Structural elucidation and quantification of phenolic conjugates present in human urine after tea intake

J.J.J. van der Hooft, R.C.H. de Vos, V. Mihaleva, R.J. Bino, L.O. Ridder, N. de Roo, D.M. Jacobs, J.P.M. van Duynhoven, J.J.M. Vervoort

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Abstract

In dietary polyphenol exposure studies, annotation and identification of urinary metabolites present at low (micromolar) concentrations are major obstacles. In order to determine the biological activity of specific components, it is necessary to have the correct structures and the quantification of the polyphenol-derived conjugates present in the human body. We present a procedure for identification and quantification of metabolites and conjugates excreted in human urine after single bolus intake of black or green tea. A combination of a solid phase extraction (SPE) preparation step and two high pressure liquid chromatography (HPLC)-based analytical platforms was used; namely, accurate mass fragmentation (HPLC-FTMSn) and mass-guided SPE-trapping of selected compounds for nuclear magnetic resonance spectroscopy (NMR) measurements (HPLC-TOFMS-SPE-NMR). HPLC-FTMSn analysis led to the annotation of 138 urinary metabolites, including 48 valerolactone and valeric acid conjugates. By combining the results from MSn fragmentation with the one dimensional (1D)-1H-NMR spectra of HPLC-TOFMS-SPE trapped compounds, we elucidated the structures of 36 phenolic conjugates, including the glucuronides of 3’,4’-di, and 3’,4’,5’-trihydroxyphenyl-¿-valerolactone, three urolithin glucuronides, and indole-3-acetic acid glucuronide. We also obtained 26 hours of quantitative excretion profiles for specific valerolactone conjugates. The combination of the HPLC-FTMSn and HPLC-TOFMS-SPE-NMR platforms results in the efficient identification and quantification of low abundant phenolic conjugates down to nanomoles of trapped amounts of metabolite corresponding to micromolar metabolite concentrations in urine
Original languageEnglish
Pages (from-to)7263-7271
JournalAnalytical Chemistry
Volume84
Issue number16
DOIs
Publication statusPublished - 2012

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High pressure liquid chromatography
Metabolites
Nuclear magnetic resonance spectroscopy
Glucuronides
Polyphenols
Bioactivity
Tea

Keywords

  • tandem mass-spectrometry
  • green tea
  • black tea
  • metabolite identification
  • ellagic acid
  • metabolomics
  • polyphenols
  • nmr
  • ingestion
  • phytochemicals

Cite this

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title = "Structural elucidation and quantification of phenolic conjugates present in human urine after tea intake",
abstract = "In dietary polyphenol exposure studies, annotation and identification of urinary metabolites present at low (micromolar) concentrations are major obstacles. In order to determine the biological activity of specific components, it is necessary to have the correct structures and the quantification of the polyphenol-derived conjugates present in the human body. We present a procedure for identification and quantification of metabolites and conjugates excreted in human urine after single bolus intake of black or green tea. A combination of a solid phase extraction (SPE) preparation step and two high pressure liquid chromatography (HPLC)-based analytical platforms was used; namely, accurate mass fragmentation (HPLC-FTMSn) and mass-guided SPE-trapping of selected compounds for nuclear magnetic resonance spectroscopy (NMR) measurements (HPLC-TOFMS-SPE-NMR). HPLC-FTMSn analysis led to the annotation of 138 urinary metabolites, including 48 valerolactone and valeric acid conjugates. By combining the results from MSn fragmentation with the one dimensional (1D)-1H-NMR spectra of HPLC-TOFMS-SPE trapped compounds, we elucidated the structures of 36 phenolic conjugates, including the glucuronides of 3’,4’-di, and 3’,4’,5’-trihydroxyphenyl-¿-valerolactone, three urolithin glucuronides, and indole-3-acetic acid glucuronide. We also obtained 26 hours of quantitative excretion profiles for specific valerolactone conjugates. The combination of the HPLC-FTMSn and HPLC-TOFMS-SPE-NMR platforms results in the efficient identification and quantification of low abundant phenolic conjugates down to nanomoles of trapped amounts of metabolite corresponding to micromolar metabolite concentrations in urine",
keywords = "tandem mass-spectrometry, green tea, black tea, metabolite identification, ellagic acid, metabolomics, polyphenols, nmr, ingestion, phytochemicals",
author = "{van der Hooft}, J.J.J. and {de Vos}, R.C.H. and V. Mihaleva and R.J. Bino and L.O. Ridder and {de Roo}, N. and D.M. Jacobs and {van Duynhoven}, J.P.M. and J.J.M. Vervoort",
year = "2012",
doi = "10.1021/ac3017339",
language = "English",
volume = "84",
pages = "7263--7271",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "16",

}

Structural elucidation and quantification of phenolic conjugates present in human urine after tea intake. / van der Hooft, J.J.J.; de Vos, R.C.H.; Mihaleva, V.; Bino, R.J.; Ridder, L.O.; de Roo, N.; Jacobs, D.M.; van Duynhoven, J.P.M.; Vervoort, J.J.M.

