Reversed-phase HPLC of peptides: Assessing column and solvent selectivity on standard, polar-embedded and polar endcapped columns
We desired to evaluate the chromatographic selectivity for peptides of silica‐based RP high‐performance liquid chromatography stationary phases with various modifications (polar embedding and polar endcapping on C18 columns; ether‐linked phenyl column with polar endcapping) compared with n‐alkyl (C1...
Gespeichert in:
| Veröffentlicht in: | Journal of separation science 2010-10, Vol.33 (19), p.3005-3021 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 3021 |
|---|---|
| container_issue | 19 |
| container_start_page | 3005 |
| container_title | Journal of separation science |
| container_volume | 33 |
| creator | Mant, Colin T. Cepeniene, Dziuleta Hodges, Robert S. |
| description | We desired to evaluate the chromatographic selectivity for peptides of silica‐based RP high‐performance liquid chromatography stationary phases with various modifications (polar embedding and polar endcapping on C18 columns; ether‐linked phenyl column with polar endcapping) compared with n‐alkyl (C18, C8) and aromatic phenylhexyl columns. Thus, we have designed and synthesized two series of synthetic peptide standards with the sequence Gly–Gly–Leu–Gly–Gly–Ala–Leu–Gly–X–Leu–Lys–Lys–amide, where the N‐terminal either contains a free α‐amino group (AmC series) or is Nα‐acetylated (AcC series) and where position X is substituted by Gly, Ala, Val, Ile, Phe or Tyr. These represent series of peptides with single substitutions of n‐alkyl (Gly |
| doi_str_mv | 10.1002/jssc.201000518 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_762030709</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>762030709</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4488-b603997cea11a738d5c22e730d69a04e81ffd21fcf4a6c8a7cc20805750989f93</originalsourceid><addsrcrecordid>eNqFkE1v1DAQhi1ERUvhyhH5xoVsx3ES29zaFd1CV4BoEUfLa08gJV94stvukX9OtmlX3DjNaOZ5n8PL2CsBMwGQntwQ-VkK4w650E_YkShEnhgpsqf7HYpD9pzoBkAobeAZO0wFSJ3l-oj9-YobjIQh6X86Qn7xZTnnXcl77IcqIL3jp0RIVLU_uO_qddNy1wZOXb3BduCENfqh2lTDlnctp2F8uhje8r6rXUywWWEIGO4z9yeObfCu78fbpKMX7KB0NeHLh3nMvp2_v55fJMvPiw_z02Xis0zrZFWANEZ5dEI4JXXIfZqikhAK4yBDLcoypKL0ZeYKr53yPgUNucrBaFMaeczeTN4-dr_XSINtKvJY167Fbk1WFSlIULAjZxPpY0cUsbR9rBoXt1aA3bVud63bfetj4PWDer1qMOzxx5pHwEzAbVXj9j86-_Hqav6vPJmyFQ14t8-6-MsWSqrcfv-0sPni3MClOLPX8i8NbZ8_</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>762030709</pqid></control><display><type>article</type><title>Reversed-phase HPLC of peptides: Assessing column and solvent selectivity on standard, polar-embedded and polar endcapped columns</title><source>MEDLINE</source><source>Wiley Online Library Journals Frontfile Complete</source><creator>Mant, Colin T. ; Cepeniene, Dziuleta ; Hodges, Robert S.</creator><creatorcontrib>Mant, Colin T. ; Cepeniene, Dziuleta ; Hodges, Robert S.</creatorcontrib><description>We desired to evaluate the chromatographic selectivity for peptides of silica‐based RP high‐performance liquid chromatography stationary phases with various modifications (polar embedding and polar endcapping on C18 columns; ether‐linked phenyl column with polar endcapping) compared with n‐alkyl (C18, C8) and aromatic phenylhexyl columns. Thus, we have designed and synthesized two series of synthetic peptide standards with the sequence Gly–Gly–Leu–Gly–Gly–Ala–Leu–Gly–X–Leu–Lys–Lys–amide, where the N‐terminal either contains a free α‐amino group (AmC series) or is Nα‐acetylated (AcC series) and where position X is substituted by Gly, Ala, Val, Ile, Phe or Tyr. These represent series of peptides with single substitutions of n‐alkyl (Gly<Ala<Val<Ile in order of increasing hydrophobicity), alkyl versus aromatic side‐chains (Phe, Tyr) and a side‐chain also with polar character (Tyr). Peptide mixtures were run on all columns at pH 2 (linear gradients of ACN or methanol in aqueous mobile phases). Our initial results clearly demonstrate the useful complementarity of different RP high‐performance liquid chromatography packing materials, with peptide mixtures showing profound selectivity differences between n‐alkyl versus polar‐embedded and polar endcapped C18 stationary phases. In addition, changing the polarity of the RP matrix in combination with changing the polarity of the mobile phase (ACN versus methanol) is shown to be synergistic to maximize selectivity of peptide separations.