Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds
The l -tryptophan–derived tricyclic hydroperoxide cis -WOOH was recently identified as a novel and biologically important factor for regulating vascular tone and blood pressure under inflammatory conditions and potentially other cellular redox signaling events. cis -WOOH is highly labile and current...
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| Veröffentlicht in: | Nature protocols 2021-07, Vol.16 (7), p.3382-3418 |
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| creator | Queiroz, Raphael F. Suarna, Cacang Corcilius, Leo Sergeant, Genevieve E. Shengule, Sudhir Payne, Richard J. Ayer, Anita Stocker, Roland |
| description | The
l
-tryptophan–derived tricyclic hydroperoxide
cis
-WOOH was recently identified as a novel and biologically important factor for regulating vascular tone and blood pressure under inflammatory conditions and potentially other cellular redox signaling events.
cis
-WOOH is highly labile and currently not available commercially. In this protocol, we provide procedures for the synthesis, purification, quantification and characterization of
cis
-WOOH, its epimer
trans-
WOOH and their respective alcohols (
cis
-WOH and
trans
-WOH). Photo-oxidation of
l
-tryptophan (
l
-Trp) results in a mixture containing
cis
-WOOH and
trans
-WOOH, which are separated and purified by semi-preparative HPLC.
cis
-WOH and
trans
-WOH are then produced by sodium borohydride reduction and purified by semi-preparative HPLC. Characterization of
cis
-WOOH and
trans
-WOOH and the reduced alcohol variants is achieved using HPLC, fluorescence, NMR and liquid chromatography-tandem mass spectrometry. The protocol provides instructions for storage and quantification, as well as ways to test the stability of these hydroperoxides in commonly used buffers and media. Finally, we describe examples of how to monitor the formation of
cis
-WOOH in biological samples. The protocol ensures reasonable yield (11%) and purity (>99%) of
cis
-WOOH and control compounds in 5–6 d and outlines conditions under which
cis
-WOOH is stable for several months.
The hydroperoxide
l
-tryptophan derivative
cis
-WOOH regulates vascular tone under inflammatory conditions via redox signaling. This protocol describes the preparation of
cis
-WOOH and related compounds for use as analytical standards and biochemical tools. |
| doi_str_mv | 10.1038/s41596-021-00541-1 |
| format | Article |
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l
-tryptophan–derived tricyclic hydroperoxide
cis
-WOOH was recently identified as a novel and biologically important factor for regulating vascular tone and blood pressure under inflammatory conditions and potentially other cellular redox signaling events.
cis
-WOOH is highly labile and currently not available commercially. In this protocol, we provide procedures for the synthesis, purification, quantification and characterization of
cis
-WOOH, its epimer
trans-
WOOH and their respective alcohols (
cis
-WOH and
trans
-WOH). Photo-oxidation of
l
-tryptophan (
l
-Trp) results in a mixture containing
cis
-WOOH and
trans
-WOOH, which are separated and purified by semi-preparative HPLC.
cis
-WOH and
trans
-WOH are then produced by sodium borohydride reduction and purified by semi-preparative HPLC. Characterization of
cis
-WOOH and
trans
-WOOH and the reduced alcohol variants is achieved using HPLC, fluorescence, NMR and liquid chromatography-tandem mass spectrometry. The protocol provides instructions for storage and quantification, as well as ways to test the stability of these hydroperoxides in commonly used buffers and media. Finally, we describe examples of how to monitor the formation of
cis
-WOOH in biological samples. The protocol ensures reasonable yield (11%) and purity (>99%) of
cis
-WOOH and control compounds in 5–6 d and outlines conditions under which
cis
-WOOH is stable for several months.
