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Updated: Dec 21, 2025

Radiosynthesis, Quality Control, and Small Animal Positron Emission Tomography Imaging of 68Ga-Labelled Nano Molecules

Radiosynthesis, Quality Control, and Small Animal Positron Emission Tomography Imaging of ⁶⁸Ga-Labelled Nano Molecules

Published on: October 4, 2024

New sensitive method for HEPES quantification in ⁶⁸Ga-radiopharmaceuticals.

I F Antunes¹, G M Franssen², R Zijlma³

¹Department of Nuclear Medicine and Molecular Imaging, University of Groningen, University Medical Center Groningen, Hanzeplein 1, 9713GZ, Groningen, The Netherlands. i.farinha.antunes@umcg.nl.

EJNMMI Radiopharmacy and Chemistry

|May 16, 2020

Summary

A new HPLC method accurately quantifies 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) in ⁶⁸Ga-radiopharmaceuticals, unlike less sensitive TLC methods. This ensures patient safety by preventing excessive HEPES doses.

Keywords:

68Ga-radiopharmaceuticals HEPES HPLC and quality control

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Area of Science:

Nuclear Medicine
Radiopharmaceutical Chemistry
Analytical Chemistry

Background:

GMP-certified ⁶⁸Ga generators have increased ⁶⁸Ga-radiopharmaceutical use.
2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) buffer improves radiosynthesis but is an impurity.
European Pharmacopoeia (Ph Eur) limits HEPES to 200 μg/V_Injected for radiopharmaceuticals like edotreotide.

Purpose of the Study:

To evaluate the sensitivity of the Ph Eur thin-layer chromatography (TLC) method for HEPES quantification.
To develop and validate a new high-performance liquid chromatography (HPLC) method for HEPES quantification in ⁶⁸Ga-radiopharmaceuticals.
To compare the new HPLC method with the Ph Eur TLC method and assess HEPES levels in various ⁶⁸Ga-radiopharmaceuticals from different institutes.

Main Methods:

Tested the Ph Eur TLC method for HEPES detection in ⁶⁸Ga-radiopharmaceuticals.
Developed and validated a modified TLC method and a new HPLC method for HEPES quantification.
Analyzed ⁶⁸Ga-radiopharmaceutical samples from multiple institutions using the developed methods.

Main Results:

The Ph Eur TLC method failed to detect HEPES within 4 minutes of iodine incubation.
The modified TLC method required 2 hours for visible spots, only at concentrations >1 mg/mL.
The HPLC method achieved a limit of quantification (LOQ) of 3 μg/mL and a limit of detection (LOD) of 1 μg/mL, detecting excessive HEPES in [⁶⁸Ga]Ga-NODAGA-Exendin samples.

Conclusions:

The Ph Eur and modified TLC methods are insufficiently sensitive for quality control (QC) of HEPES in ⁶⁸Ga-radiopharmaceuticals.
The new HPLC method is sensitive, quantitative, reproducible, and suitable for QC release.
This HPLC method identified ⁶⁸Ga-radiopharmaceutical batches exceeding the pharmacopoeial HEPES limit, enabling dose adjustment for patient safety.