A novel high-throughput method for supported liquid extraction of retinol and alpha-tocopherol from human serum and simultaneous quantitation by liquid chromatography tandem mass spectrometry
Background Measurement of vitamin A (retinol) and E (alpha-tocopherol) in UK clinical laboratories is currently performed exclusively by high-performance liquid chromatography with ultraviolet detection. We investigated whether retinol and alpha-tocopherol could be measured simultaneously by liquid...
Gespeichert in:
| Veröffentlicht in: | Annals of clinical biochemistry 2016-07, Vol.53 (4), p.434-445 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 445 |
|---|---|
| container_issue | 4 |
| container_start_page | 434 |
| container_title | Annals of clinical biochemistry |
| container_volume | 53 |
| creator | Hinchliffe, Edward Rudge, James Reed, Paul |
| description | Background
Measurement of vitamin A (retinol) and E (alpha-tocopherol) in UK clinical laboratories is currently performed exclusively by high-performance liquid chromatography with ultraviolet detection. We investigated whether retinol and alpha-tocopherol could be measured simultaneously by liquid chromatography tandem mass spectrometry.
Methods
Serum samples (100 μL) were extracted using Isolute + Supported Liquid Extraction plates. Chromatography was performed on a Phenomenex Kinetex Biphenyl 2.6 μm, 50 × 2.1 mm column, and liquid chromatography tandem mass spectrometry on a Waters Acquity TQD. Injection-to-injection time was 4.3 min. The assay was validated according to published guidelines. Patient samples were used to compare liquid chromatography tandem mass spectrometry and high-performance liquid chromatography with ultraviolet detection methods.
Results
For retinol and alpha-tocopherol, respectively, the assay was linear up to 6.0 and 80.0 μmol/L, and lower limit of quantification was 0.07 and 0.26 μmol/L. Intra and interassay imprecision were within desirable analytical specifications. Analysis of quality control material aligned to NIST SRM 968e, and relative spiked recovery from human serum, both yielded results within 15% of target values. Method comparison with high-performance liquid chromatography with ultraviolet detection methodology demonstrated a negative bias for retinol and alpha-tocopherol by the liquid chromatography tandem mass spectrometry method. Analysis of United Kingdom National External Quality Assurance Scheme samples yielded mean bias from the target value of +3.0% for retinol and −11.2% for alpha-tocopherol.
Conclusions
We have developed a novel, high-throughput method for extraction of retinol and alpha-tocopherol from human serum followed by simultaneous quantitation by liquid chromatography tandem mass spectrometry. The method offers a rapid, sensitive, specific and cost-effective alternative to high-performance liquid chromatography with ultraviolet detection methodology, and is suitable for routine clinical monitoring of patients predisposed to fat-soluble vitamin malabsorption. |
| doi_str_mv | 10.1177/0004563215596024 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1793216920</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sage_id>10.1177_0004563215596024</sage_id><sourcerecordid>1793216920</sourcerecordid><originalsourceid>FETCH-LOGICAL-c379t-9c36ecfbc32b8eecacf7d5847b5b5a3778e7f13bc4e884c0625ee483ec774bc33</originalsourceid><addsrcrecordid>eNp1kc1u1DAUhS0EotPCnhW6SzYBO4njZFlVFJAqsYF15Dg3Y1dx7PEPYp6ur4an07JAYmXZ5zvn-uoQ8o7Rj4wJ8YlS2vKuqRnnQ0fr9gXZMcH7ilHKXpLdSa5O-gW5jPG-XGtB6WtyUXds4G0vduThGjb3C1fQZq-rpIPLe-1zAotJuxkWFyBm711IOMNqDtnMgL9TkCoZt4FbIGAym1tBbjPI1WtZJaec1xjK4xKcBZ2t3CBiyPaRisbmNckNXY5wyHJLJsnHuOn4PEOVr1iZ3D5Ir49Q6BktWBkjRI8qFRVTOL4hrxa5Rnz7dF6Rn7eff9x8re6-f_l2c31XqUYMqRpU06FaJtXUU4-opFrEzPtWTHzishGiR7GwZlIt9n2raFdzxLZvUAnRFldzRT6cc31wh4wxjdZEhet63mJkYig1dENNC0rPqAouxoDL6IOxMhxHRsdTbeO_tRXL-6f0PFmc_xqeeypAdQai3ON473LYyrb_D_wDKdemww</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1793216920</pqid></control><display><type>article</type><title>A novel high-throughput method for supported liquid extraction of retinol and alpha-tocopherol from human serum and simultaneous quantitation by liquid chromatography tandem mass spectrometry</title><source>MEDLINE</source><source>SAGE Complete A-Z List</source><creator>Hinchliffe, Edward ; Rudge, James ; Reed, Paul</creator><creatorcontrib>Hinchliffe, Edward ; Rudge, James ; Reed, Paul</creatorcontrib><description>Background
Measurement of vitamin A (retinol) and E (alpha-tocopherol) in UK clinical laboratories is currently performed exclusively by high-performance liquid chromatography with ultraviolet detection. We investigated whether retinol and alpha-tocopherol could be measured simultaneously by liquid chromatography tandem mass spectrometry.