The hydroperoxide
l
-tryptophan derivative
cis
-WOOH regulates vascular tone under inflammatory conditions via redox signaling. This protocol describes the preparation of
cis
-WOOH and related compounds for use as analytical standards and biochemical tools.</description><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/s41596-021-00541-1</identifier><identifier>PMID: 34117477</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>631/45/2783 ; 631/92/1643 ; Alcohol ; Alcohols ; Analytical Chemistry ; Animals ; Biological properties ; Biological samples ; Biological Techniques ; Biomedical and Life Sciences ; Blood pressure ; Cardiovascular agents ; Chemical properties ; Chemical research ; Chemical synthesis ; Chromatography ; Chromatography, High Pressure Liquid ; Clinical chemistry ; Computational Biology/Bioinformatics ; Fluorescence ; High performance liquid chromatography ; Humans ; Hydrogen Peroxide - chemical synthesis ; Hydrogen Peroxide - isolation & purification ; Inflammation ; Life Sciences ; Liquid chromatography ; Mass Spectrometry ; Mass spectroscopy ; Methods ; Microarrays ; NMR ; Nuclear magnetic resonance ; Organic Chemistry ; Oxidation ; Oxidation-Reduction ; Photooxidation ; Physiology ; Production processes ; Proteins ; Protocol ; Reproducibility of Results ; Scientific imaging ; Signal transduction ; Signaling ; Superoxides - chemistry ; Temperature ; Tryptophan ; Tryptophan - chemistry ; Vasoactive agents</subject><ispartof>Nature protocols, 2021-07, Vol.16 (7), p.3382-3418</ispartof><rights>The Author(s), under exclusive licence to Springer Nature Limited 2021</rights><rights>COPYRIGHT 2021 Nature Publishing Group</rights><rights>The Author(s), under exclusive licence to Springer Nature Limited 2021.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c476t-4ec130ba057293c514a3f6ecd6a7e168918a1f279ce7c05fe8b36b59efa605ea3</citedby><cites>FETCH-LOGICAL-c476t-4ec130ba057293c514a3f6ecd6a7e168918a1f279ce7c05fe8b36b59efa605ea3</cites><orcidid>0000-0002-3618-9226 ; 0000-0002-7757-4286 ; 0000-0003-2312-4922 ; 0000-0002-7073-2835 ; 0000-0002-7414-6681</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/s41596-021-00541-1$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/s41596-021-00541-1$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,777,781,27905,27906,41469,42538,51300</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34117477$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Queiroz, Raphael F.</creatorcontrib><creatorcontrib>Suarna, Cacang</creatorcontrib><creatorcontrib>Corcilius, Leo</creatorcontrib><creatorcontrib>Sergeant, Genevieve E.</creatorcontrib><creatorcontrib>Shengule, Sudhir</creatorcontrib><creatorcontrib>Payne, Richard J.</creatorcontrib><creatorcontrib>Ayer, Anita</creatorcontrib><creatorcontrib>Stocker, Roland</creatorcontrib><title>Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>The
l
-tryptophan–derived tricyclic hydroperoxide
cis
-WOOH was recently identified as a novel and biologically important factor for regulating vascular tone and blood pressure under inflammatory conditions and potentially other cellular redox signaling events.
cis
-WOOH is highly labile and currently not available commercially. In this protocol, we provide procedures for the synthesis, purification, quantification and characterization of
cis
-WOOH, its epimer
trans-
WOOH and their respective alcohols (
cis
-WOH and
trans
-WOH). Photo-oxidation of
l
-tryptophan (
l
-Trp) results in a mixture containing
cis
-WOOH and
trans
-WOOH, which are separated and purified by semi-preparative HPLC.
cis
-WOH and
trans
-WOH are then produced by sodium borohydride reduction and purified by semi-preparative HPLC. Characterization of
cis
-WOOH and
trans
-WOOH and the reduced alcohol variants is achieved using HPLC, fluorescence, NMR and liquid chromatography-tandem mass spectrometry. The protocol provides instructions for storage and quantification, as well as ways to test the stability of these hydroperoxides in commonly used buffers and media. Finally, we describe examples of how to monitor the formation of
cis
-WOOH in biological samples. The protocol ensures reasonable yield (11%) and purity (>99%) of
cis
-WOOH and control compounds in 5–6 d and outlines conditions under which
cis
-WOOH is stable for several months.
