What is the full meaning of HPLC?
The acronym HPLC, coined by the late Prof. High performance liquid chromatography is now one of the most powerful tools in analytical chemistry. It has the ability to separate, identify, and quantitate the compounds that are present in any sample that can be dissolved in a liquid.
What is an HPLC pump?
An HPLC pump is also called a “solvent delivery system.” The purpose of the pump is to maintain a constant flow of mobile phase through the HPLC system. This is accomplished regardless of the back pressure caused by the flow resistance of the HPLC column. Most HPLC pumps have a reciprocating dual piston design.
Why pump is used in HPLC?
HPLC pumps generate pressure on the solvent so it can pass through the dense column. The purpose of the pump is to force the liquid through the HPLC system while it maintains a constant flow of mobile phase. This occurs regardless of the back pressure caused by the flow resistance of the HPLC column.
How does HPLC chromatography work?
How Does HPLC 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 column packing material or adsorbent or stationary phase is typically a granular material made of solid particles such as silica or polymers.
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 …
Where is HPLC used?
HPLC has been used for manufacturing (e.g., during the production process of pharmaceutical and biological products), legal (e.g., detecting performance enhancement drugs in urine), research (e.g., separating the components of a complex biological sample, or of similar synthetic chemicals from each other), and medical …
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.
Which detectors are used in HPLC?
HPLC Detectors
- UV-Vis Detectors. The SPD-20A and SPD-20AV are general-purpose UV-Vis detectors offering an exceptional level of sensitivity and stability.
- Refractive Index Detector.
- Fluorescence Detectors.
- Evaporative Light Scattering Detector.
- Conductivity Detector.
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 the basic principle of chromatography?
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.
What is basic principle of HPLC?
Why is HPLC so important?
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 high performance liquid chromatography ( HPLC ) used for?
High Performance Liquid Chromatography (HPLC) n analytical separation technique that involves the high-pressure flow of a liquid through a column that contains the stationary phase.
How big is a sample in a HPLC?
The injectorserves to introduce the liquid sampleinto the flow stream of the mobile phase. Typical sample volumes are 5-to 20-microliters (µL). The injector must also be able to withstand the high pressures of the liquid system.
How is the flow rate of a HPLC determined?
A basic block diagram of an HPLC is shown in Figure 1. Figure 1: Block Diagram of an HPLC Your desired solvent mixture travels through capillary tubes, from the solvent reservoir to the pump, where it is becomes highly pressurized. The pump is also used to control the flow rate of the mobile phase substance,which is typically measured in mL/minute.
Which is the block diagram of a HPLC?
Block Diagram and Explanation A basic block diagram of an HPLC is shown in Figure 1. Figure 1: Block Diagram of an HPLC Your desired solvent mixture travels through capillary tubes, from the solvent reservoir to the pump, where it is becomes highly pressurized.