What is the function of HPLC?
The main purpose of the HPLC technique is to identify, quantify and purify a particular analyte or compound. Both quantitative and qualitative analysis can be done.
What is the principle of liquid solid chromatography?
Liquid-solid chromatography utilizes a solid stationary phase, and the major mechanism of retention is adsorption. Popular adsorbents are silica and alumina, which both retain polar compounds. If a polar mobile phase is used, the solutes are rapidly swept from the bed.
What is HPLC and its applications?
High performance liquid chromatography or commonly known as HPLC is an analytical technique used to separate, identify or quantify each component in a mixture. The mixture is separated using the basic principle of column chromatography and then identified and quantified by spectroscopy.
What is HPLC and its types?
Different types of HPLC columns are used in analysis of different pharmaceutical compounds according to their nature and column separation capacity. Therefore silica columns can be used to analyze the compounds of different chemical natures. The material filled in the HPLC columns is known as a stationary phase.
Which solvent is used in HPLC?
The A solvent is generally HPLC grade water with 0.1\% acid. The B solvent is generally an HPLC grade organic solvent such as acetonitrile or methanol with 0.1\% acid. The acid is used to the improve the chromatographic peak shape and to provide a source of protons in reverse phase LC/MS.
What are the parameters of HPLC?
The HPLC parameters are the: efficiency factor(N), the retention factor (kappa prime), and the separation factor (alpha). Together the factors are variables in a resolution equation, which describes how well two components’ peaks separated or overlapped each other.
What is chromatography principle?
Chromatography is based on the principle where molecules in mixture applied onto the surface or into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase. Based on this approach three components form the basis of the chromatography technique.
Which pump is not used in HPLC?
Which of the following is not an advantage of Syringe type pumps used in High-pressure liquid chromatography? Explanation: The limitation of Syringe type pump is that it has the limited solvent capacity and is inconvenient when solvents are to be changed.
What are the detectors used in HPLC?
Lesson 6: Detectors for HPLC
- UV, VIS, and PDA Detectors. Refractive-Index Detector.
- UV, VIS, and PDA Detectors.
- Refractive-Index Detector.
- Evaporative Light Scattering Detector.
- Multi-Angle Light Scattering Detector.
- Mass Spectrometer.
- Conductivity Detector.
- Fluorescence Detector.
What is guard column in HPLC?
A guard column is a protective column or cartridge installed between the injector and the analytical column. It serves to remove the impurities and suspended solids from reaching the analytical column. Typically it has a length of about 2 cm and internal diameter of 4.6 mm.
Why C18 column is used in HPLC?
C18 is considered better for separating compounds such as long-chain fatty acids as compared to relatively small organic compounds. It is relatively cheap as it is produced in large amounts by most manufacturers.
What is the principle that is used in HPLC?
The principle involved in HPLC testing is the separation of compounds in a mixture more efficiently and also quickly than that of traditional column chromatography. Segregation of compounds is due to their relative differences in travel through the column on the application of pressure exerted through the mobile phase or carrying liquid.
What are the disadvantages of HPLC?
More number of moving parts
What is HPLC and how does it work?
HPLC is a technique for separation, identification and quantification of components in a mixture. It is especially suitable for compounds which are not easily volatalised, thermally unstable and have high molecular weights. The liquid phase is pumped at a constant rate to the column packed with the stationary phase.
What is difference between HPLC and HPTLC?
The key difference between HPLC and HPTLC is that HPLC allows quantitative separation of components in a sample whereas HPTLC does not allow quantitative separation of components in a sample. Chromatography is a physical separation method that is used to separate and identify the components in a mixture.