Methods
Serum samples (100 μL) were extracted using Isolute + Supported Liquid Extraction plates. Chromatography was performed on a Phenomenex Kinetex Biphenyl 2.6 μm, 50 × 2.1 mm column, and liquid chromatography tandem mass spectrometry on a Waters Acquity TQD. Injection-to-injection time was 4.3 min. The assay was validated according to published guidelines. Patient samples were used to compare liquid chromatography tandem mass spectrometry and high-performance liquid chromatography with ultraviolet detection methods.
Results
For retinol and alpha-tocopherol, respectively, the assay was linear up to 6.0 and 80.0 μmol/L, and lower limit of quantification was 0.07 and 0.26 μmol/L. Intra and interassay imprecision were within desirable analytical specifications. Analysis of quality control material aligned to NIST SRM 968e, and relative spiked recovery from human serum, both yielded results within 15% of target values. Method comparison with high-performance liquid chromatography with ultraviolet detection methodology demonstrated a negative bias for retinol and alpha-tocopherol by the liquid chromatography tandem mass spectrometry method. Analysis of United Kingdom National External Quality Assurance Scheme samples yielded mean bias from the target value of +3.0% for retinol and −11.2% for alpha-tocopherol.
Conclusions
We have developed a novel, high-throughput method for extraction of retinol and alpha-tocopherol from human serum followed by simultaneous quantitation by liquid chromatography tandem mass spectrometry. The method offers a rapid, sensitive, specific and cost-effective alternative to high-performance liquid chromatography with ultraviolet detection methodology, and is suitable for routine clinical monitoring of patients predisposed to fat-soluble vitamin malabsorption.</description><identifier>ISSN: 0004-5632</identifier><identifier>EISSN: 1758-1001</identifier><identifier>DOI: 10.1177/0004563215596024</identifier><identifier>PMID: 26195487</identifier><language>eng</language><publisher>London, England: SAGE Publications</publisher><subject>Adult ; Aged ; alpha-Tocopherol - blood ; alpha-Tocopherol - isolation & purification ; Blood Chemical Analysis - methods ; Blood Chemical Analysis - standards ; Calibration ; Chromatography, High Pressure Liquid - standards ; Female ; Humans ; Male ; Middle Aged ; Reference Standards ; Reproducibility of Results ; Tandem Mass Spectrometry - standards ; Vitamin A - blood ; Vitamin A - isolation & purification</subject><ispartof>Annals of clinical biochemistry, 2016-07, Vol.53 (4), p.434-445</ispartof><rights>The Author(s) 2015</rights><rights>The Author(s) 2015.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c379t-9c36ecfbc32b8eecacf7d5847b5b5a3778e7f13bc4e884c0625ee483ec774bc33</citedby><cites>FETCH-LOGICAL-c379t-9c36ecfbc32b8eecacf7d5847b5b5a3778e7f13bc4e884c0625ee483ec774bc33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://journals.sagepub.com/doi/pdf/10.1177/0004563215596024$$EPDF$$P50$$Gsage$$H</linktopdf><linktohtml>$$Uhttps://journals.sagepub.com/doi/10.1177/0004563215596024$$EHTML$$P50$$Gsage$$H</linktohtml><link.rule.ids>314,776,780,21799,27903,27904,43600,43601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26195487$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hinchliffe, Edward</creatorcontrib><creatorcontrib>Rudge, James</creatorcontrib><creatorcontrib>Reed, Paul</creatorcontrib><title>A novel high-throughput method for supported liquid extraction of retinol and alpha-tocopherol from human serum and simultaneous quantitation by liquid chromatography tandem mass spectrometry</title><title>Annals of clinical biochemistry</title><addtitle>Ann Clin Biochem</addtitle><description>Background
Measurement of vitamin A (retinol) and E (alpha-tocopherol) in UK clinical laboratories is currently performed exclusively by high-performance liquid chromatography with ultraviolet detection. We investigated whether retinol and alpha-tocopherol could be measured simultaneously by liquid chromatography tandem mass spectrometry.
