Thin-layer chromatography (TLC) is used for the separation and quantification of analytes in the form of a mixture of components in the mobile phase. Nowadays, the HPLC chromatography technique is broadly used for quantification and separation of the components.
TLC as an important and broadly used separation technique as of its complementary features to the HPLC technique. Most TLC applications for the separation apply normal phase methods, while HPLC has reversed-phase methods dominated.
Compared to HPLC, some of the important features of TLC are mentioned here.
• Simple sample preparation
Sample preparation in TLC is simple as compared to HPLC chromatography. Because each sample is separated on a freshly prepared stationary phase TLC required minimum cleaning steps, not including cross-contaminated. Even, solid particles or firmly absorbed impurities in the sample are also not of much concern. This will be a disaster in HPLC separation, decay, leading to column buildup, and ultimately destroying the performance of the column.
• Separate a large number of components
It is perfect to separate the mixture with a large number of components, Apart from this; it is helpful to verify the purity of the components, although hyphenated HPLC can work as several separation techniques.
• The open set-up of the stationary phase and analysis of the whole sample
The mixture is applied in the stationary phase after development, and this is an open system for the separation to the detection of analytes as compare to TLC, HPLC column is a closed system in which a mixture of sample component is brought into the mobile phase and solutes are eluted and detects at different time intervals as per their polarity.
There are times when TLC discloses new and unforeseen information regarding the analytes, while the information in HPLC has been lost by retention at the column, as there is early detection or elution of components and impurities, as well, TLC has less contamination with a stationary phase which is disposable, whereas the column is repeatedly used in HPLC.
• High sample throughput
TLC contains high sample throughput and less time consuming with low cost as well as independent applications and separation, along with many samples. HPLC does not compete with the TLC in the context of the many sample analyses in a given period of time.
• General and specific detection methods
In TLC, Detection of analytes under the short and long-wave UV light even non-chromophores drug can be possible on a developed TLC plate, but traditional and regular HPLCs still use UV detectors; this detector cannot detect all the eluents from the column separation.
• Flexible and versatile dissolving mobile phase and solvent
The selection of the sample solvent is not equally important in TLC to compare to HPLC, as it is removed by evaporation before the development of the plate. In contrast, the selected dissolving solvent in HPLC should be compatible with the mobile phase and column. The same reason is relevant to the mobile phase of TLC, which is fully evaporated before the detection.