What is HPLC PDF?
High performance liquid chromatography (HPLC) is an important qualitative and quantitative technique, generally used for the estimation of pharmaceutical and biological samples. It is the most versatile, safest, dependable and fastest chromatographic technique for the quality control of drug components.
What is HPTLC principle?
High-Performance Thin-Layer Chromatography (HPTLC) Principle
The HPTLC works on the same principles as TLC such as the principle of separation is adsorption. The mobile phase or solvent flows through the capillary action. The analytes move according to their affinities towards the stationary phase (adsorbent).
What are the steps involved in HPTLC?
The HPTLC suite is composed of a number of components.
- Sample Applicator – Application of sample to stationary phase i.e. TLC/HPTLC plate.
- Development Chamber – Separation of components.
- Derivatizer set and Plate heater – Derivatization: Chemical spraying of plate to detect compounds not visible under white/UV light.
Which instrument is used for HPTLC?
The compounds in the sample will then separate based on their polarities. The difference lies in how the experiment is performed, in that HPTLC uses a computer system connected to an auto-sampler and a scanner (and occasionally an Automatic Developing Chamber and a Chromatogram Immersion Device).
What are the 5 parts of HPLC and their function?
Main High-Performance Liquid Chromatography (HPLC) Components. The HPLC system mainly consists of an infusion pump, a sampler, a chromatographic column, a detector, and a data recording and processing device. Among them, the infusion pump, the chromatographic column, and the detector are key components.
What are the types of HPLC?
HPLC analyses fall into four different categories: (1) reverse phase chromatography where the stationary phase is hydrophobic (C18-modified silica), (2) normal phase chromatography where the stationary phase is hydrophilic (silica), (3) hydrophilic interaction chromatography, a hybrid of techniques 1 and 2, and (4) ion …
Why is HPLC used?
The purpose high performance liquid chromatography (HPLC) analysis of any drugs is to confirm the identity of a drug and provide quantitative results and also to monitor the progress of the therapy of a disease.
Who invented HPTLC?
For the purpose of analyzing ezetimibe in combination with simvastatin, three HPTLC methods have been developed. The first method was developed by Shivshankar et al. [33] and yielded higher Rf values for either ezetimibe or simvastatin. In the method developed by Dhaneshwar et al.
What does Rf value mean?
What is RF Value? The Rf (retardation factor) value is the ratio of the solute’s distance travelled to the solvent’s distance travelled. The word comes from chromatography when it was discovered that a given component will always travel the same distance in a given solvent under the same conditions.
What is the difference between HPLC and HPTLC?
Definition. HPLC refers to a form of liquid chromatography to separate compounds dissolved in a solution, while HPTLC refers to a most advanced form of planar chromatography. Thus, this is the main difference between HPLC and HPTLC.
What is application of HPTLC?
HPTLC is used for purity control of chemicals, pesticides, steroids, and water analysis. [50] HPTLC is also widely used for analysis of vitamins, water-soluble food dyes, pesticides in fruits, vegetables, and other food stuffs.
Is HPTLC quantitative or qualitative?
officinalis extract composition from different parts of the plant. The results found HPTLC to be effective for qualitative analysis, however, HPLC was found to be more accurate for quantitative analysis.
Which column is used in HPLC?
The reversed-phase HPLC column is the most versatile and commonly used column type and can be used for a wide range of different types of analytes.
Which detector is used in HPLC?
UV detector is a very commonly used detector for HPLC analysis. During the analysis, sample goes through a clear color-less glass cell, called flow cell.
Where is HPLC used?
HPLC is the form of liquid chromatography that is generally used in the pharmaceutical industry, as it can provide the precise results that are required. The results can be used to analyse finished drug products and their ingredients quantitatively and qualitatively during the manufacturing process.
Why C18 column is used in HPLC?
Because of the extra carbons, C18 has a larger surface area that the mobile phase has to travel across. This offers more interaction time between the bonded phase and the elutes. Thus the sample elutes more slowly and has more separation.
Why HPLC is called High pressure?
HPLC stands for High Performance Liquid Chromatography.
Its earlier name was High Pressure Liquid Chromatography because it involved use of liquid mobile phase requiring higher pressures than gases used in Gas Chromatography.
Why is Hptlc used?
Why is HPTLC high performance?
HPTLC has been reported to offer several advantages over the more widely used high-performance liquid chromatography (HPLC) technique because of its high throughput (multiple samples can be analyzed simultaneously), economy in use of mobile phase, and cost effectiveness (Mishra et al., 2017a, b).
What is Rf value formula?
The following expression can be used to calculate Rf values: Rf = Distance travelled by the substance from reference line (cm)/Distance travelled by the solvent front from reference line (cm)
What is Rf unit?
Radio frequency is measured in units called hertz (Hz), which represent the number of cycles per second when a radio wave is transmitted. One hertz equals one cycle per second; radio waves range from thousands (kilohertz) to millions (megahertz) to billions (gigahertz) of cycles per second.
How is HPTLC better than TLC?
HPTLC plates are based on optimized silica gel 60 with a significantly smaller particle size than used for classical TLC. This allows a higher packing density and a smoother surface. Hence, sample diffusion is reduced, resulting in compact bands or spots.
Which is better HPTLC or HPLC?
Chromatography techniques are commonly used to produce a chemical fingerprint of a plant. HPTLC is effective tool in qualitative analysis, while HPLC is more accurate. HPTLC combined with HPLC allows rapid analysis while maintaining accuracy.
When was HPTLC invented?
In the mid-1960s, the first instruments were developed for scanning densitometry using absorbance or fluorescence measurements in the reflectance or transmission mode. The commercial production of HPTLC plates in the mid-1970s provided further impetus for improvements in practice and instrumentation.