Receptor-based high-throughput screening and identification of estrogens in dietary supplements using bioaffinity liquid-chromatography ion mobility mass spectrometry

P. Aqai, N. Gómez Blesa, H. Major, P. Pedotti, L. Varani, V.E.V. Ferrero, W. Haasnoot, M.W.F. Nielen

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

A high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening and identification of recombinant human estrogen receptor a (ERa) ligands in dietary supplements. For screening, a semi-automated mass spectrometric ligand binding assay was developed applying 13C2,15¿N-tamoxifen as non-radioactive label and fast ultra-high-performance–liquid chromatography–electrospray ionisation–triple-quadrupole-MS (UPLC-QqQ-MS), operated in the single reaction monitoring mode, as a readout system. Binding of the label to ERa-coated paramagnetic microbeads was inhibited by competing estrogens in the sample extract yielding decreased levels of the label in UPLC-QqQ-MS. The label showed high ionisation efficiency in positive electrospray ionisation (ESI) mode, so the developed BioMS approach is able to screen for estrogens in dietary supplements despite their poor ionisation efficiency in both positive and negative ESI modes. The assay was performed in a 96-well plate, and all these wells could be measured within 3 h. Estrogens in suspect extracts were identified by full-scan accurate mass and collision-cross section (CCS) values from a UPLC-ion mobility-Q-time-of-flight-MS (UPLC-IM-Q-ToF-MS) equipped with a novel atmospheric pressure ionisation source. Thanks to the novel ion source, this instrument provided picogram sensitivity for estrogens in the negative ion mode and an additional identification point (experimental CCS values) next to retention time, accurate mass and tandem mass spectrometry data. The developed combination of bioaffinity screening with UPLC-QqQ-MS and identification with UPLC-IM-Q-ToF-MS provides an extremely powerful analytical tool for early warning of ERa bioactive compounds in dietary supplements as demonstrated by analysis of selected dietary supplements in which different estrogens were identified.
Original languageEnglish
Pages (from-to)9427-9436
JournalAnalytical and Bioanalytical Chemistry
Volume405
Issue number29
DOIs
Publication statusPublished - 2013

Fingerprint

Dietary supplements
Liquid chromatography
Dietary Supplements
Affinity Chromatography
Liquid Chromatography
Mass spectrometry
Mass Spectrometry
Screening
Estrogens
Throughput
Ions
Labels
Estrogen Receptors
Electrospray ionization
Ion sources
Ionization
Assays
Ligands
Readout systems
Forensic Anthropology

Keywords

  • ms-binding assays
  • multi-residue method
  • anabolic-steroids
  • lc-ms
  • nutritional supplements
  • chemical derivatization
  • native marker
  • contamination
  • transporter
  • validation

Cite this

Aqai, P. ; Gómez Blesa, N. ; Major, H. ; Pedotti, P. ; Varani, L. ; Ferrero, V.E.V. ; Haasnoot, W. ; Nielen, M.W.F. / Receptor-based high-throughput screening and identification of estrogens in dietary supplements using bioaffinity liquid-chromatography ion mobility mass spectrometry. In: Analytical and Bioanalytical Chemistry. 2013 ; Vol. 405, No. 29. pp. 9427-9436.
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abstract = "A high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening and identification of recombinant human estrogen receptor a (ERa) ligands in dietary supplements. For screening, a semi-automated mass spectrometric ligand binding assay was developed applying 13C2,15¿N-tamoxifen as non-radioactive label and fast ultra-high-performance–liquid chromatography–electrospray ionisation–triple-quadrupole-MS (UPLC-QqQ-MS), operated in the single reaction monitoring mode, as a readout system. Binding of the label to ERa-coated paramagnetic microbeads was inhibited by competing estrogens in the sample extract yielding decreased levels of the label in UPLC-QqQ-MS. The label showed high ionisation efficiency in positive electrospray ionisation (ESI) mode, so the developed BioMS approach is able to screen for estrogens in dietary supplements despite their poor ionisation efficiency in both positive and negative ESI modes. The assay was performed in a 96-well plate, and all these wells could be measured within 3 h. Estrogens in suspect extracts were identified by full-scan accurate mass and collision-cross section (CCS) values from a UPLC-ion mobility-Q-time-of-flight-MS (UPLC-IM-Q-ToF-MS) equipped with a novel atmospheric pressure ionisation source. Thanks to the novel ion source, this instrument provided picogram sensitivity for estrogens in the negative ion mode and an additional identification point (experimental CCS values) next to retention time, accurate mass and tandem mass spectrometry data. The developed combination of bioaffinity screening with UPLC-QqQ-MS and identification with UPLC-IM-Q-ToF-MS provides an extremely powerful analytical tool for early warning of ERa bioactive compounds in dietary supplements as demonstrated by analysis of selected dietary supplements in which different estrogens were identified.",
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Receptor-based high-throughput screening and identification of estrogens in dietary supplements using bioaffinity liquid-chromatography ion mobility mass spectrometry. / Aqai, P.; Gómez Blesa, N.; Major, H.; Pedotti, P.; Varani, L.; Ferrero, V.E.V.; Haasnoot, W.; Nielen, M.W.F.