In: Analytical Chemistry, Vol. 84, No. 16, 2012, p. 7263-7271.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Structural elucidation and quantification of phenolic conjugates present in human urine after tea intake

AU - van der Hooft, J.J.J.

AU - de Vos, R.C.H.

AU - Mihaleva, V.

AU - Bino, R.J.

AU - Ridder, L.O.

AU - de Roo, N.

AU - Jacobs, D.M.

AU - van Duynhoven, J.P.M.

AU - Vervoort, J.J.M.

PY - 2012

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N2 - In dietary polyphenol exposure studies, annotation and identification of urinary metabolites present at low (micromolar) concentrations are major obstacles. In order to determine the biological activity of specific components, it is necessary to have the correct structures and the quantification of the polyphenol-derived conjugates present in the human body. We present a procedure for identification and quantification of metabolites and conjugates excreted in human urine after single bolus intake of black or green tea. A combination of a solid phase extraction (SPE) preparation step and two high pressure liquid chromatography (HPLC)-based analytical platforms was used; namely, accurate mass fragmentation (HPLC-FTMSn) and mass-guided SPE-trapping of selected compounds for nuclear magnetic resonance spectroscopy (NMR) measurements (HPLC-TOFMS-SPE-NMR). HPLC-FTMSn analysis led to the annotation of 138 urinary metabolites, including 48 valerolactone and valeric acid conjugates. By combining the results from MSn fragmentation with the one dimensional (1D)-1H-NMR spectra of HPLC-TOFMS-SPE trapped compounds, we elucidated the structures of 36 phenolic conjugates, including the glucuronides of 3’,4’-di, and 3’,4’,5’-trihydroxyphenyl-¿-valerolactone, three urolithin glucuronides, and indole-3-acetic acid glucuronide. We also obtained 26 hours of quantitative excretion profiles for specific valerolactone conjugates. The combination of the HPLC-FTMSn and HPLC-TOFMS-SPE-NMR platforms results in the efficient identification and quantification of low abundant phenolic conjugates down to nanomoles of trapped amounts of metabolite corresponding to micromolar metabolite concentrations in urine

AB - In dietary polyphenol exposure studies, annotation and identification of urinary metabolites present at low (micromolar) concentrations are major obstacles. In order to determine the biological activity of specific components, it is necessary to have the correct structures and the quantification of the polyphenol-derived conjugates present in the human body. We present a procedure for identification and quantification of metabolites and conjugates excreted in human urine after single bolus intake of black or green tea. A combination of a solid phase extraction (SPE) preparation step and two high pressure liquid chromatography (HPLC)-based analytical platforms was used; namely, accurate mass fragmentation (HPLC-FTMSn) and mass-guided SPE-trapping of selected compounds for nuclear magnetic resonance spectroscopy (NMR) measurements (HPLC-TOFMS-SPE-NMR). HPLC-FTMSn analysis led to the annotation of 138 urinary metabolites, including 48 valerolactone and valeric acid conjugates. By combining the results from MSn fragmentation with the one dimensional (1D)-1H-NMR spectra of HPLC-TOFMS-SPE trapped compounds, we elucidated the structures of 36 phenolic conjugates, including the glucuronides of 3’,4’-di, and 3’,4’,5’-trihydroxyphenyl-¿-valerolactone, three urolithin glucuronides, and indole-3-acetic acid glucuronide. We also obtained 26 hours of quantitative excretion profiles for specific valerolactone conjugates. The combination of the HPLC-FTMSn and HPLC-TOFMS-SPE-NMR platforms results in the efficient identification and quantification of low abundant phenolic conjugates down to nanomoles of trapped amounts of metabolite corresponding to micromolar metabolite concentrations in urine

KW - tandem mass-spectrometry

KW - green tea

KW - black tea

KW - metabolite identification

KW - ellagic acid

KW - metabolomics

KW - polyphenols

KW - nmr

KW - ingestion

KW - phytochemicals

U2 - 10.1021/ac3017339

DO - 10.1021/ac3017339

M3 - Article

VL - 84

SP - 7263

EP - 7271

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 16

ER -