What is the instrumentation of HPLC?
What is the instrumentation of HPLC?
HPLC instrumentation is made up typically of nine basic components: mobile phase/solvent reservoir, solvent delivery system, sample introduction device, column, post-column apparatus, detector, data collection and output system, post-detector eluent processing, and connective tubing and fittings.
What are the 5 parts of HPLC and their function?
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 is the basic principle of HPLC?
The separation principle of HPLC is based on the distribution of the analyte (sample) between a mobile phase (eluent) and a stationary phase (packing material of the column). Depending on the chemical structure of the analyte, the molecules are retarded while passing the stationary phase.
Is HPLC an instrument or equipment?
HPLC (known as high performance liquid chromatography, high pressure liquid chromatography) is used to separate the phases of a solid or liquid analyte, regardless of its stability and volatility. HPLC equipment is popular for its regarded ease of set up, use, and configuration potential.
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.
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.
How many pumps are in HPLC?
There are 3 main types of HPLC pumps: 1. reciprocating pump – 90% usage. The solvent is drawn into a small chamber (when the solvent check valve is open) and pumped out of it (when the column check valve is open) by the back and forth motion of a motor-driven piston (Fig 28-6 or R5 Fig 6.9).
How many detectors are there in HPLC?
They are of three types, i.e. fixed wavelength detectors, variable wavelength detectors and the diode array detectors.
Why high pressure is used in HPLC?
Smaller particle sizes [<10 microns] are required to improve separation power. However, smaller particles have greater resistance to flow, so higher pressures are needed to create the desired solvent flow rate. Pumps and columns designed to withstand high pressure are necessary.
Why is 254 nm used in HPLC?
254 nm is widely used for HPLC-UV detection mainly for historical reasons, since the mercury lamps that were used in early HPLC detectors emit the brightest light at 254 nm. Thus, detection at that wavelength would have provided the greatest sensitivity. Some early UV detectors were “fixed” at that wavelength.