Absolute quantification of superoxide dismutase (SOD) using species-specific isotope dilution analysis
Here we report for the first time the use of species-specific isotope dilution mass spectrometry for the absolute quantification of a metalloprotein using non-denaturing gel electrophoresis laser ablation inductively coupled plasma mass spectrometry (GE-LA-ICP-MS). The concept utilises the intrinsic...
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| Veröffentlicht in: | Analytical and bioanalytical chemistry 2010-08, Vol.397 (8), p.3515-3524 |
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| creator | Deitrich, Christian L Braukmann, Sandra Raab, Andrea Munro, Caroline Pioselli, Barbara Krupp, Eva M Thomas-Oates, Jane E Feldmann, Jörg |
| description | Here we report for the first time the use of species-specific isotope dilution mass spectrometry for the absolute quantification of a metalloprotein using non-denaturing gel electrophoresis laser ablation inductively coupled plasma mass spectrometry (GE-LA-ICP-MS). The concept utilises the intrinsic metals of the metalloprotein for labelling of the isotopically labelled spike (⁶⁵Cu, ⁶⁸Zn SOD). The stability of the metal-protein complex under non-denaturing conditions during 1-D PAGE was confirmed and the performance of the method evaluated. Between 4 and 64 μg, SOD was quantified with a recovery rate between 82% and 110% in a standard. The use of the isotopically enriched SOD was utilised to identify the extent of orthogonal diffusion in 1-D gel electrophoresis. Orthogonal diffusion of natural and isotopically enriched SOD in the gel can interfere with the correct determination of the isotope ratios. The matrix effect of a cytosolic liver extract on the non-covalently bound copper and zinc in SOD was evaluated and no significant metal loss from the SOD spike was observed. This study represents the first step necessary for establishing and evaluating the use of a species-specific isotope dilution approach for the absolute quantification of SOD in real samples based on the combination of gel electrophoresis and LA-ICP-MS. |
| doi_str_mv | 10.1007/s00216-010-3680-1 |
| format | Article |
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The concept utilises the intrinsic metals of the metalloprotein for labelling of the isotopically labelled spike (⁶⁵Cu, ⁶⁸Zn SOD). The stability of the metal-protein complex under non-denaturing conditions during 1-D PAGE was confirmed and the performance of the method evaluated. Between 4 and 64 μg, SOD was quantified with a recovery rate between 82% and 110% in a standard. The use of the isotopically enriched SOD was utilised to identify the extent of orthogonal diffusion in 1-D gel electrophoresis. Orthogonal diffusion of natural and isotopically enriched SOD in the gel can interfere with the correct determination of the isotope ratios. The matrix effect of a cytosolic liver extract on the non-covalently bound copper and zinc in SOD was evaluated and no significant metal loss from the SOD spike was observed. This study represents the first step necessary for establishing and evaluating the use of a species-specific isotope dilution approach for the absolute quantification of SOD in real samples based on the combination of gel electrophoresis and LA-ICP-MS.</description><identifier>ISSN: 1618-2642</identifier><identifier>EISSN: 1618-2650</identifier><identifier>DOI: 10.1007/s00216-010-3680-1</identifier><identifier>PMID: 20393840</identifier><language>eng</language><publisher>Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag</publisher><subject>Affinity labeling ; Analysis ; Analytical Chemistry ; Animals ; Biochemistry ; Cattle ; Characterization and Evaluation of Materials ; Chemistry ; Chemistry and Materials Science ; Chemistry Techniques, Analytical - methods ; Copper - analysis ; Diffusion ; Electrophoresis ; Enrichment ; Food Science ; Gel electrophoresis ; ICP-MS ; Inductively coupled plasma mass spectrometry ; Isotope dilution mass spectrometry ; Isotope dilution techniques ; Isotope Labeling ; Isotopes ; Isotopes - analysis ; Labelling ; Laboratory Medicine ; Laser ablation ; Liver ; Liver - chemistry ; Liver - enzymology ; Mass Spectrometry ; Metalloenzymes ; Methods ; Monitoring/Environmental Analysis ; Original Paper ; Physiological aspects ; Properties ; Protein quantification ; Proteins ; Proteomics ; Separation ; Sod ; Speciation analysis ; Species Specificity ; Spectra ; Spikes ; Superoxide dismutase ; Superoxide Dismutase - chemistry ; Zinc ; Zinc Isotopes - analysis</subject><ispartof>Analytical and bioanalytical chemistry, 2010-08, Vol.397 (8), p.3515-3524</ispartof><rights>Springer-Verlag 2010</rights><rights>COPYRIGHT 2010 Springer</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c565t-ceb64718ebcd4878ccadde63ac3800e511ed135c2fcc3cb7a20de6fc96f8cb233</citedby><cites>FETCH-LOGICAL-c565t-ceb64718ebcd4878ccadde63ac3800e511ed135c2fcc3cb7a20de6fc96f8cb233</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00216-010-3680-1$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00216-010-3680-1$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20393840$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Deitrich, Christian L</creatorcontrib><creatorcontrib>Braukmann, Sandra</creatorcontrib><creatorcontrib>Raab, Andrea</creatorcontrib><creatorcontrib>Munro, Caroline</creatorcontrib><creatorcontrib>Pioselli, Barbara</creatorcontrib><creatorcontrib>Krupp, Eva M</creatorcontrib><creatorcontrib>Thomas-Oates, Jane E</creatorcontrib><creatorcontrib>Feldmann, Jörg</creatorcontrib><title>Absolute quantification of superoxide dismutase (SOD) using species-specific isotope dilution analysis</title><title>Analytical and bioanalytical chemistry</title><addtitle>Anal Bioanal Chem</addtitle><addtitle>Anal Bioanal Chem</addtitle><description>Here we report for the first time the use of species-specific isotope dilution mass spectrometry for the absolute quantification of a metalloprotein using non-denaturing gel electrophoresis laser ablation inductively coupled plasma mass spectrometry (GE-LA-ICP-MS). The concept utilises the intrinsic metals of the metalloprotein for labelling of the isotopically labelled spike (⁶⁵Cu, ⁶⁸Zn SOD). The stability of the metal-protein complex under non-denaturing conditions during 1-D PAGE was confirmed and the performance of the method evaluated. Between 4 and 64 μg, SOD was quantified with a recovery rate between 82% and 110% in a standard. The use of the isotopically enriched SOD was utilised to identify the extent of orthogonal diffusion in 1-D gel electrophoresis. Orthogonal diffusion of natural and isotopically enriched SOD in the gel can interfere with the correct determination of the isotope ratios. The matrix effect of a cytosolic liver extract on the non-covalently bound copper and zinc in SOD was evaluated and no significant metal loss from the SOD spike was observed. This study represents the first step necessary for establishing and evaluating the use of a species-specific isotope dilution approach for the absolute quantification of SOD in real samples based on the combination of gel electrophoresis and LA-ICP-MS.</description><subject>Affinity labeling</subject><subject>Analysis</subject><subject>Analytical Chemistry</subject><subject>Animals</subject><subject>Biochemistry</subject><subject>Cattle</subject><subject>Characterization and Evaluation of Materials</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Chemistry Techniques, Analytical - methods</subject><subject>Copper - analysis</subject><subject>Diffusion</subject><subject>Electrophoresis</subject><subject>Enrichment</subject><subject>Food Science</subject><subject>Gel electrophoresis</subject><subject>ICP-MS</subject><subject>Inductively coupled plasma mass spectrometry</subject><subject>Isotope dilution mass spectrometry</subject><subject>Isotope dilution techniques</subject><subject>Isotope Labeling</subject><subject>Isotopes</subject><subject>Isotopes - analysis</subject><subject>Labelling</subject><subject>Laboratory Medicine</subject><subject>Laser