Triple Bioaffinity Mass Spectrometry Concept for Thyroid Transporter Ligands

P. Aqai, C. Fryganas, M. Mizuguchi, W. Haasnoot, M.W.F. Nielen

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)

Abstract

For the analysis of thyroid transporter ligands, a triple bioaffinity mass spectrometry (BioMS) concept was developed, with the aim at three different analytical objectives: rapid screening of any ligand, confirmation of known ligands in accordance with legislative requirements, and identification of emerging yet unknown ligands. These three purposes share the same biorecognition element, recombinant thyroid transport protein transthyretin (rTTR), and dedicated modes of liquid chromatography-mass spectrometry (LC-MS). For screening, a rapid and radiolabel-free competitive inhibition MS binding assay was developed with fast ultrahigh performance-liquid chromatography-electrospray ionization-triple-quadrupole-MS (UPLC-QqQ-MS) as the readout system. It uses the nonradioactive stable isotopic thyroid hormone 13C6-l-thyroxine as the label of which the binding to rTTR is inhibited by any ligand such as thyroid drugs and thyroid endocrine disrupting chemicals (EDCs). To this end, rTTR is either used in solution or immobilized on paramagnetic microbeads. The concentration-dependent inhibition of the label by the natural thyroid hormone l-thyroxine (T4), as a model analyte, is demonstrated in water at part-per-trillion and in urine at part-per-billion level. For confirmation of identity of known ligands, rTTR was used for bioaffinity purification for confirmation of naturally present free T4 in urine. As a demonstrator for identification of unknown ligands, the same rTTR was used again but in combination with nano-UPLC-quadrupole time-of-flight-MS (nLC-Q-TOF-MS) and urine samples spiked with the model “unknown” EDCs triclosan and tetrabromobisphenol-A. This study highlights the potential of BioMS using one affinity system, both for rapid screening and for confirmation and identification of known and unknown emerging thyroid EDCs.
Original languageEnglish
Pages (from-to)6488-6493
Number of pages6
JournalAnalytical Chemistry
Volume84
DOIs
Publication statusPublished - 2012

Fingerprint

Mass spectrometry
Ligands
Endocrine Disruptors
Screening
Liquid chromatography
Thyroxine
Thyroid Hormones
Labels
Triclosan
Readout systems
Electrospray ionization
Prealbumin
Purification
Assays
Carrier Proteins
Water
Pharmaceutical Preparations

Keywords

  • brominated flame retardants
  • ms-binding assays
  • human transthyretin
  • in-vitro
  • tetrabromobisphenol-a
  • disrupting chemicals
  • surface-water
  • pharmaceuticals
  • identification
  • metabolites

Cite this

Aqai, P. ; Fryganas, C. ; Mizuguchi, M. ; Haasnoot, W. ; Nielen, M.W.F. / Triple Bioaffinity Mass Spectrometry Concept for Thyroid Transporter Ligands. In: Analytical Chemistry. 2012 ; Vol. 84. pp. 6488-6493.
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Triple Bioaffinity Mass Spectrometry Concept for Thyroid Transporter Ligands. / Aqai, P.; Fryganas, C.; Mizuguchi, M.; Haasnoot, W.; Nielen, M.W.F.

In: Analytical Chemistry, Vol. 84, 2012, p. 6488-6493.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Triple Bioaffinity Mass Spectrometry Concept for Thyroid Transporter Ligands

AU - Aqai, P.

AU - Fryganas, C.

AU - Mizuguchi, M.

AU - Haasnoot, W.

AU - Nielen, M.W.F.

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AB - For the analysis of thyroid transporter ligands, a triple bioaffinity mass spectrometry (BioMS) concept was developed, with the aim at three different analytical objectives: rapid screening of any ligand, confirmation of known ligands in accordance with legislative requirements, and identification of emerging yet unknown ligands. These three purposes share the same biorecognition element, recombinant thyroid transport protein transthyretin (rTTR), and dedicated modes of liquid chromatography-mass spectrometry (LC-MS). For screening, a rapid and radiolabel-free competitive inhibition MS binding assay was developed with fast ultrahigh performance-liquid chromatography-electrospray ionization-triple-quadrupole-MS (UPLC-QqQ-MS) as the readout system. It uses the nonradioactive stable isotopic thyroid hormone 13C6-l-thyroxine as the label of which the binding to rTTR is inhibited by any ligand such as thyroid drugs and thyroid endocrine disrupting chemicals (EDCs). To this end, rTTR is either used in solution or immobilized on paramagnetic microbeads. The concentration-dependent inhibition of the label by the natural thyroid hormone l-thyroxine (T4), as a model analyte, is demonstrated in water at part-per-trillion and in urine at part-per-billion level. For confirmation of identity of known ligands, rTTR was used for bioaffinity purification for confirmation of naturally present free T4 in urine. As a demonstrator for identification of unknown ligands, the same rTTR was used again but in combination with nano-UPLC-quadrupole time-of-flight-MS (nLC-Q-TOF-MS) and urine samples spiked with the model “unknown” EDCs triclosan and tetrabromobisphenol-A. This study highlights the potential of BioMS using one affinity system, both for rapid screening and for confirmation and identification of known and unknown emerging thyroid EDCs.

KW - brominated flame retardants

KW - ms-binding assays

KW - human transthyretin

KW - in-vitro

KW - tetrabromobisphenol-a

KW - disrupting chemicals

KW - surface-water

KW - pharmaceuticals

KW - identification

KW - metabolites

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DO - 10.1021/ac300543u

M3 - Article

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SP - 6488

EP - 6493

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

ER -