrospray mass spectrometry (HPLC/ES-MS). Oligonucleotides were isolated from plasma, liver, and kidneys of rats injected with ISIS 11061 and ISIS 11637. Metabolites found in plasma were consistent with 3‘-exonuclease activity. Metabolites isolated from liver and kidney were consistent with 3‘- and/or 5‘-exonuclease activity. HPLC/ES-MS analysis of ISIS 11061 isolated from kidney indicated extensive degradation from the 3‘ terminus, but metabolites consistent with 5‘ degradation and combinations of 3‘ and 5‘ truncations also were observed. ISIS 11061 isolated from liver showed less extensive degradation. The 5‘ truncated metabolites represented the predominant species in contrast to the kidney sample. Metabolites with masses consistent with combinations of 3‘ and 5‘ truncations were also observed in liver. The metabolic profiles generated by CGE analysis of these samples agreed qualitatively with mass spectrometric results. HPLC/ES-MS enabled the simultaneous determination of degradation products that are the same length but differ in composition. CGE could discriminate species that differed by one nucleotide in length. HPLC/ES-MS was shown to be a useful tool to study the complex metabolism of antisense oligonucleotides in vivo.</description><subject>Analysis</subject><subject>Animals</subject><subject>Biochemistry</subject><subject>Biological and medical sciences</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Drugs</subject><subject>General pharmacology</subject><subject>Mass Spectrometry</subject><subject>Medical sciences</subject><subject>Metabolism</subject><subject>Oligonucleotides - analysis</subject><subject>Oligonucleotides - metabolism</subject><subject>Pharmacology. Drug treatments</subject><subject>Rats</subject><subject>Scientific imaging</subject><subject>Thionucleotides - analysis</subject><subject>Thionucleotides - metabolism</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><recordid>eNqF0c9v0zAUB3ALgUYZHPgDkCIESBwCz3Ycx0dUbWxSuxatiAMH68VxaEYad7Yjrf893loVaRw4WfL3o6f3g5DXFD5RYPQzGlWCEPL2CZlQwSAvq4o9JRMA4DmTAM_JixBuACgFWp6QEwUcoKgm5OdiyGfdYLOL5WyanfXWRO_C1uMum2MI2fX24Wdjo99lrs2W65SunXdx3TmMNlv03S83jKa3LnaNzeY2Yu36LtrwkjxrsQ_21eE9Jd_Pz1bTi3y2-Ho5_TLLseAi5hXWVhYNRaY4mkY0glFpSmaKsm3bGptKNVyZmstWSqqMwJpCa0VdV9Qwi_yUfNjX3Xp3O9oQ9aYLxvY9DtaNQcu0DC4o_S-kQhUV5WWCbx_BGzf6IQ2hU2-VUKpgCX3cI5M2Frxt9dZ3G_Q7TUHfn0Ufz5Lsm0PBsd7Y5igPd0j5u0OOwWDfehxMF46MiRLkQ1_5nnUh2rtjjP63LiWXQq-W13q-VD-uvq2khuTf7z2a8HeEf9v7AwtFsEc</recordid><startdate>19970201</startdate><enddate>19970201</enddate><creator>Gaus, Hans J</creator><creator>Owens, Stephen R</creator><creator>Winniman, Michael</creator><creator>Cooper, Scott</creator><creator>Cummins, Lendell L</creator><general>American Chemical Society</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19970201</creationdate><title>On-Line HPLC Electrospray Mass Spectrometry of Phosphorothioate Oligonucleotide Metabolites</title><author>Gaus, Hans J ; Owens, Stephen R ; Winniman, Michael ; Cooper, Scott ; Cummins, Lendell L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a435t-8abe74d1a293acd5d5217c62c46fffbad89d39cb37f7719c5ab10fe5bb81c2ea3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Analysis</topic><topic>Animals</topic><topic>Biochemistry</topic><topic>Biological and medical sciences</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Drugs</topic><topic>General pharmacology</topic><topic>Mass Spectrometry</topic><topic>Medical sciences</topic><topic>Metabolism</topic><topic>Oligonucleotides - analysis</topic><topic>Oligonucleotides - metabolism</topic><topic>Pharmacology. Drug treatments</topic><topic>Rats</topic><topic>Scientific imaging</topic><topic>Thionucleotides - analysis</topic><topic>Thionucleotides - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gaus, Hans J</creatorcontrib><creatorcontrib>Owens, Stephen R</creatorcontrib><creatorcontrib>Winniman, Michael</creatorcontrib><creatorcontrib>Cooper, Scott</creatorcontrib><creatorcontrib>Cummins, Lendell