ablation</subject><subject>Liver</subject><subject>Liver - chemistry</subject><subject>Liver - enzymology</subject><subject>Mass Spectrometry</subject><subject>Metalloenzymes</subject><subject>Methods</subject><subject>Monitoring/Environmental Analysis</subject><subject>Original Paper</subject><subject>Physiological aspects</subject><subject>Properties</subject><subject>Protein quantification</subject><subject>Proteins</subject><subject>Proteomics</subject><subject>Separation</subject><subject>Sod</subject><subject>Speciation analysis</subject><subject>Species Specificity</subject><subject>Spectra</subject><subject>Spikes</subject><subject>Superoxide dismutase</subject><subject>Superoxide Dismutase - chemistry</subject><subject>Zinc</subject><subject>Zinc Isotopes - analysis</subject><issn>1618-2642</issn><issn>1618-2650</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkktr3TAQhU1padK0P6Cb1tBF04XTkWQ9vLykTwhkkWYtZHl0UbCtG48Nzb-vXKeBQknRYoTmO2cEc4riNYMzBqA_EgBnqgIGlVAGKvakOGaKmYorCU8f7jU_Kl4Q3QAwaZh6XhxxEI0wNRwXYddS6pcZy9vFjXMM0bs5prFMoaTlgFP6GTssu0jDMjvC8vTq8tOHcqE47ks6oI9I1e-alWWkNKfDimfL1cWNrr-jSC-LZ8H1hK_u60lx_eXzj_Nv1cXl1-_nu4vKSyXnymOras0Mtr6rjTbeu65DJZwXBgAlY9gxIT0P3gvfascht4NvVDC-5UKcFO8338OUbhek2Q6RPPa9GzEtZI1qpBFNXf-X1HUD3EipMnn6KMl0XWuQNdcZfbehe9ejjWNI8-T8itudaAxrBKh19Nk_qHw6HKJPI4aY3_8SsE3gp0Q0YbCHKQ5uurMM7JoEuyXB5iTYNQmWZc2b-18v7YDdg-LP6jPAN4Bya9zjZG_SMuV90aOubzdRcMm6_RTJXl9xYAKY0UJJLX4BfDrHiQ</recordid><startdate>20100801</startdate><enddate>20100801</enddate><creator>Deitrich, Christian L</creator><creator>Braukmann, Sandra</creator><creator>Raab, Andrea</creator><creator>Munro, Caroline</creator><creator>Pioselli, Barbara</creator><creator>Krupp, Eva M</creator><creator>Thomas-Oates, Jane E</creator><creator>Feldmann, Jörg</creator><general>Berlin/Heidelberg : Springer-Verlag</general><general>Springer-Verlag</general><general>Springer</general><scope>FBQ</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>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>L7M</scope><scope>7X8</scope><scope>7QH</scope><scope>7UA</scope><scope>C1K</scope></search><sort><creationdate>20100801</creationdate><title>Absolute quantification of superoxide dismutase (SOD) using species-specific isotope dilution analysis</title><author>Deitrich, Christian L ; Braukmann, Sandra ; Raab, Andrea ; Munro, Caroline ; Pioselli, Barbara ; Krupp, Eva M ; Thomas-Oates, Jane E ; Feldmann, Jörg</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c565t-ceb64718ebcd4878ccadde63ac3800e511ed135c2fcc3cb7a20de6fc96f8cb233</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Affinity labeling</topic><topic>Analysis</topic><topic>Analytical Chemistry</topic><topic>Animals</topic><topic>Biochemistry</topic><topic>Cattle</topic><topic>Characterization and Evaluation of Materials</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Chemistry Techniques, Analytical - methods</topic><topic>Copper - analysis</topic><topic>Diffusion</topic><topic>Electrophoresis</topic><topic>Enrichment</topic><topic>Food Science</topic><topic>Gel electrophoresis</topic><topic>ICP-MS</topic><topic>Inductively coupled plasma mass spectrometry</topic><topic>Isotope dilution mass spectrometry</topic><topic>Isotope dilution techniques</topic><topic>Isotope Labeling</topic><topic>Isotopes</topic><topic>Isotopes - analysis</topic><topic>Labelling</topic><topic>Laboratory Medicine</topic><topic>Laser ablation</topic><topic>Liver</topic><topic>Liver - chemistry</topic><topic>Liver - enzymology</topic><topic>Mass Spectrometry</topic><topic>Metalloenzymes</topic><topic>Methods</topic><topic>Monitoring/Environmental Analysis</topic><topic>Original Paper</topic><topic>Physiological aspects</topic><topic>Properties</topic><topic>Protein quantification</topic><topic>Proteins</topic><topic>Proteomics</topic><topic>Separation</topic><topic>Sod</topic><topic>Speciation analysis</topic><topic>Species Specificity</topic><topic>Spectra</topic><topic>Spikes</topic><topic>Superoxide dismutase</topic><topic>Superoxide Dismutase - chemistry</topic><topic>Zinc</topic><topic>Zinc Isotopes - analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Deitrich, Christian L</creatorcontrib><creatorcontrib>Braukmann, Sandra</creatorcontrib><creatorcontrib>Raab, Andrea</creatorcontrib><creatorcontrib>Munro, Caroline</creatorcontrib><creatorcontrib>Pioselli, Barbara</creatorcontrib><creatorcontrib>Krupp, Eva M</creatorcontrib><creatorcontrib>Thomas-Oates, Jane E</creatorcontrib><creatorcontrib>Feldmann, Jörg</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>MEDLINE - Academic</collection><collection>Aqualine</collection><collection>Water Resources Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Analytical and bioanalytical chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Deitrich, Christian L</au><au>Braukmann, Sandra</au><au>Raab, Andrea</au><au>Munro, Caroline</au><au>Pioselli, Barbara</au><au>Krupp, Eva M</au><au>Thomas-Oates, Jane E</au><au>Feldmann, Jörg</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Absolute quantification of superoxide dismutase (SOD) using species-specific isotope dilution analysis</atitle><jtitle>Analytical and bioanalytical chemistry</jtitle><stitle>Anal Bioanal Chem</stitle><addtitle>Anal Bioanal Chem</addtitle><date>2010-08-01</date><risdate>2010</risdate><volume>397</volume><issue>8</issue><spage>3515</spage><epage>3524</epage><pages>3515-3524</pages><issn>1618-2642</issn><eissn>1618-2650</eissn><abstract>Here we report for the first time the use of species-specific isotope dilution mass spectrometry for the absolute quantification of a metalloprotein using non-denaturing gel electrophoresis laser ablation inductively coupled plasma mass spectrometry (GE-LA-ICP-MS). The concept utilises the intrinsic metals of the metalloprotein for labelling of the isotopically labelled spike (⁶⁵Cu, ⁶⁸Zn SOD). The stability of the metal-protein complex under non-denaturing conditions during 1-D PAGE was confirmed and the performance of the method evaluated. Between 4 and 64 μg, SOD was quantified with a recovery rate between 82% and 110% in a standard. The use of the isotopically enriched SOD was utilised to identify the extent of orthogonal diffusion in 1-D gel electrophoresis. Orthogonal diffusion of natural and isotopically enriched SOD in the gel can interfere with the correct determination of the isotope ratios. The matrix effect of a cytosolic liver extract on the non-covalently bound copper and zinc in SOD was evaluated and no significant metal loss from the SOD spike was observed. This study represents the first step necessary for establishing and evaluating the use of a species-specific isotope dilution approach for the absolute quantification of SOD in real samples based on the combination of gel electrophoresis and LA-ICP-MS.</abstract><cop>Berlin/Heidelberg</cop><pub>Berlin/Heidelberg : Springer-Verlag</pub><pmid>20393840</pmid><doi>10.1007/s00216-010-3680-1</doi><tpages>10</tpages></addata></record> |
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| subjects | Affinity labeling Analysis Analytical Chemistry Animals Biochemistry Cattle Characterization and Evaluation of Materials Chemistry Chemistry and Materials Science Chemistry Techniques, Analytical - methods Copper - analysis Diffusion Electrophoresis Enrichment Food Science Gel electrophoresis ICP-MS Inductively coupled plasma mass spectrometry Isotope dilution mass spectrometry Isotope dilution techniques Isotope Labeling Isotopes Isotopes - analysis Labelling Laboratory Medicine Laser ablation Liver Liver - chemistry Liver - enzymology Mass Spectrometry Metalloenzymes Methods Monitoring/Environmental Analysis Original Paper Physiological aspects Properties Protein quantification Proteins Proteomics Separation Sod Speciation analysis Species Specificity Spectra Spikes Superoxide dismutase Superoxide Dismutase - chemistry Zinc Zinc Isotopes - analysis |
| title | Absolute quantification of superoxide dismutase (SOD) using species-specific isotope dilution analysis |
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