Methods
Serum samples (100 μL) were extracted using Isolute + Supported Liquid Extraction plates. Chromatography was performed on a Phenomenex Kinetex Biphenyl 2.6 μm, 50 × 2.1 mm column, and liquid chromatography tandem mass spectrometry on a Waters Acquity TQD. Injection-to-injection time was 4.3 min. The assay was validated according to published guidelines. Patient samples were used to compare liquid chromatography tandem mass spectrometry and high-performance liquid chromatography with ultraviolet detection methods.
Results
For retinol and alpha-tocopherol, respectively, the assay was linear up to 6.0 and 80.0 μmol/L, and lower limit of quantification was 0.07 and 0.26 μmol/L. Intra and interassay imprecision were within desirable analytical specifications. Analysis of quality control material aligned to NIST SRM 968e, and relative spiked recovery from human serum, both yielded results within 15% of target values. Method comparison with high-performance liquid chromatography with ultraviolet detection methodology demonstrated a negative bias for retinol and alpha-tocopherol by the liquid chromatography tandem mass spectrometry method. Analysis of United Kingdom National External Quality Assurance Scheme samples yielded mean bias from the target value of +3.0% for retinol and −11.2% for alpha-tocopherol.
Conclusions
We have developed a novel, high-throughput method for extraction of retinol and alpha-tocopherol from human serum followed by simultaneous quantitation by liquid chromatography tandem mass spectrometry. The method offers a rapid, sensitive, specific and cost-effective alternative to high-performance liquid chromatography with ultraviolet detection methodology, and is suitable for routine clinical monitoring of patients predisposed to fat-soluble vitamin malabsorption.</description><subject>Adult</subject><subject>Aged</subject><subject>alpha-Tocopherol - blood</subject><subject>alpha-Tocopherol - isolation & purification</subject><subject>Blood Chemical Analysis - methods</subject><subject>Blood Chemical Analysis - standards</subject><subject>Calibration</subject><subject>Chromatography, High Pressure Liquid - standards</subject><subject>Female</subject><subject>Humans</subject><subject>Male</subject><subject>Middle Aged</subject><subject>Reference Standards</subject><subject>Reproducibility of Results</subject><subject>Tandem Mass Spectrometry - standards</subject><subject>Vitamin A - blood</subject><subject>Vitamin A - isolation & purification</subject><issn>0004-5632</issn><issn>1758-1001</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kc1u1DAUhS0EotPCnhW6SzYBO4njZFlVFJAqsYF15Dg3Y1dx7PEPYp6ur4an07JAYmXZ5zvn-uoQ8o7Rj4wJ8YlS2vKuqRnnQ0fr9gXZMcH7ilHKXpLdSa5O-gW5jPG-XGtB6WtyUXds4G0vduThGjb3C1fQZq-rpIPLe-1zAotJuxkWFyBm711IOMNqDtnMgL9TkCoZt4FbIGAym1tBbjPI1WtZJaec1xjK4xKcBZ2t3CBiyPaRisbmNckNXY5wyHJLJsnHuOn4PEOVr1iZ3D5Ir49Q6BktWBkjRI8qFRVTOL4hrxa5Rnz7dF6Rn7eff9x8re6-f_l2c31XqUYMqRpU06FaJtXUU4-opFrEzPtWTHzishGiR7GwZlIt9n2raFdzxLZvUAnRFldzRT6cc31wh4wxjdZEhet63mJkYig1dENNC0rPqAouxoDL6IOxMhxHRsdTbeO_tRXL-6f0PFmc_xqeeypAdQai3ON473LYyrb_D_wDKdemww</recordid><startdate>20160701</startdate><enddate>20160701</enddate><creator>Hinchliffe, Edward</creator><creator>Rudge, James</creator><creator>Reed, Paul</creator><general>SAGE Publications</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>7X8</scope></search><sort><creationdate>20160701</creationdate><title>A novel high-throughput method for supported liquid extraction of retinol and alpha-tocopherol from human serum and simultaneous quantitation by liquid chromatography tandem mass spectrometry</title><author>Hinchliffe, Edward ; Rudge, James ; Reed, Paul</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c379t-9c36ecfbc32b8eecacf7d5847b5b5a3778e7f13bc4e884c0625ee483ec774bc33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Adult</topic><topic>Aged</topic><topic>alpha-Tocopherol - blood</topic><topic>alpha-Tocopherol - isolation & purification</topic><topic>Blood Chemical Analysis - methods</topic><topic>Blood Chemical Analysis - standards</topic><topic>Calibration</topic><topic>Chromatography, High Pressure Liquid - standards</topic><topic>Female</topic><topic>Humans</topic><topic>Male</topic><topic>Middle Aged</topic><topic>Reference Standards</topic><topic>Reproducibility of Results</topic><topic>Tandem Mass Spectrometry - standards</topic><topic>Vitamin A - blood</topic><topic>Vitamin A - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hinchliffe, Edward</creatorcontrib><creatorcontrib>Rudge, James</creatorcontrib><creatorcontrib>Reed, Paul</creatorcontrib><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>Annals of clinical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hinchliffe, Edward</au><au>Rudge, James</au><au>Reed, Paul</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A novel high-throughput method for supported liquid extraction of retinol and alpha-tocopherol from human serum and simultaneous quantitation by liquid chromatography tandem mass spectrometry</atitle><jtitle>Annals of clinical biochemistry</jtitle><addtitle>Ann Clin Biochem</addtitle><date>2016-07-01</date><risdate>2016</risdate><volume>53</volume><issue>4</issue><spage>434</spage><epage>445</epage><pages>434-445</pages><issn>0004-5632</issn><eissn>1758-1001</eissn><abstract>Background
Measurement of vitamin A (retinol) and E (alpha-tocopherol) in UK clinical laboratories is currently performed exclusively by high-performance liquid chromatography with ultraviolet detection. We investigated whether retinol and alpha-tocopherol could be measured simultaneously by liquid chromatography tandem mass spectrometry.
