**7.2 High performance thin layer chromatography (HPTLC)**

 HPTLC is a more powerful separation tool for quantitative analysis and it uses the technique in a more optimized way. High performance thin layer chromatography (HPTLC) is based on the principle of planar chromatography where separation of sample components is achieved on high performance layers with detection and data evaluation. These high performance layers on TLC plates are precoated with an adsorbent of 6 micron particle size and a 160 microns layer thickness. The lesser the thickness of layer and particle size results in increased plate efficiency as well as nature of separation. HPTLC has an ability to show its performance on graphical representation in the form of chromatogram. Separation can be easily visualized by pictorial representation, which is possible only in case of HPTLC. The procedure used is as follows: silica gel 60 F254 precoated plates (20 × 10 cm) are used with any developed solvent system. Different extracts are to be spotted on precoated HPTLC plates. Spots of different concentration (l μL) was applied on HPTLC plates to study the exact separation of spots. Saturation time will be 20 min and room temperature 25°C ± 2°C. TLC plates were developed up to 8 cm. After air drying, a plate was heated at 110°C for 2–3 min. In TLC fingerprinting analysis, the information can be stored and recorded using specific highly sophisticated instruments like high performance TLC scanner. It gives information about the chromatogram, retardation factor (Rf) values, the color of the separated bands, their absorption spectra, and λ max. After derivatization and using different visualization reagents, snaps of TLC plates can be obtained and saved for further process. Thus, this represents TLC fingerprint profile of the provided sample. The information so generated has


#### **Table 1.**

*TLC mobile phase for important classes of phytoconstituents [15].* 

a potential application in the identification of an authentic drug when compared with bioactive marker and it helps in maintaining the quality and consistency of the isolates or herbal drugs in natural drug discovery [18].