L</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>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics &amp; Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical &amp; Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology &amp; Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts – Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gaus, Hans J</au><au>Owens, Stephen R</au><au>Winniman, Michael</au><au>Cooper, Scott</au><au>Cummins, Lendell L</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>On-Line HPLC Electrospray Mass Spectrometry of Phosphorothioate Oligonucleotide Metabolites</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>1997-02-01</date><risdate>1997</risdate><volume>69</volume><issue>3</issue><spage>313</spage><epage>319</epage><pages>313-319</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Metabolism of 2‘-deoxyphosphorothioate oligonucleotides ISIS 11061 and ISIS 11637 was examined with capillary gel electrophoresis (CGE) and on-line HPLC electrospray mass spectrometry (HPLC/ES-MS). Oligonucleotides were isolated from plasma, liver, and kidneys of rats injected with ISIS 11061 and ISIS 11637. Metabolites found in plasma were consistent with 3‘-exonuclease activity. Metabolites isolated from liver and kidney were consistent with 3‘- and/or 5‘-exonuclease activity. HPLC/ES-MS analysis of ISIS 11061 isolated from kidney indicated extensive degradation from the 3‘ terminus, but metabolites consistent with 5‘ degradation and combinations of 3‘ and 5‘ truncations also were observed. ISIS 11061 isolated from liver showed less extensive degradation. The 5‘ truncated metabolites represented the predominant species in contrast to the kidney sample. Metabolites with masses consistent with combinations of 3‘ and 5‘ truncations were also observed in liver. The metabolic profiles generated by CGE analysis of these samples agreed qualitatively with mass spectrometric results. HPLC/ES-MS enabled the simultaneous determination of degradation products that are the same length but differ in composition. CGE could discriminate species that differed by one nucleotide in length. HPLC/ES-MS was shown to be a useful tool to study the complex metabolism of antisense oligonucleotides in vivo.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>9030048</pmid><doi>10.1021/ac960557q</doi><tpages>7</tpages></addata></record>
fulltext fulltext
identifier ISSN: 0003-2700
ispartof Analytical chemistry (Washington), 1997-02, Vol.69 (3), p.313-319
issn 0003-2700
1520-6882
language eng
recordid cdi_proquest_miscellaneous_78823511
source American Chemical Society:Jisc Collections:American Chemical Society Read & Publish Agreement 2022-2024 (Reading list)
subjects Analysis
Animals
Biochemistry
Biological and medical sciences
Chromatography, High Pressure Liquid
Drugs
General pharmacology
Mass Spectrometry
Medical sciences
Metabolism
Oligonucleotides - analysis
Oligonucleotides - metabolism
Pharmacology. Drug treatments
Rats
Scientific imaging
Thionucleotides - analysis
Thionucleotides - metabolism
title On-Line HPLC Electrospray Mass Spectrometry of Phosphorothioate Oligonucleotide Metabolites
url http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T22%3A40%3A55IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=On-Line%20HPLC%20Electrospray%20Mass%20Spectrometry%20of%20Phosphorothioate%20Oligonucleotide%20Metabolites&rft.jtitle=Analytical%20chemistry%20(Washington)&rft.au=Gaus,%20Hans%20J&rft.date=1997-02-01&rft.volume=69&rft.issue=3&rft.spage=313&rft.epage=319&rft.pages=313-319&rft.issn=0003-2700&rft.eissn=1520-6882&rft.coden=ANCHAM&rft_id=info:doi/10.1021/ac960557q&rft_dat=%3Cproquest_cross%3E11160255%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-a435t-8abe74d1a293acd5d5217c62c46fffbad89d39cb37f7719c5ab10fe5bb81c2ea3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=217859942&rft_id=info:pmid/9030048&rfr_iscdi=true