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
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
Background and aims: 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.
Materials and methods: 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.
Results: 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.
Conclusion: 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.
Bile acid, Branch chain alpha keto acid, Fatty acid, Liquid chromatography tandem mass spectrometry, Liquid-liquid extraction, Short chain fatty acid
Ng, Daniel Zhi Wei
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Lee, Sean Xian Yu
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Ooi, Delicia Shu Qin
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Ta, Le Duc Huy
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Yap, Gaik Chin
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Tay, Carina Jing Xuan
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Huang, Chiung-Hui
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Tham, Elizabeth Huiwen
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Loo, Evelyn Xiu Ling
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Shek, Lynette P.C.
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Goh, Anne
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Bever, Hugo P.S. Van
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Teoh, Oon Hoe
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Lee, Yung Seng
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Yap, Fabian
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Tan, Kok Hian
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Chong, Yap-Seng
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Chan, Shiao Yng
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Eriksson, Johan Gunnar
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Godfrey, Keith M.
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Lee, Bee Wah
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Chan, Eric Chun Yong
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26 June 2023
Ng, Daniel Zhi Wei
283dc7a7-e363-433e-aa72-cc9c195d14ec
Lee, Sean Xian Yu
7bcbe0d0-2c39-41c3-8166-ebcebef08657
Ooi, Delicia Shu Qin
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Ta, Le Duc Huy
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Yap, Gaik Chin
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Tay, Carina Jing Xuan
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Huang, Chiung-Hui
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Tham, Elizabeth Huiwen
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Loo, Evelyn Xiu Ling
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Shek, Lynette P.C.
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Goh, Anne
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Bever, Hugo P.S. Van
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Teoh, Oon Hoe
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Lee, Yung Seng
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Yap, Fabian
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Tan, Kok Hian
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Chong, Yap-Seng
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Chan, Shiao Yng
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Eriksson, Johan Gunnar
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Godfrey, Keith M.
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Lee, Bee Wah
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Chan, Eric Chun Yong
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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 and Chan, Eric Chun Yong
(2023)
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.
Clinica Chimica Acta, 547, [117449].
(doi:10.1016/j.cca.2023.117449).
Abstract
Background and aims: 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.
Materials and methods: 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.
Results: 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.
Conclusion: 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.
Text
Ng et al. CCA 2023 - Manuscript_clean version
- Accepted Manuscript
More information
Accepted/In Press date: 13 June 2023
e-pub ahead of print date: 17 June 2023
Published date: 26 June 2023
Additional Information:
Funding Information:
We would like to express our appreciation to Tan Bee Jen and Lee Soon Fatt for the technical expertise that they have provided for the operation of the LC-MS/MS.
Keywords:
Bile acid, Branch chain alpha keto acid, Fatty acid, Liquid chromatography tandem mass spectrometry, Liquid-liquid extraction, Short chain fatty acid
Identifiers
Local EPrints ID: 481737
URI: http://eprints.soton.ac.uk/id/eprint/481737
ISSN: 0009-8981
PURE UUID: 3bb8103e-7586-4846-8c53-f3551d68e327
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Date deposited: 07 Sep 2023 16:31
Last modified: 26 Jun 2024 04:01
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Contributors
Author:
Daniel Zhi Wei Ng
Author:
Sean Xian Yu Lee
Author:
Delicia Shu Qin Ooi
Author:
Le Duc Huy Ta
Author:
Gaik Chin Yap
Author:
Carina Jing Xuan Tay
Author:
Chiung-Hui Huang
Author:
Elizabeth Huiwen Tham
Author:
Evelyn Xiu Ling Loo
Author:
Lynette P.C. Shek
Author:
Anne Goh
Author:
Hugo P.S. Van Bever
Author:
Oon Hoe Teoh
Author:
Yung Seng Lee
Author:
Fabian Yap
Author:
Kok Hian Tan
Author:
Yap-Seng Chong
Author:
Shiao Yng Chan
Author:
Johan Gunnar Eriksson
Author:
Bee Wah Lee
Author:
Eric Chun Yong Chan
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