The hydroperoxide
l
-tryptophan derivative
cis
-WOOH regulates vascular tone under inflammatory conditions via redox signaling. This protocol describes the preparation of
cis
-WOOH and related compounds for use as analytical standards and biochemical tools.</description><subject>631/45/2783</subject><subject>631/92/1643</subject><subject>Alcohol</subject><subject>Alcohols</subject><subject>Analytical Chemistry</subject><subject>Animals</subject><subject>Biological properties</subject><subject>Biological samples</subject><subject>Biological Techniques</subject><subject>Biomedical and Life Sciences</subject><subject>Blood pressure</subject><subject>Cardiovascular agents</subject><subject>Chemical properties</subject><subject>Chemical research</subject><subject>Chemical synthesis</subject><subject>Chromatography</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Clinical chemistry</subject><subject>Computational Biology/Bioinformatics</subject><subject>Fluorescence</subject><subject>High performance liquid chromatography</subject><subject>Humans</subject><subject>Hydrogen Peroxide - chemical synthesis</subject><subject>Hydrogen Peroxide - isolation & purification</subject><subject>Inflammation</subject><subject>Life Sciences</subject><subject>Liquid chromatography</subject><subject>Mass Spectrometry</subject><subject>Mass spectroscopy</subject><subject>Methods</subject><subject>Microarrays</subject><subject>NMR</subject><subject>Nuclear magnetic resonance</subject><subject>Organic Chemistry</subject><subject>Oxidation</subject><subject>Oxidation-Reduction</subject><subject>Photooxidation</subject><subject>Physiology</subject><subject>Production processes</subject><subject>Proteins</subject><subject>Protocol</subject><subject>Reproducibility of Results</subject><subject>Scientific imaging</subject><subject>Signal transduction</subject><subject>Signaling</subject><subject>Superoxides - chemistry</subject><subject>Temperature</subject><subject>Tryptophan</subject><subject>Tryptophan - chemistry</subject><subject>Vasoactive agents</subject><issn>1754-2189</issn><issn>1750-2799</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kV1rFDEUhgdRbK3-AS9kwCvBqclMPmYuS6laKCh-XIezyck2ZSYZk8zS_fdmd6tlQSQXJ5w87wnveavqNSXnlHT9h8QoH0RDWtoQwhlt6JPqlEpOmlYOw9P9nTUt7YeT6kVKd4Qw2Qn5vDrpGKWSSXlaha8RZ4iQXfDv6w2MzuzvNXhTLwnrYGso_RRAZ7fBOsftnMN8C74xGEvH1LdbE8OMMdw7g3thxBE34HOtg88xjKVOc1i8SS-rZxbGhK8e6ln18-PVj8vPzc2XT9eXFzeNZlLkhqGmHVkB4bIdOs0pg84K1EaARCr6gfZAbbGpUWrCLfarTqz4gBYE4QjdWfX2MHeO4deCKau7sERfvlQtZwOTPen7R2oNIyrnbcgR9OSSVhdCyF6ystxCnf-DKsfg5IpDtK70jwTvjgS7LeB9XsOSkrr-_u2YbQ-sjiGliFbN0U0Qt4oStYtZHWJWBVb7mNVO9ObB3bKa0PyV_Mm1AN0BSOXJrzE-2v_P2N9RCLKx</recordid><startdate>20210701</startdate><enddate>20210701</enddate><creator>Queiroz, Raphael F.</creator><creator>Suarna, Cacang</creator><creator>Corcilius, Leo</creator><creator>Sergeant, Genevieve E.</creator><creator>Shengule, Sudhir</creator><creator>Payne, Richard J.</creator><creator>Ayer, Anita</creator><creator>Stocker, Roland</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><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>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PATMY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PYCSY</scope><scope>RC3</scope><orcidid>https://orcid.org/0000-0002-3618-9226</orcidid><orcidid>https://orcid.org/0000-0002-7757-4286</orcidid><orcidid>https://orcid.org/0000-0003-2312-4922</orcidid><orcidid>https://orcid.org/0000-0002-7073-2835</orcidid><orcidid>https://orcid.org/0000-0002-7414-6681</orcidid></search><sort><creationdate>20210701</creationdate><title>Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds</title><author>Queiroz, Raphael F. ; Suarna, Cacang ; Corcilius, Leo ; Sergeant, Genevieve E. ; Shengule, Sudhir ; Payne, Richard J. ; Ayer, Anita ; Stocker, Roland</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c476t-4ec130ba057293c514a3f6ecd6a7e168918a1f279ce7c05fe8b36b59efa605ea3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>631/45/2783</topic><topic>631/92/1643</topic><topic>Alcohol</topic><topic>Alcohols</topic><topic>Analytical Chemistry</topic><topic>Animals</topic><topic>Biological properties</topic><topic>Biological samples</topic><topic>Biological Techniques</topic><topic>Biomedical and Life Sciences</topic><topic>Blood pressure</topic><topic>Cardiovascular agents</topic><topic>Chemical properties</topic><topic>Chemical research</topic><topic>Chemical synthesis</topic><topic>Chromatography</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Clinical chemistry</topic><topic>Computational Biology/Bioinformatics</topic><topic>Fluorescence</topic><topic>High performance liquid chromatography</topic><topic>Humans</topic><topic>Hydrogen Peroxide - chemical synthesis</topic><topic>Hydrogen Peroxide - isolation & purification</topic><topic>Inflammation</topic><topic>Life Sciences</topic><topic>Liquid chromatography</topic><topic>Mass Spectrometry</topic><topic>Mass spectroscopy</topic><topic>Methods</topic><topic>Microarrays</topic><topic>NMR</topic><topic>Nuclear magnetic resonance</topic><topic>Organic Chemistry</topic><topic>Oxidation</topic><topic>Oxidation-Reduction</topic><topic>Photooxidation</topic><topic>Physiology</topic><topic>Production processes</topic><topic>Proteins</topic><topic>Protocol</topic><topic>Reproducibility of Results</topic><topic>Scientific imaging</topic><topic>Signal transduction</topic><topic>Signaling</topic><topic>Superoxides - chemistry</topic><topic>Temperature</topic><topic>Tryptophan</topic><topic>Tryptophan - chemistry</topic><topic>Vasoactive agents</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Queiroz, Raphael F.