High-performance liquid chromatography (sometimes referred to as high-pressure liquid chromatography), HPLC, is a chromatographic technique used to separate a mixture of compounds in analytical chemistry and biochemistry with the purpose of identifying, quantifying or purifying the individual components of the mixture. HPLC is considered an instrumental technique of analytical chemistry (as opposed to a gravitimetric technique). HPLC has many uses including medical (e.g. detecting vitamin D levels in blood
UHPLC+ Focused: Ultra High Performance Liquid Chromatograph
High-performance liquid chromatography (sometimes referred to as high-pressure liquid chromatography), HPLC, is a chromatographic technique used to separate a mixture of compounds in analytical chemistry and biochemistry with the purpose of identifying, quantifying or purifying the individual components of the mixture. HPLC is considered an instrumental technique of analytical chemistry (as opposed to a gravitimetric technique). HPLC has many uses including medical (e.g. detecting vitamin D levels in blood serum), 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 manufacturing (e.g. during the production process of pharmaceutical and biological products).
Chromatography can be described as a mass transfer process involving adsorption. HPLC relies on pumps to pass a pressurized liquid and a sample mixture through a column filled with a sorbent, leading to the separation of the sample components. The active component of the column, the sorbent, is typically a granular material made of solid particles (e.g. silica, polymers, etc.), 2-50 micrometers in size. The components of the sample mixture are separated from each other due to their different degrees of interaction with the sorbent particles. The pressurized liquid is typically a mixture of solvents (e.g. water, acetonitrile and/or methanol) and is referred to as "mobile phase". Its composition and temperature plays a major role in the separation process by influencing the interactions taking place between sample components and sorbent. These interactions are physical in nature, such as hydrophobic (dispersive), dipole-dipole and ionic, most often a combination thereof.
HPLC is distinguished from traditional ("low pressure") liquid chromatography because operational pressures are significantly higher (50-350 bar), while ordinary liquid chromatography typically relies on the force of gravity to pass the mobile phase through the column. Due to the small sample amount separated in analytical HPLC typical column dimensions are 2.1 - 4.6 mm diameter, and 30 - 250 mm length. Also HPLC columns are made with smaller sorbent particles (2- 5 micrometer in average particle size). This gives HPLC superior resolving power when separating mixtures, which is why it is a popular chromatographic technique.
The schematic of an HPLC instrument typically includes a sampler, pumps, and a detector. The sampler brings the sample mixture into the mobile phase stream which carries it into the column. The pumps deliver the desired flow and composition of the mobile phase through the column. The detector generates a signal proportional to the amount of sample component emerging from the column, hence allowing for quantitative analysis of the sample components. A digital microprocessor and user software control the HPLC instrument and provide data analysis. Some models of mechanical pumps in a HPLC instrument can mix multiple solvents together in ratios changing in time, generating a composition gradient in the mobile phase. Various detectors are in common use, such as UV/Vis, photodiode array (PDA) or based on mass-spectrometry. Most HPLC instruments also have a column oven that allows for adjusting the temperature the separation is performed at.
UHPLC+ Focused: UHPLC for All Standard LC Systems
UHPLC delivers important benefits—faster runs, better resolution, and lower operating costs. Thermo Fisher now offers UHPLC performance at standard HPLC prices.
UHPLC compatibility has been added to all of the standard and basic automated HPLC systems and extended the UHPLC performance of the UltiMate® 3000 RSLC systems, giving you more capability for your analytical needs.
The UltiMate 3000 HPLC system offers UHPLC compatibility across all modules, ensuring maximum performance. Covering flows from 20 nL/min to 10 mL/min and offering a wide range of pumping, sampling, and detection modules, the UltiMate 3000 series provides solutions for all your chromatography needs.
Features and Benefits
- UHPLC design for all nano, standard analytical, and RSLC systems
- 620 bar maximum pressure for basic and standard analytical systems—setting a new benchmark in HPLC
- x2 Dual Systems form a unique platform for routine analysis, increased productivity solutions, and advanced chromatographic techniques
- Systems are controlled using Chromeleon® Chromatography Data System software—providing Intelligent Functionality and Operational Simplicity™
- Systems are offered with Viper™ and nanoViper™ fitting systems—fingertight, zero-dead-volume connections even at UHPLC pressures
· UHPLC delivers major benefits, we all know this – faster runs, better resolution, and lower running costs. Yet migrating methods to UHPLC cannot always be justified because of the investment step in UHPLC technology.
· This is no longer true.
Thermo Scientific now offers UHPLC performance at standard HPLC costs.
· Thermo Scientific has added UHPLC compatibility to all of its standard and basic automated HPLC systems, and extended the UHPLC performance of our Thermo Scientific Dionex UltiMate® 3000 RS systems – giving you more opportunities to get the answers you are looking for, today and tomorrow. + Industry leading Chromeleon® software + Corona® Charged Aerosol Detectors + Applications D-library. Never before has so much power been available in a single package and for such a low initial investment.
· Now everyone can have the confidence to move to UHPLC technology.
· Thermo Scientific – the only HPLC brand uniquely focused on making UHPLC technology available to all users, all laboratories, and for all analytes.