- HPLC and HPTLC are two types of automated separation techniques with similar principles of separation. Furthermore, HPLC consists of a pump-driven flow system through the stationary phase filled in a column while HPTLC is a type of planar chromatography in which the solvent moves through a stationary phase fixed on a plate. Moreover, HPLC consists of a closed system, while HPTLC is an open system.
- Both HPLC and HPTLC are the separation techniques of chromatography operating on the same principles of separation.
- HPLC is known as high-pressure liquid chromatography and is also known as High-performance liquid chromatography, it is an analytical technique, that is used to separate, determine, and identify each component in a complex mixture.
- It depends on a pressurized mobile phase provided by the pumps in which the sample mixture is included and passes through the HPLC column. Every analyte in the complex mixture interacts with the absorbing material in a different way, which has dissimilar flow rates for the different analytes and leads to the separation of the analyte or components.
- High-performance thin-layer chromatography is the advanced form of TLC and contains the use of chromatographic layers of cutting-edge separation efficiency for all stages of the process and the use of state-of-the-art instrumentation, reproducible chromatogram, accurate sample application, and software-based evaluation.
- HPTLC is a complete concept in which a broad, standardized method depends on scientific facts, and the quantitative and qualitative analysis uses validated methods.
- The principle of HPTLC has a similar approach and employs the same physical principles of TLC (adsorption chromatography) i.e. the principle of separation is adsorption. The mobile phase solvent flows through because of capillary action. The components move according to their affinities towards the adsorbent.
- HPTLC (high-performance thin-layer chromatography) is a sophisticated form of TLC, which provides superior separation efficiency. The HPTLC concept includes validated methods for qualitative and quantitative analysis and fulfills all quality requirements for use in fully regulated environments.
- The CAMAG TLC Scanner 4 (densitometer) is the most advanced workstation for densitometric evaluation of TLC/HPTLC chromatograms and other planar objects. The chromatogram is evaluated under white or UV light.
- HPLC is a highly improved form of column chromatography. A pump forces a solvent through a column under high pressures of up to 400 atmospheres. The column packing material or adsorbent or stationary phase is typically a granular material made of solid particles such as silica or polymers.
- HPLC is used to split a mixture of compounds in the fields of analytical chemistry, biochemistry, Pharmaceutical, and industrial. The main purposes of using HPLC are for identifying, quantifying, and purifying the individual components of the mixture.
- Water purification.
- Detection of impurities in pharmaceutical industries.
- Pre-concentration of trace components.
- Ligand-exchange chromatography.
Parameters | HPLC | HPTLC |
Type | Reverse Phase Chromatography | Straight Phase Chromatography |
Stationary phase | Liquid | Solid |
Conditioning phase | None | Gas |
Separation by | Partition | Adsorption |
Results | By machine | By machine + eyes |
Analysis | On ‐ line | Offline |
Resolution | Very high | Moderate to high |
Chromatography System | Closed | Open |
Separating medium | Tubular column | Planar layer (plate) |
Strongly Retarded Fractions Seen As | Broad peaks | Sharp Peaks |
Analysis in parallel | No. Only 1 at a time | Yes. Upto 100 samples. |
High temp. / pressure | High pressure | None |
Time per sample | 2‐ 60 min | 1-30 min |
Data obtained from chromatography | Limited to very high | High to very high |
Post chromatography derivatization | Limited possibilities. Cumbersome. | Simple. Possible for every sample. Gives additional information |
Fraction collection / micro preparative chromatography | Requires prep. scale chromatography & fraction collector | Simple. No special requirements |
Sensitivity | High to ultra | high Moderate to ultrahigh |
Fluorescence data | Possible, optional | Possible, built-in |
Detectors | UV, Fluor, Electrochem Light scatter, MS | UV ‐ Vis, bioluminescence, MS |
Chromatogram image documentation | No | Yes. At 254 & 366 nm & visible |
Sample clean-up | Through Column reusable | Not so imp. Layer disposable |
Chromatographic fingerprint | Yes, but limited | Yes. Comprehensive |
Cost per analysis | Very high | Low |
Eqpt. maintenance | Very high | Low |
Analyst’s skills required | High to very high | Low (TLC) to high (HPTLC) |
