What is the Difference Between TLC and HPTLC?
🆚 Go to Comparative Table 🆚The main difference between Thin-Layer Chromatography (TLC) and High-Performance Thin-Layer Chromatography (HPTLC) lies in the characteristics of the separation plate and the technology used. Here are the key differences between TLC and HPTLC:
- Particle Size: TLC plates have larger absorbent particles, while HPTLC plates have very minute absorbent particles. The mean particle size for TLC is 10-12 µm, while for HPTLC, it is 5-6 µm.
- Separation Efficiency: HPTLC is an advanced technique compared to TLC, providing superior separation efficiency. HPTLC plates have a higher packing density and a smoother surface, which reduces sample diffusion and results in compact bands or spots.
- Detection Sensitivity: HPTLC has a thinner layer and smaller particle size, significantly increasing detection sensitivity.
- Plate Material: TLC plates can be made of paper, plastic, or glass, but HPTLC plates are based on optimized silica gel 60.
- Sample Volume: The sample volume for HPTLC is typically 0.1-0.5 µl, while for TLC, it is 1-5 µl.
- Detection Limits: HPTLC has lower detection limits for both absorption and fluorescence, making it more sensitive than TLC.
- Automation: Most steps of the HPTLC technique can be automated, while TLC is a more manual process.
- Analysis Time: HPTLC provides faster results compared to TLC.
In summary, HPTLC is an advanced and more efficient version of TLC, offering improved separation efficiency, detection sensitivity, and automation capabilities.
Comparative Table: TLC vs HPTLC
Here is a table comparing the differences between Thin Layer Chromatography (TLC) and High-Performance Thin Layer Chromatography (HPTLC):
Feature | TLC | HPTLC |
---|---|---|
Layer thickness | 250 µm | 200 µm (100 µm) |
Plate height | 30 µm | 12 µm |
Typical migration distance | 10 - 15 cm | 3 - 6 cm |
Typical separation time | 20 - 200 min | 3 - 20 min |
Mean particle size | 10 - 12 µm | 5 - 6 µm |
Particle size distribution | 5 - 20 µm | 4 - 8 µm |
Number of samples per plate | < 10 | < 36 (72) |
Sample volume | 1 - 5 µl | 0.1 - 0.5 µl |
Detection limits: absorption | 1 - 5 ng | 100 - 500 pg |
Detection limits: fluorescence | 50 - 100 pg | 5 - 10 pg |
TLC is a time-consuming technique, while HPTLC provides faster results and higher sensitivity. HPTLC plates are made of optimized silica gel with a smaller particle size, resulting in a smoother surface, reduced sample diffusion, and more compact bands or spots. The smaller particle size and thinner layer in HPTLC also increase detection sensitivity.
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