Sensitive LC-MS/MS method for the temporal profiling of bile acids, fatty acids and branched-chain alpha-keto acids in maternal plasma during pregnancy and cord blood plasma at delivery
[Display omitted] •An LC/MS-MS method quantifying gut microbial and dietary-derived metabolites.•Linearity, accuracy, precision, matrix effect, recovery and stability were validated.•Metabolites of interest were chromatographically separated and quantified.•LLE extracted metabolites with adequate se...
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| Veröffentlicht in: | Clinica chimica acta 2023-07, Vol.547, p.117449-117449, Article 117449 |
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| creator | Ng, Daniel Zhi Wei Lee, Sean Xian Yu Ooi, Delicia Shu Qin Ta, Le Duc Huy Yap, Gaik Chin Tay, Carina Jing Xuan Huang, Chiung-Hui Tham, Elizabeth Huiwen Loo, Evelyn Xiu Ling Shek, Lynette P.C. Goh, Anne Bever, Hugo P.S. Van Teoh, Oon Hoe Lee, Yung Seng Yap, Fabian Tan, Kok Hian Chong, Yap-Seng Chan, Shiao Yng Eriksson, Johan Gunnar Godfrey, Keith M. Lee, Bee Wah Chan, Eric Chun Yong |
| description | [Display omitted]
•An LC/MS-MS method quantifying gut microbial and dietary-derived metabolites.•Linearity, accuracy, precision, matrix effect, recovery and stability were validated.•Metabolites of interest were chromatographically separated and quantified.•LLE extracted metabolites with adequate sensitivity and consistent recovery.
There are significant changes to the maternal inflammatory profile across pregnancy. Recent studies suggest that perturbations in maternal gut microbial and dietary-derived plasma metabolites over the course of pregnancy mediate inflammation through a complex interplay of immunomodulatory effects. Despite this body of evidence, there is currently no analytical method that is suitable for the simultaneous profiling of these metabolites within human plasma.
We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the high-throughput analysis of these metabolites in human plasma without derivatization. Plasma samples were processed using liquid–liquid extraction method with varying proportions of methyl tert-butyl ether, methanol, and water in a 3:10:2.5 ratio to reduce matrix effects.
LC-MS/MS detection was sufficiently sensitive to quantify these gut microbial and dietary-derived metabolites at physiological concentrations and linear calibration curves with r2 > 0.99 were obtained. Recovery was consistent across concentration levels. Stability experiments confirmed that up to 160 samples could be analyzed within a single batch. The method was validated and applied to analyse maternal plasma during the first and third trimester and cord blood plasma of 5 mothers.
This study validated a straightforward and sensitive LC-MS/MS method for the simultaneous quantitation of gut microbial and dietary-derived metabolites in human plasma within 9 minutes without prior sample derivatization. |
| doi_str_mv | 10.1016/j.cca.2023.117449 |
| format | Article |
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•An LC/MS-MS method quantifying gut microbial and dietary-derived metabolites.•Linearity, accuracy, precision, matrix effect, recovery and stability were validated.•Metabolites of interest were chromatographically separated and quantified.•LLE extracted metabolites with adequate sensitivity and consistent recovery.
There are significant changes to the maternal inflammatory profile across pregnancy. Recent studies suggest that perturbations in maternal gut microbial and dietary-derived plasma metabolites over the course of pregnancy mediate inflammation through a complex interplay of immunomodulatory effects. Despite this body of evidence, there is currently no analytical method that is suitable for the simultaneous profiling of these metabolites within human plasma.
We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the high-throughput analysis of these metabolites in human plasma without derivatization. Plasma samples were processed using liquid–liquid extraction method with varying proportions of methyl tert-butyl ether, methanol, and water in a 3:10:2.5 ratio to reduce matrix effects.
LC-MS/MS detection was sufficiently sensitive to quantify these gut microbial and dietary-derived metabolites at physiological concentrations and linear calibration curves with r2 > 0.99 were obtained. Recovery was consistent across concentration levels. Stability experiments confirmed that up to 160 samples could be analyzed within a single batch. The method was validated and applied to analyse maternal plasma during the first and third trimester and cord blood plasma of 5 mothers.
