What is an HPLC used for?

High-performance liquid chromatography or high-pressure liquid chromatography (HPLC) is a chromatographic method that is used to separate a mixture of compounds in analytical chemistry and biochemistry so as to identify, quantify or purify the individual components of the mixture.

What is HPLC and how it is work?

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 pressure makes the technique much faster compared to column chromatography. This allows using much smaller particles for the column packing material.

What is the types of HPLC?

Types of HPLC NP-HPLC uses polar stationary phase and non-polar mobile phase. Therefore, the stationary phase is usually silica and typical mobile phases are hexane, methylene chloride, chloroform, diethyl ether, and mixtures of these.

What is mobile phase?

The mobile phase is an inert gas that the sample is injected into that will carry it through the stationary phase, which is normally a solid. The sample solution is analyzed as it the mobile phases interacts and moves through the stationary phase.

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. Normal-phase HPLC columns have polar packing. The mobile phase is nonpolar and therefore usually an organic solvent such as hexane or methylene chloride.

What is guard column?

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.

What is run time in HPLC?

The time during a chromatographic separation is indicated as the run-time. The total time necessary for completing a chromatographic separation is slightly longer than the retention time of the last peak in the chromatogram. This time is sometimes referred to as total runtime, or length of the chromatogram.

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 are the disadvantages of HPLC?

More number of moving parts

HPLC pump operation reliability depends on clenliness of mobile phase,sample and proper operation of all four check valves during each cycle.

HPLC Pump failure can result from failure of check valves,pistons and piston seals.

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 is basic principle of HPLC?

Summary HPLC is an analytical technique used to separate, identify or quantify each component in a mixture. HPLC works following the basic principle of thin layer chromatography or column chromatography, where it has a stationary phase and a mobile phase. There are five sections in HPLC- reservoir, pump, column, detector and computer.