The HPTLC is an effective tool to verify the fingerprint of identity of plant materials against a reference plate. We are able to confirm the profile of a plant thanks to this tool, and to ensure the right material is being used.
With an HPTLC, individual components of a mixture are separated on a thin glass-coated plate, which is then placed in a developing chamber. The TLC plate is placed under an ultraviolet lamp, and bands of the different components are visualized. Unlike a standard TLC, with an HPTLC many tasks are automated via robotics, eliminating uncertainty from samples being applied to plates by hand.
We use the HPTLC to detect contamination by mycotoxins, a dangerous class of toxins that can develop on plant matter in humid conditions.
You can see an HPTLC plate below.
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Thin-Layer Chromatography has been a well-known method for analysis of botanicals and other complex samples since the end of the 1930s. Today’s modern thin-layer chromatography combines the advantage of analytical robustness and high sample throughput with the possibility to use all kind of specific detection methods, e.g. classical UV/Vis/Fluorescence detection, mass spectrometry or effect-directed analysis. This method increases the amount of information for a fast and efficient screening for new compounds and the identification of raw materials especially for samples with a high matrix load such as herbal drugs, cosmetic and food samples.
HPTLC, the most advanced form of Thin-Layer Chromatography, is a powerful yet simple and cost effective tool for testing identity, purity, and strength (content) of botanicals as well as excluding adulteration during quality control. With the publication of general chapters by the United States Pharmacopoeia (USP <203>) and European Pharmacopoeia (Ph.Eur. 2.8.25) HPTLC has officially come into existence as a highly standardized and therefore reproducible analytical technique. The use of high performance plates, suitable instrument and software, a standardized methodology, and validated methods ensures reliable results that are fully compliant with current Good Manufacturing Practice (cGMP). HPTLC fingerprints allow convenient visual comparison of multiple samples even if those originate from different plates (and different laboratories worldwide). Reference images (HPTLC fingerprints of botanical reference materials or other references) can be used to qualify data and pass/fail samples based on similarity or difference.
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High-Performance Thin-Layer Chromatography (HPTLC) is an analytical technique used for the separation, identification, and quantification of components in a mixture. It is a powerful and versatile chromatographic method that shares similarities with traditional thin-layer chromatography (TLC) but offers improved performance in terms of speed, sensitivity, and resolution. Here are some key aspects of HPTLC as an analytical tool:
- Principle:
- HPTLC operates on the same principles as TLC, involving the separation of components based on their affinity for a stationary phase and a mobile phase.
- The stationary phase is typically a thin layer of adsorbent material (e.g., silica gel or aluminum oxide) coated on a glass or aluminum plate.
- The mobile phase, a solvent or a mixture of solvents, moves up the plate through capillary action, carrying the sample with it.
- Advantages of HPTLC:
- High Resolution: HPTLC provides better resolution and separation of components compared to traditional TLC.
- Speed: HPTLC allows for faster separations, making it a time-efficient technique.
- Sensitivity: It offers improved sensitivity, enabling the detection of trace amounts of analytes.
- Quantification: HPTLC can be used for quantitative analysis by densitometry, where the separated spots are quantified based on their intensity.
- Instrumentation:
- HPTLC systems include a development chamber, where the chromatographic separation takes place.
- Automated sample application and development systems are available for increased precision and reproducibility.
- Densitometers are used for the quantitative analysis of separated components.
- Applications:
- HPTLC finds applications in various industries, including pharmaceuticals, food and beverages, environmental analysis, forensic science, and more.
- It is particularly useful for analyzing complex mixtures and identifying impurities in pharmaceuticals.
- Quantitative Analysis:
- HPTLC can be used for quantitative analysis by measuring the intensity of spots corresponding to the separated components.
- Calibration curves are often constructed using standard solutions to relate spot intensity to concentration.
- Sample Preparation:
- Proper sample preparation is crucial for successful HPTLC analysis.
- Samples are often applied as small spots or bands on the TLC plate.
- Detection Methods:
- Visualization of separated components is typically done using various detection methods, including UV absorption, fluorescence, and derivatization reactions.
- Densitometry is often employed for quantitative analysis.
- Method Development:
- Optimizing the chromatographic conditions, such as the choice of stationary phase, mobile phase composition, and development chamber conditions, is essential for achieving the desired separation.
In summary, HPTLC is a valuable analytical tool that combines the advantages of thin-layer chromatography with improved performance characteristics. Its high resolution, speed, and sensitivity make it suitable for a wide range of applications in analytical chemistry.