This study validated a straightforward and sensitive LC-MS/MS method for the simultaneous quantitation of gut microbial and dietary-derived metabolites in human plasma within 9 minutes without prior sample derivatization.</description><identifier>ISSN: 0009-8981</identifier><identifier>EISSN: 1873-3492</identifier><identifier>DOI: 10.1016/j.cca.2023.117449</identifier><identifier>PMID: 37331549</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Bile acid ; Bile Acids and Salts ; Branch chain alpha keto acid ; Chromatography, High Pressure Liquid - methods ; Chromatography, Liquid - methods ; Fatty acid ; Fatty Acids ; Female ; Humans ; Keto Acids ; Liquid chromatography tandem mass spectrometry ; Liquid-liquid extraction ; Plasma ; Pregnancy ; Short chain fatty acid ; Tandem Mass Spectrometry - methods</subject><ispartof>Clinica chimica acta, 2023-07, Vol.547, p.117449-117449, Article 117449</ispartof><rights>2023 Elsevier B.V.</rights><rights>Copyright © 2023 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c353t-3cc28aa82f05412b324f616d87144ac0e085a5449aa15911b9be6f8b8b588e093</citedby><cites>FETCH-LOGICAL-c353t-3cc28aa82f05412b324f616d87144ac0e085a5449aa15911b9be6f8b8b588e093</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0009898123002516$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/37331549$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ng, Daniel Zhi Wei</creatorcontrib><creatorcontrib>Lee, Sean Xian Yu</creatorcontrib><creatorcontrib>Ooi, Delicia Shu Qin</creatorcontrib><creatorcontrib>Ta, Le Duc Huy</creatorcontrib><creatorcontrib>Yap, Gaik Chin</creatorcontrib><creatorcontrib>Tay, Carina Jing Xuan</creatorcontrib><creatorcontrib>Huang, Chiung-Hui</creatorcontrib><creatorcontrib>Tham, Elizabeth Huiwen</creatorcontrib><creatorcontrib>Loo, Evelyn Xiu Ling</creatorcontrib><creatorcontrib>Shek, Lynette P.C.</creatorcontrib><creatorcontrib>Goh, Anne</creatorcontrib><creatorcontrib>Bever, Hugo P.S. Van</creatorcontrib><creatorcontrib>Teoh, Oon Hoe</creatorcontrib><creatorcontrib>Lee, Yung Seng</creatorcontrib><creatorcontrib>Yap, Fabian</creatorcontrib><creatorcontrib>Tan, Kok Hian</creatorcontrib><creatorcontrib>Chong, Yap-Seng</creatorcontrib><creatorcontrib>Chan, Shiao Yng</creatorcontrib><creatorcontrib>Eriksson, Johan Gunnar</creatorcontrib><creatorcontrib>Godfrey, Keith M.</creatorcontrib><creatorcontrib>Lee, Bee Wah</creatorcontrib><creatorcontrib>Chan, Eric Chun Yong</creatorcontrib><title>Sensitive LC-MS/MS method for the temporal profiling of bile acids, fatty acids and branched-chain alpha-keto acids in maternal plasma during pregnancy and cord blood plasma at delivery</title><title>Clinica chimica acta</title><addtitle>Clin Chim Acta</addtitle><description>[Display omitted]
•An LC/MS-MS method quantifying gut microbial and dietary-derived metabolites.•Linearity, accuracy, precision, matrix effect, recovery and stability were validated.•Metabolites of interest were chromatographically separated and quantified.•LLE extracted metabolites with adequate sensitivity and consistent recovery.
There are significant changes to the maternal inflammatory profile across pregnancy. Recent studies suggest that perturbations in maternal gut microbial and dietary-derived plasma metabolites over the course of pregnancy mediate inflammation through a complex interplay of immunomodulatory effects. Despite this body of evidence, there is currently no analytical method that is suitable for the simultaneous profiling of these metabolites within human plasma.
We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the high-throughput analysis of these metabolites in human plasma without derivatization. Plasma samples were processed using liquid–liquid extraction method with varying proportions of methyl tert-butyl ether, methanol, and water in a 3:10:2.5 ratio to reduce matrix effects.
LC-MS/MS detection was sufficiently sensitive to quantify these gut microbial and dietary-derived metabolites at physiological concentrations and linear calibration curves with r2 > 0.99 were obtained. Recovery was consistent across concentration levels. Stability experiments confirmed that up to 160 samples could be analyzed within a single batch. The method was validated and applied to analyse maternal plasma during the first and third trimester and cord blood plasma of 5 mothers.