Methods
Serum samples (100 μL) were extracted using Isolute + Supported Liquid Extraction plates. Chromatography was performed on a Phenomenex Kinetex Biphenyl 2.6 μm, 50 × 2.1 mm column, and liquid chromatography tandem mass spectrometry on a Waters Acquity TQD. Injection-to-injection time was 4.3 min. The assay was validated according to published guidelines. Patient samples were used to compare liquid chromatography tandem mass spectrometry and high-performance liquid chromatography with ultraviolet detection methods.
Results
For retinol and alpha-tocopherol, respectively, the assay was linear up to 6.0 and 80.0 μmol/L, and lower limit of quantification was 0.07 and 0.26 μmol/L. Intra and interassay imprecision were within desirable analytical specifications. Analysis of quality control material aligned to NIST SRM 968e, and relative spiked recovery from human serum, both yielded results within 15% of target values. Method comparison with high-performance liquid chromatography with ultraviolet detection methodology demonstrated a negative bias for retinol and alpha-tocopherol by the liquid chromatography tandem mass spectrometry method. Analysis of United Kingdom National External Quality Assurance Scheme samples yielded mean bias from the target value of +3.0% for retinol and −11.2% for alpha-tocopherol.
Conclusions
We have developed a novel, high-throughput method for extraction of retinol and alpha-tocopherol from human serum followed by simultaneous quantitation by liquid chromatography tandem mass spectrometry. The method offers a rapid, sensitive, specific and cost-effective alternative to high-performance liquid chromatography with ultraviolet detection methodology, and is suitable for routine clinical monitoring of patients predisposed to fat-soluble vitamin malabsorption.</abstract><cop>London, England</cop><pub>SAGE Publications</pub><pmid>26195487</pmid><doi>10.1177/0004563215596024</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0004-5632 |
| ispartof | Annals of clinical biochemistry, 2016-07, Vol.53 (4), p.434-445 |
| issn | 0004-5632 1758-1001 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1793216920 |
| source | MEDLINE; SAGE Complete A-Z List |
| subjects | Adult Aged alpha-Tocopherol - blood alpha-Tocopherol - isolation & purification Blood Chemical Analysis - methods Blood Chemical Analysis - standards Calibration Chromatography, High Pressure Liquid - standards Female Humans Male Middle Aged Reference Standards Reproducibility of Results Tandem Mass Spectrometry - standards Vitamin A - blood Vitamin A - isolation & purification |
| title | A novel high-throughput method for supported liquid extraction of retinol and alpha-tocopherol from human serum and simultaneous quantitation by liquid chromatography tandem mass spectrometry |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-23T23%3A18%3A24IST&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=A%20novel%20high-throughput%20method%20for%20supported%20liquid%20extraction%20of%20retinol%20and%20alpha-tocopherol%20from%20human%20serum%20and%20simultaneous%20quantitation%20by%20liquid%20chromatography%20tandem%20mass%20spectrometry&rft.jtitle=Annals%20of%20clinical%20biochemistry&rft.au=Hinchliffe,%20Edward&rft.date=2016-07-01&rft.volume=53&rft.issue=4&rft.spage=434&rft.epage=445&rft.pages=434-445&rft.issn=0004-5632&rft.eissn=1758-1001&rft_id=info:doi/10.1177/0004563215596024&rft_dat=%3Cproquest_cross%3E1793216920%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=1793216920&rft_id=info:pmid/26195487&rft_sage_id=10.1177_0004563215596024&rfr_iscdi=true |