</creatorcontrib><creatorcontrib>Suarna, Cacang</creatorcontrib><creatorcontrib>Corcilius, Leo</creatorcontrib><creatorcontrib>Sergeant, Genevieve E.</creatorcontrib><creatorcontrib>Shengule, Sudhir</creatorcontrib><creatorcontrib>Payne, Richard J.</creatorcontrib><creatorcontrib>Ayer, Anita</creatorcontrib><creatorcontrib>Stocker, Roland</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Queiroz, Raphael F.</au><au>Suarna, Cacang</au><au>Corcilius, Leo</au><au>Sergeant, Genevieve E.</au><au>Shengule, Sudhir</au><au>Payne, Richard J.</au><au>Ayer, Anita</au><au>Stocker, Roland</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2021-07-01</date><risdate>2021</risdate><volume>16</volume><issue>7</issue><spage>3382</spage><epage>3418</epage><pages>3382-3418</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>The
l
-tryptophan–derived tricyclic hydroperoxide
cis
-WOOH was recently identified as a novel and biologically important factor for regulating vascular tone and blood pressure under inflammatory conditions and potentially other cellular redox signaling events.
cis
-WOOH is highly labile and currently not available commercially. In this protocol, we provide procedures for the synthesis, purification, quantification and characterization of
cis
-WOOH, its epimer
trans-
WOOH and their respective alcohols (
cis
-WOH and
trans
-WOH). Photo-oxidation of
l
-tryptophan (
l
-Trp) results in a mixture containing
cis
-WOOH and
trans
-WOOH, which are separated and purified by semi-preparative HPLC.
cis
-WOH and
trans
-WOH are then produced by sodium borohydride reduction and purified by semi-preparative HPLC. Characterization of
cis
-WOOH and
trans
-WOOH and the reduced alcohol variants is achieved using HPLC, fluorescence, NMR and liquid chromatography-tandem mass spectrometry. The protocol provides instructions for storage and quantification, as well as ways to test the stability of these hydroperoxides in commonly used buffers and media. Finally, we describe examples of how to monitor the formation of
cis
-WOOH in biological samples. The protocol ensures reasonable yield (11%) and purity (>99%) of
cis
-WOOH and control compounds in 5–6 d and outlines conditions under which
cis
-WOOH is stable for several months.
The hydroperoxide
l
-tryptophan derivative
cis
-WOOH regulates vascular tone under inflammatory conditions via redox signaling. This protocol describes the preparation of
cis
-WOOH and related compounds for use as analytical standards and biochemical tools.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>34117477</pmid><doi>10.1038/s41596-021-00541-1</doi><tpages>37</tpages><orcidid>https://orcid.org/0000-0002-3618-9226</orcidid><orcidid>https://orcid.org/0000-0002-7757-4286</orcidid><orcidid>https://orcid.org/0000-0003-2312-4922</orcidid><orcidid>https://orcid.org/0000-0002-7073-2835</orcidid><orcidid>https://orcid.org/0000-0002-7414-6681</orcidid></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1754-2189 |
| ispartof | Nature protocols, 2021-07, Vol.16 (7), p.3382-3418 |
| issn | 1754-2189 1750-2799 |
| language | eng |
| recordid | cdi_proquest_journals_2549478088 |
| source | MEDLINE; Springer Nature - Complete Springer Journals; Nature Journals Online |
| subjects | 631/45/2783 631/92/1643 Alcohol Alcohols Analytical Chemistry Animals Biological properties Biological samples Biological Techniques Biomedical and Life Sciences Blood pressure Cardiovascular agents Chemical properties Chemical research Chemical synthesis Chromatography Chromatography, High Pressure Liquid Clinical chemistry Computational Biology/Bioinformatics Fluorescence High performance liquid chromatography Humans Hydrogen Peroxide - chemical synthesis Hydrogen Peroxide - isolation & purification Inflammation Life Sciences Liquid chromatography Mass Spectrometry Mass spectroscopy Methods Microarrays NMR Nuclear magnetic resonance Organic Chemistry Oxidation Oxidation-Reduction Photooxidation Physiology Production processes Proteins Protocol Reproducibility of Results Scientific imaging Signal transduction Signaling Superoxides - chemistry Temperature Tryptophan Tryptophan - chemistry Vasoactive agents |
| title | Preparation, validation and use of a vasoactive tryptophan-derived hydroperoxide and relevant control compounds |
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