This study validated a straightforward and sensitive LC-MS/MS method for the simultaneous quantitation of gut microbial and dietary-derived metabolites in human plasma within 9 minutes without prior sample derivatization.</description><subject>Bile acid</subject><subject>Bile Acids and Salts</subject><subject>Branch chain alpha keto acid</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Chromatography, Liquid - methods</subject><subject>Fatty acid</subject><subject>Fatty Acids</subject><subject>Female</subject><subject>Humans</subject><subject>Keto Acids</subject><subject>Liquid chromatography tandem mass spectrometry</subject><subject>Liquid-liquid extraction</subject><subject>Plasma</subject><subject>Pregnancy</subject><subject>Short chain fatty acid</subject><subject>Tandem Mass Spectrometry - methods</subject><issn>0009-8981</issn><issn>1873-3492</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kcGO0zAQhi0EYsvCA3BBPnIgXTt2EkecULWwSF1xKJytiT3ZuCRxsd2V-mi8HS7pcuRkjfXNp5n5CXnL2ZozXt_s18bAumSlWHPeSNk-IyuuGlEI2ZbPyYox1haqVfyKvIpxn0vJav6SXIlGCF7JdkV-73COLrlHpNtNcb-7ud_RCdPgLe19oGlAmnA6-AAjPQTfu9HND9T3tHMjUjDOxg-0h5ROS0FhtrQLMJsBbWEGcDOF8TBA8ROTvzD5b4KEYT5LR4gTUHsMZ_Eh4MOcm09_PcaHLBt9HuaCQaIWxzxtOL0mL3oYI765vNfkx-fb75u7Yvvty9fNp21hRCVSIYwpFYAqe1ZJXnailH3Na6saLiUYhkxVUOXbAfCq5bxrO6x71amuUgpZK67J-8Wb1_91xJj05KLBcYQZ_THqUpVNXUulREb5gprgYwzY60NwE4ST5kyfE9N7nRPT58T0kljueXfRH7sJ7b-Op4gy8HEBMC_56DDoaBzOBq0LaJK23v1H_wfqbKge</recordid><startdate>20230701</startdate><enddate>20230701</enddate><creator>Ng, Daniel Zhi Wei</creator><creator>Lee, Sean Xian Yu</creator><creator>Ooi, Delicia Shu Qin</creator><creator>Ta, Le Duc Huy</creator><creator>Yap, Gaik Chin</creator><creator>Tay, Carina Jing Xuan</creator><creator>Huang, Chiung-Hui</creator><creator>Tham, Elizabeth Huiwen</creator><creator>Loo, Evelyn Xiu Ling</creator><creator>Shek, Lynette P.C.</creator><creator>Goh, Anne</creator><creator>Bever, Hugo P.S. Van</creator><creator>Teoh, Oon Hoe</creator><creator>Lee, Yung Seng</creator><creator>Yap, Fabian</creator><creator>Tan, Kok Hian</creator><creator>Chong, Yap-Seng</creator><creator>Chan, Shiao Yng</creator><creator>Eriksson, Johan Gunnar</creator><creator>Godfrey, Keith M.</creator><creator>Lee, Bee Wah</creator><creator>Chan, Eric Chun Yong</creator><general>Elsevier B.V</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>20230701</creationdate><title>Sensitive LC-MS/MS method for the temporal profiling of bile acids, fatty acids and branched-chain alpha-keto acids in maternal plasma during pregnancy and cord blood plasma at delivery</title><author>Ng, Daniel Zhi Wei ; Lee, Sean Xian Yu ; Ooi, Delicia Shu Qin ; Ta, Le Duc Huy ; Yap, Gaik Chin ; Tay, Carina Jing Xuan ; Huang, Chiung-Hui ; Tham, Elizabeth Huiwen ; Loo, Evelyn Xiu Ling ; Shek, Lynette P.C. ; Goh, Anne ; Bever, Hugo P.S. Van ; Teoh, Oon Hoe ; Lee, Yung Seng ; Yap, Fabian ; Tan, Kok Hian ; Chong, Yap-Seng ; Chan, Shiao Yng ; Eriksson, Johan Gunnar ; Godfrey, Keith M. ; Lee, Bee Wah ; Chan, Eric Chun Yong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c353t-3cc28aa82f05412b324f616d87144ac0e085a5449aa15911b9be6f8b8b588e093</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Bile acid</topic><topic>Bile Acids and Salts</topic><topic>Branch chain alpha keto acid</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Chromatography, Liquid - methods</topic><topic>Fatty acid</topic><topic>Fatty Acids</topic><topic>Female</topic><topic>Humans</topic><topic>Keto Acids</topic><topic>Liquid chromatography tandem mass spectrometry</topic><topic>Liquid-liquid extraction</topic><topic>Plasma</topic><topic>Pregnancy</topic><topic>Short chain fatty acid</topic><topic>Tandem Mass Spectrometry - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ng, Daniel Zhi Wei</creatorcontrib><creatorcontrib>Lee, Sean Xian Yu</creatorcontrib><creatorcontrib>Ooi, Delicia Shu Qin</creatorcontrib><creatorcontrib>Ta, Le Duc Huy</creatorcontrib><creatorcontrib>Yap, Gaik Chin</creatorcontrib><creatorcontrib>Tay, Carina Jing Xuan</creatorcontrib><creatorcontrib>Huang, Chiung-Hui</creatorcontrib><creatorcontrib>Tham, Elizabeth Huiwen</creatorcontrib><creatorcontrib>Loo, Evelyn Xiu Ling</creatorcontrib><creatorcontrib>Shek, Lynette P.C.