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Thin-Layer Chromatography (TLC): Overview

Thin-Layer Chromatography (TLC): Overview

Thin-layer chromatography (TLC) is a chromatography technique that separates compounds based on their polarity. TLC typically uses polar silica gel, a form of silicon dioxide, as the stationary phase. The silica gel contains hydroxyl (OH) groups on its surface, which form hydrogen bonds with polar compounds, influencing their adhesion to the stationary phase.
To begin the analysis, a mixture of compounds is spotted on the starting line on the TLC plate using a thin capillary. The bottom of the...

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Related Experiment Video

Updated: Dec 31, 2025

Optimization of Radiochemical Reactions using Droplet Arrays

Optimization of Radiochemical Reactions using Droplet Arrays

Published on: February 12, 2021

High-throughput radio-TLC analysis.

Jia Wang¹, Alejandra Rios², Ksenia Lisova²

¹Crump Institute for Molecular Imaging, University of California Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, University of California Los Angeles, Los Angeles, CA 90095, USA.

Nuclear Medicine and Biology

|January 1, 2020

Summary

High-throughput radio-thin layer chromatography (radio-TLC) using Cerenkov luminescence imaging (CLI) significantly reduces analysis time for radiopharmaceuticals. This method enables rapid purity assessment and reaction optimization, improving efficiency in radiosynthesis.

Keywords:

High-throughput analysis Quality control testing Radiochemical purity Radiopharmaceutical analysis Radiosynthesis optimization Thin-layer chromatography

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Last Updated: Dec 31, 2025

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Published on: February 12, 2021

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Area of Science:

Radiochemistry
Analytical Chemistry
Nuclear Medicine

Background:

Radio-thin layer chromatography (radio-TLC) is a standard technique for assessing radiopharmaceutical purity and optimizing radiosynthesis.
Conventional radio-TLC is time-consuming, especially for analyzing numerous samples during reaction optimization.
Cerenkov luminescence imaging (CLI) has shown potential for reading radio-TLC plates.

Purpose of the Study:

To develop a high-throughput method for radio-TLC analysis using Cerenkov luminescence imaging (CLI).
To demonstrate the application of this method for analyzing multiple radiopharmaceutical samples simultaneously.
To significantly reduce the overall analysis time compared to conventional radio-TLC methods.

Main Methods:

Parallel development of multiple radioactive samples on a single TLC plate.
Simultaneous readout of separated samples using Cerenkov luminescence imaging (CLI).
Image processing using custom MATLAB software to quantify radioactive regions of interest (ROIs).

Main Results:

Reduced total analysis time for eight samples from 48 minutes (conventional) to 7.5 minutes (CLI-based).
Enhanced separation resolution with CLI allowed detection of a low-abundance side product in [18F]FET samples, missed by conventional radio-TLC scanners.
Achieved high labeling efficiency (99%) for [177Lu]Lu-PSMA-617 in 10 minutes, compared to the typical 30 minutes.

Conclusions:

Cerenkov luminescence imaging combined with parallel TLC development offers a practical and rapid approach for high-throughput radio-TLC analysis.
This method significantly accelerates radiopharmaceutical purity assessment and reaction optimization processes.
The enhanced resolution and speed of CLI-based radio-TLC improve analytical capabilities in radiochemistry.