Effects of Cations on HPTS Fluorescence and Quantification of Free Gadolinium Ions in Solution; Assessment of Intracellular Release of Gd<sup>3+</sup> from Gd-Based MRI Contrast Agents

8-Hydroxypyrene-1,3,6-trisulfonate (HPTS) is a small, hydrophilic fluorescent molecule. Since the pKa of the hydroxyl group is close to neutrality and quickly responds to pH changes, it is widely used as a pH-reporter in cell biology for measurements of intracellular pH. HPTS fluorescence (both exci...

Full description

Bibliographic Details
Main Authors: Angelo Scarciglia, Enza Di Gregorio, Silvio Aime, Giuseppe Ferrauto
Format: Article
Language:English
Published: MDPI AG 2022-04-01
Series:Molecules
Subjects:
Online Access:https://www.mdpi.com/1420-3049/27/8/2490
Description
Summary:8-Hydroxypyrene-1,3,6-trisulfonate (HPTS) is a small, hydrophilic fluorescent molecule. Since the pKa of the hydroxyl group is close to neutrality and quickly responds to pH changes, it is widely used as a pH-reporter in cell biology for measurements of intracellular pH. HPTS fluorescence (both excitation and emission spectra) at variable pH was measured in pure water in the presence of NaCl solution or in the presence of different buffers (PBS or hepes in the presence or not of NaCl) and in a solution containing BSA. pKa values have been obtained from the sigmoidal curves. Herein, we investigated the effect of mono-, di-, and trivalent cations (Na<sup>+</sup>, Ca<sup>2+</sup>, La<sup>3+</sup>, Gd<sup>3+</sup>) on fluorescence changes and proposed its use for the quantification of trivalent cations (e.g., gadolinium ions) present in solution as acqua-ions. Starting from the linear regression, the LoD value of 6.32 µM for the Gd<sup>3+</sup> detection was calculated. The effects on the emission were also analyzed in the presence of a combination of Gd<sup>3+</sup> at two different concentrations and the previously indicated mono and di-valent ions. The study demonstrated the feasibility of a qualitative method to investigate the intracellular Gd<sup>3+</sup> release upon the administration of Gd-based contrast agents in murine macrophages.
ISSN:1420-3049