</description><identifier>ISSN: 1615-9306</identifier><identifier>EISSN: 1615-9314</identifier><identifier>DOI: 10.1002/jssc.201000518</identifier><identifier>PMID: 21038458</identifier><language>eng</language><publisher>Weinheim: WILEY-VCH Verlag</publisher><subject>ACN ; Chromatography, High Pressure Liquid - methods ; Hydrophobic and Hydrophilic Interactions ; Methanol ; Methanol - chemistry ; Peptides ; Peptides - chemistry ; RP-HPLC ; Silicon Dioxide - chemistry ; Solvents - chemistry ; Stationary phases</subject><ispartof>Journal of separation science, 2010-10, Vol.33 (19), p.3005-3021</ispartof><rights>Copyright © 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4488-b603997cea11a738d5c22e730d69a04e81ffd21fcf4a6c8a7cc20805750989f93</citedby><cites>FETCH-LOGICAL-c4488-b603997cea11a738d5c22e730d69a04e81ffd21fcf4a6c8a7cc20805750989f93</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%2Fjssc.201000518$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjssc.201000518$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21038458$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mant, Colin T.</creatorcontrib><creatorcontrib>Cepeniene, Dziuleta</creatorcontrib><creatorcontrib>Hodges, Robert S.</creatorcontrib><title>Reversed-phase HPLC of peptides: Assessing column and solvent selectivity on standard, polar-embedded and polar endcapped columns</title><title>Journal of separation science</title><addtitle>J. Sep. Science</addtitle><description>We desired to evaluate the chromatographic selectivity for peptides of silica‐based RP high‐performance liquid chromatography stationary phases with various modifications (polar embedding and polar endcapping on C18 columns; ether‐linked phenyl column with polar endcapping) compared with n‐alkyl (C18, C8) and aromatic phenylhexyl columns. Thus, we have designed and synthesized two series of synthetic peptide standards with the sequence Gly–Gly–Leu–Gly–Gly–Ala–Leu–Gly–X–Leu–Lys–Lys–amide, where the N‐terminal either contains a free α‐amino group (AmC series) or is Nα‐acetylated (AcC series) and where position X is substituted by Gly, Ala, Val, Ile, Phe or Tyr. These represent series of peptides with single substitutions of n‐alkyl (Gly<Ala<Val<Ile in order of increasing hydrophobicity), alkyl versus aromatic side‐chains (Phe, Tyr) and a side‐chain also with polar character (Tyr). Peptide mixtures were run on all columns at pH 2 (linear gradients of ACN or methanol in aqueous mobile phases). Our initial results clearly demonstrate the useful complementarity of different RP high‐performance liquid chromatography packing materials, with peptide mixtures showing profound selectivity differences between n‐alkyl versus polar‐embedded and polar endcapped C18 stationary phases. In addition, changing the polarity of the RP matrix in combination with changing the polarity of the mobile phase (ACN versus methanol) is shown to be synergistic to maximize selectivity of peptide separations.</description><subject>ACN</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Hydrophobic and Hydrophilic Interactions</subject><subject>Methanol</subject><subject>Methanol - chemistry</subject><subject>Peptides</subject><subject>Peptides - chemistry</subject><subject>RP-HPLC</subject><subject>Silicon Dioxide - chemistry</subject><subject>Solvents - chemistry</subject><subject>Stationary phases</subject><issn>1615-9306</issn><issn>1615-9314</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1v1DAQhi1ERUvhyhH5xoVsx3ES29zaFd1CV4BoEUfLa08gJV94stvukX9OtmlX3DjNaOZ5n8PL2CsBMwGQntwQ-VkK4w650E_YkShEnhgpsqf7HYpD9pzoBkAobeAZO0wFSJ3l-oj9-YobjIQh6X86Qn7xZTnnXcl77IcqIL3jp0RIVLU_uO_qddNy1wZOXb3BduCENfqh2lTDlnctp2F8uhje8r6rXUywWWEIGO4z9yeObfCu78fbpKMX7KB0NeHLh3nMvp2_v55fJMvPiw_z02Xis0zrZFWANEZ5dEI4JXXIfZqikhAK4yBDLcoypKL0ZeYKr53yPgUNucrBaFMaeczeTN4-dr_XSINtKvJY167Fbk1WFSlIULAjZxPpY0cUsbR9rBoXt1aA3bVud63bfetj4PWDer1qMOzxx5pHwEzAbVXj9j86-_Hqav6vPJmyFQ14t8-6-MsWSqrcfv-0sPni3MClOLPX8i8NbZ8_</recordid><startdate>201010</startdate><enddate>201010</enddate><creator>Mant, Colin T.</creator><creator>Cepeniene, Dziuleta</creator><creator>Hodges, Robert S.