In: Analytical and Bioanalytical Chemistry, Vol. 405, No. 29, 2013, p. 9427-9436.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Receptor-based high-throughput screening and identification of estrogens in dietary supplements using bioaffinity liquid-chromatography ion mobility mass spectrometry

AU - Aqai, P.

AU - Gómez Blesa, N.

AU - Major, H.

AU - Pedotti, P.

AU - Varani, L.

AU - Ferrero, V.E.V.

AU - Haasnoot, W.

AU - Nielen, M.W.F.

PY - 2013

Y1 - 2013

N2 - A high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening and identification of recombinant human estrogen receptor a (ERa) ligands in dietary supplements. For screening, a semi-automated mass spectrometric ligand binding assay was developed applying 13C2,15¿N-tamoxifen as non-radioactive label and fast ultra-high-performance–liquid chromatography–electrospray ionisation–triple-quadrupole-MS (UPLC-QqQ-MS), operated in the single reaction monitoring mode, as a readout system. Binding of the label to ERa-coated paramagnetic microbeads was inhibited by competing estrogens in the sample extract yielding decreased levels of the label in UPLC-QqQ-MS. The label showed high ionisation efficiency in positive electrospray ionisation (ESI) mode, so the developed BioMS approach is able to screen for estrogens in dietary supplements despite their poor ionisation efficiency in both positive and negative ESI modes. The assay was performed in a 96-well plate, and all these wells could be measured within 3 h. Estrogens in suspect extracts were identified by full-scan accurate mass and collision-cross section (CCS) values from a UPLC-ion mobility-Q-time-of-flight-MS (UPLC-IM-Q-ToF-MS) equipped with a novel atmospheric pressure ionisation source. Thanks to the novel ion source, this instrument provided picogram sensitivity for estrogens in the negative ion mode and an additional identification point (experimental CCS values) next to retention time, accurate mass and tandem mass spectrometry data. The developed combination of bioaffinity screening with UPLC-QqQ-MS and identification with UPLC-IM-Q-ToF-MS provides an extremely powerful analytical tool for early warning of ERa bioactive compounds in dietary supplements as demonstrated by analysis of selected dietary supplements in which different estrogens were identified.

AB - A high-throughput bioaffinity liquid chromatography-mass spectrometry (BioMS) approach was developed and applied for the screening and identification of recombinant human estrogen receptor a (ERa) ligands in dietary supplements. For screening, a semi-automated mass spectrometric ligand binding assay was developed applying 13C2,15¿N-tamoxifen as non-radioactive label and fast ultra-high-performance–liquid chromatography–electrospray ionisation–triple-quadrupole-MS (UPLC-QqQ-MS), operated in the single reaction monitoring mode, as a readout system. Binding of the label to ERa-coated paramagnetic microbeads was inhibited by competing estrogens in the sample extract yielding decreased levels of the label in UPLC-QqQ-MS. The label showed high ionisation efficiency in positive electrospray ionisation (ESI) mode, so the developed BioMS approach is able to screen for estrogens in dietary supplements despite their poor ionisation efficiency in both positive and negative ESI modes. The assay was performed in a 96-well plate, and all these wells could be measured within 3 h. Estrogens in suspect extracts were identified by full-scan accurate mass and collision-cross section (CCS) values from a UPLC-ion mobility-Q-time-of-flight-MS (UPLC-IM-Q-ToF-MS) equipped with a novel atmospheric pressure ionisation source. Thanks to the novel ion source, this instrument provided picogram sensitivity for estrogens in the negative ion mode and an additional identification point (experimental CCS values) next to retention time, accurate mass and tandem mass spectrometry data. The developed combination of bioaffinity screening with UPLC-QqQ-MS and identification with UPLC-IM-Q-ToF-MS provides an extremely powerful analytical tool for early warning of ERa bioactive compounds in dietary supplements as demonstrated by analysis of selected dietary supplements in which different estrogens were identified.

KW - ms-binding assays

KW - multi-residue method

KW - anabolic-steroids

KW - lc-ms

KW - nutritional supplements

KW - chemical derivatization

KW - native marker

KW - contamination

KW - transporter

KW - validation

U2 - 10.1007/s00216-013-7384-1

DO - 10.1007/s00216-013-7384-1

M3 - Article

VL - 405

SP - 9427

EP - 9436

JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

SN - 1618-2642

IS - 29

ER -