title: Solid-phase microextraction to determine micropollutant-macromolecule partition coefficients.
abstract: Aqueous micropollutants such as estradiol can have a large environmental impact-even at low concentrations. Part of understanding this impact involves determining the extent to which the micropollutants interact with macromolecules in water. In environmental samples, relevant macromolecules to which micropollutants bind are referred to as dissolved organic matter, and the most common examples of these in freshwater and coastal seawater are fulvic and humic acids. In living organisms, the most common macromolecules that affect bioavailability of a drug (or toxin) are proteins such as albumin. Using [2, 4, 6, 7 - (3)H]estradiol as an example compound, this protocol uses solid-phase microextraction and scintillation detection as analytical tools to quantify the amount of radiolabeled micropollutant available in solution. The measured free concentration after exposure to various concentrations of macromolecule (dissolved organic matter or protein) or micropollutant is used to determine the partition coefficient in the case of micropollutant-macromolecule interactions. The calibration and preparatory studies take at least 8 d, and the steps to determine the partition coefficient can be completed within 3 d. The protocol could be modified such that nonlabeled compounds are studied; instead of detection of activity by a liquid scintillation counter (LSC), the compounds can be quantified using gas chromatography-mass spectrometry (GC-MS) or liquid chromatography (LC)-MS(/MS).
authors.affiliation: Institute of Biological Chemistry, Biophysics and Bioengineering, Heriot-Watt University, Edinburgh, UK.
authors.forename: Helen L
authors.initials: HL
authors.lastname: Bridle
authors.affiliation: The National Institute of Public Health and the Environment (RIVM), Bilthoven, the Netherlands.
authors.forename: Minne B
authors.initials: MB
authors.lastname: Heringa
authors.affiliation: Department of Membrane Technology, Institute of Functional Interfaces, Karlsruhe Institute of Technology, Karlsruhe, Germany.
authors.forename: Andrea I
authors.initials: AI
authors.lastname: Schäfer
mesh_terms: D000818:Animals; D002417:Cattle; D004958:Estradiol / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry*; D052617:Solid Phase Microextraction / Q000379:methods*; D014867:Water / Q000737:chemistry; D014874:Water Pollutants, Chemical / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D046911:Macromolecular Substances; D014874:Water Pollutants, Chemical; D014867:Water; D004958:Estradiol