</creator><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</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>201010</creationdate><title>Reversed-phase HPLC of peptides: Assessing column and solvent selectivity on standard, polar-embedded and polar endcapped columns</title><author>Mant, Colin T. ; Cepeniene, Dziuleta ; Hodges, Robert S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4488-b603997cea11a738d5c22e730d69a04e81ffd21fcf4a6c8a7cc20805750989f93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>ACN</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Hydrophobic and Hydrophilic Interactions</topic><topic>Methanol</topic><topic>Methanol - chemistry</topic><topic>Peptides</topic><topic>Peptides - chemistry</topic><topic>RP-HPLC</topic><topic>Silicon Dioxide - chemistry</topic><topic>Solvents - chemistry</topic><topic>Stationary phases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mant, Colin T.</creatorcontrib><creatorcontrib>Cepeniene, Dziuleta</creatorcontrib><creatorcontrib>Hodges, Robert S.</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>Journal of separation science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mant, Colin T.</au><au>Cepeniene, Dziuleta</au><au>Hodges, Robert S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Reversed-phase HPLC of peptides: Assessing column and solvent selectivity on standard, polar-embedded and polar endcapped columns</atitle><jtitle>Journal of separation science</jtitle><addtitle>J. Sep. Science</addtitle><date>2010-10</date><risdate>2010</risdate><volume>33</volume><issue>19</issue><spage>3005</spage><epage>3021</epage><pages>3005-3021</pages><issn>1615-9306</issn><eissn>1615-9314</eissn><abstract>We desired to evaluate the chromatographic selectivity for peptides of silica‐based RP high‐performance liquid chromatography stationary phases with various modifications (polar embedding and polar endcapping on C18 columns; ether‐linked phenyl column with polar endcapping) compared with n‐alkyl (C18, C8) and aromatic phenylhexyl columns. Thus, we have designed and synthesized two series of synthetic peptide standards with the sequence Gly–Gly–Leu–Gly–Gly–Ala–Leu–Gly–X–Leu–Lys–Lys–amide, where the N‐terminal either contains a free α‐amino group (AmC series) or is Nα‐acetylated (AcC series) and where position X is substituted by Gly, Ala, Val, Ile, Phe or Tyr. These represent series of peptides with single substitutions of n‐alkyl (Gly<Ala<Val<Ile in order of increasing hydrophobicity), alkyl versus aromatic side‐chains (Phe, Tyr) and a side‐chain also with polar character (Tyr). Peptide mixtures were run on all columns at pH 2 (linear gradients of ACN or methanol in aqueous mobile phases). Our initial results clearly demonstrate the useful complementarity of different RP high‐performance liquid chromatography packing materials, with peptide mixtures showing profound selectivity differences between n‐alkyl versus polar‐embedded and polar endcapped C18 stationary phases. In addition, changing the polarity of the RP matrix in combination with changing the polarity of the mobile phase (ACN versus methanol) is shown to be synergistic to maximize selectivity of peptide separations.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>21038458</pmid><doi>10.1002/jssc.201000518</doi><tpages>17</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1615-9306 |
| ispartof | Journal of separation science, 2010-10, Vol.33 (19), p.3005-3021 |
| issn | 1615-9306 1615-9314 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_762030709 |
| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | ACN Chromatography, High Pressure Liquid - methods Hydrophobic and Hydrophilic Interactions Methanol Methanol - chemistry Peptides Peptides - chemistry RP-HPLC Silicon Dioxide - chemistry Solvents - chemistry Stationary phases |
| title | Reversed-phase HPLC of peptides: Assessing column and solvent selectivity on standard, polar-embedded and polar endcapped columns |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-04T00%3A08%3A37IST&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=Reversed-phase%20HPLC%20of%20peptides:%20Assessing%20column%20and%20solvent%20selectivity%20on%20standard,%20polar-embedded%20and%20polar%20endcapped%20columns&rft.jtitle=Journal%20of%20separation%20science&rft.au=Mant,%20Colin%20T.&rft.date=2010-10&rft.volume=33&rft.issue=19&rft.spage=3005&rft.epage=3021&rft.pages=3005-3021&rft.issn=1615-9306&rft.eissn=1615-9314&rft_id=info:doi/10.1002/jssc.201000518&rft_dat=%3Cproquest_cross%3E762030709%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=762030709&rft_id=info:pmid/21038458&rfr_iscdi=true |