posted on 2022-10-19, 20:03authored byTakuya Kubo, Shuhei Yamamoto, Yoshiyuki Watabe, Tetsuya Tanigawa, Daisuke Nakajima, Koji Otsuka
To date, an identification protocol for endocrine disruptors
that
bind to the thyroid hormone receptor (TR) has not been established.
A method for screening and identifying TR-binding substances is highly
required due to the existence of unknown TR-binding substances from
the environment. Here, we conceived a chromatographic method using
a molecularly imprinted polymer (MIP) to create a novel screening
protocol for the endocrine disruptors. A receptor-imitating MIP was
prepared using N-acetylthyroxine (AcetylT4) and 4-vinylpyridine
as a pseudo-template and a functional monomer, respectively, based
on the existing molecular recognition mechanism of the TR. The receptor-imitating
MIP provided molecular recognition ability for all the TR-binding
substances that were employed in this study. The prepared MIPs were
packed into a high-performance liquid chromatography column for the
simultaneous analysis of TR-binding and non-binding substances. The
former was strongly retained, while the latter was not. The presence
or absence of TR-binding/non-binding activity resulted in successful
dichotomous separation. Additionally, the surface imprinting technique
was applied to improve the separation performance of the MIP packing
material. MIP-coated uniformly sized silica-based particles of 5 μm
were successfully prepared, and the MIP-coated silica column enabled
more efficient dichotomous separation of TR-binding and non-binding
substances.