</creatorcontrib><creatorcontrib>Goh, Anne</creatorcontrib><creatorcontrib>Bever, Hugo P.S. Van</creatorcontrib><creatorcontrib>Teoh, Oon Hoe</creatorcontrib><creatorcontrib>Lee, Yung Seng</creatorcontrib><creatorcontrib>Yap, Fabian</creatorcontrib><creatorcontrib>Tan, Kok Hian</creatorcontrib><creatorcontrib>Chong, Yap-Seng</creatorcontrib><creatorcontrib>Chan, Shiao Yng</creatorcontrib><creatorcontrib>Eriksson, Johan Gunnar</creatorcontrib><creatorcontrib>Godfrey, Keith M.</creatorcontrib><creatorcontrib>Lee, Bee Wah</creatorcontrib><creatorcontrib>Chan, Eric Chun Yong</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>Clinica chimica acta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ng, Daniel Zhi Wei</au><au>Lee, Sean Xian Yu</au><au>Ooi, Delicia Shu Qin</au><au>Ta, Le Duc Huy</au><au>Yap, Gaik Chin</au><au>Tay, Carina Jing Xuan</au><au>Huang, Chiung-Hui</au><au>Tham, Elizabeth Huiwen</au><au>Loo, Evelyn Xiu Ling</au><au>Shek, Lynette P.C.</au><au>Goh, Anne</au><au>Bever, Hugo P.S. Van</au><au>Teoh, Oon Hoe</au><au>Lee, Yung Seng</au><au>Yap, Fabian</au><au>Tan, Kok Hian</au><au>Chong, Yap-Seng</au><au>Chan, Shiao Yng</au><au>Eriksson, Johan Gunnar</au><au>Godfrey, Keith M.</au><au>Lee, Bee Wah</au><au>Chan, Eric Chun Yong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sensitive LC-MS/MS method for the temporal profiling of bile acids, fatty acids and branched-chain alpha-keto acids in maternal plasma during pregnancy and cord blood plasma at delivery</atitle><jtitle>Clinica chimica acta</jtitle><addtitle>Clin Chim Acta</addtitle><date>2023-07-01</date><risdate>2023</risdate><volume>547</volume><spage>117449</spage><epage>117449</epage><pages>117449-117449</pages><artnum>117449</artnum><issn>0009-8981</issn><eissn>1873-3492</eissn><abstract>[Display omitted]
•An LC/MS-MS method quantifying gut microbial and dietary-derived metabolites.•Linearity, accuracy, precision, matrix effect, recovery and stability were validated.•Metabolites of interest were chromatographically separated and quantified.•LLE extracted metabolites with adequate sensitivity and consistent recovery.
There are significant changes to the maternal inflammatory profile across pregnancy. Recent studies suggest that perturbations in maternal gut microbial and dietary-derived plasma metabolites over the course of pregnancy mediate inflammation through a complex interplay of immunomodulatory effects. Despite this body of evidence, there is currently no analytical method that is suitable for the simultaneous profiling of these metabolites within human plasma.
We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the high-throughput analysis of these metabolites in human plasma without derivatization. Plasma samples were processed using liquid–liquid extraction method with varying proportions of methyl tert-butyl ether, methanol, and water in a 3:10:2.5 ratio to reduce matrix effects.
LC-MS/MS detection was sufficiently sensitive to quantify these gut microbial and dietary-derived metabolites at physiological concentrations and linear calibration curves with r2 > 0.99 were obtained. Recovery was consistent across concentration levels. Stability experiments confirmed that up to 160 samples could be analyzed within a single batch. The method was validated and applied to analyse maternal plasma during the first and third trimester and cord blood plasma of 5 mothers.
This study validated a straightforward and sensitive LC-MS/MS method for the simultaneous quantitation of gut microbial and dietary-derived metabolites in human plasma within 9 minutes without prior sample derivatization.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>37331549</pmid><doi>10.1016/j.cca.2023.117449</doi><tpages>1</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0009-8981 |
| ispartof | Clinica chimica acta, 2023-07, Vol.547, p.117449-117449, Article 117449 |
| issn | 0009-8981 1873-3492 |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Bile acid Bile Acids and Salts Branch chain alpha keto acid Chromatography, High Pressure Liquid - methods Chromatography, Liquid - methods Fatty acid Fatty Acids Female Humans Keto Acids Liquid chromatography tandem mass spectrometry Liquid-liquid extraction Plasma Pregnancy Short chain fatty acid Tandem Mass Spectrometry - methods |
| title | Sensitive LC-MS/MS method for the temporal profiling of bile acids, fatty acids and branched-chain alpha-keto acids in maternal plasma during pregnancy and cord blood plasma at delivery |
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