High Performance Liquid Chromatography
High Performance Liquid Chromatography (HPLC) is a form of liquid chromatography to separate compounds that are dissolved in solution. HPLC instruments consist of a reservoir of mobile phase, a pump, an injector, a separation column, and a detector. Compounds are separated by injecting a plug of the sample mixture onto the column. The different components in the mixture pass through the column at different rates due to differences in their partitioning behavior between the mobile liquid phase and the stationary phase. Instrumentation Solvents must be degassed to eliminate formation of bubbles.
The pumps provide a steady high pressure with no pulsating, and can be programmed to vary the composition of the solvent during the course of the separation. The liquid sample is intro- duced into a sample loop of an injector with a syringe. When the loop is filled, the injector can be inject the sample into the stream by placing the sample loop in line with the mobile phase tubing.
The different types of HPLC columns are described in a separate document. The presence of analytes in the column effluent is recorded by detecting a change in refractive index, UV-VIS absorption at a set wavelength, fluorescence after excitation with a suitable wavelength, or electrochemical response. Mass spectrometers can also be interfaced with liquid chromatography to provide structural information and help identify the separated analytes.
SYKAM S 1122 HPLC Pump
The HPLC pump S 1122 is a dual plunger solvent delivery system. The
mechanical design includes short plunger stroke technology with only two check valves. This results in low pulsation, typically observed with dual plunger pumps and high reliability otherwise achieved only by single plunger pumps. The delivery plunger performs a two mm stroke, the compensation plunger makes one mm. The nearly pulseless solvent delivery of the pump results from the high stroke frequency and the use of the compensation plunger. The two plungers are counter rotating installed. While the delivery plunger pushes out the solvent volume, the compensation plunger, located on the pressure side, collects half of the volume. During the return movement of the delivery piston, the collected volume is pumped from the compensation plunger.
The microprocessor controlled speed regulation leads to a very stable speed of the D.C. motor, driving the plungers. Additionally, an integrated cal- culation software continuously corrects the pressure dependent differential between the preset and actual flow rates. This results in a very constant volume delivery throughout the entire range, independently of the resulting back pressure.
S 2100 Solvent Delivery System
The HPLC Pump S 2100 is a compact eluent dosing system, upgradable from a high performance isocratic pump to a quaternary gradient pump.
- menu programmable through large graphical display
- basic system: isocratic pump with three interchangable pump heads
- materials for the pump head: standard stainless steel, optional PEEK, Titan or PVDF
- Quaternary gradient operation
S 5200 AUTOSAMPLER
The system can be upgraded at any time from the basic version for fixed volume injection with sample loop overfilling, up to variable volume injection mode for variable volumes from 1 - 100 µl (optional up to 500µl) without changing the sample loop.
With the next upgrading step, the S 5200 can be used for fully automatic sample processing fro pre-column derivatization. In the derivatisating mode, up to three independent reagents can be added to a specified sample volume, mixed with sample and programmed with independent reaction times. In addition, the sample can be diluted with more than 100 times (depending on sample volume and syringe volume).
For temperature sensitive samples, the sample tray can be cooled or heated with the optional cooling system.
For chemical inert operation, the instrument can be delivered for fully
metal-free sample handling (completely equipped with PEEK and FEP materials, including sample needle).
A wide range of sample racks and sample vials can be used for operation from 1 µl up to 5000 µl or by using micro titer plates. Simple operation through large alphanumeric display menu.
Sykam offers the full range of HPLC detectors from routine UV/Vis Detectors and RI Detectors to sophisticvated Electrochemical, Conductivity and Fluorescence Detectors. With this range Sykam can offer a solution for any application.
S 3240 UV/VIS DETECTOR
Diode Array Technology, Photoconductor Optics and Dual Lamp Source.
- Diode Array Technology for wavelength change without any mechanical moving parts.
- 4-Channel measurement with separated analog output (0 - 1000 mV)
- Online scan of wavelength and collection of spec- tra data without stopping the flow or interrupting the running analysis.
- Dual lamp (deuterium/tungsten) for a spectral range of 190 - 720 nm.
- Integrated peak detector with programmable delay time.
- Integrated soolvent recycling system (3-way sole- noid valve optionally).
- External START, external AUTOZERO via TTL- signal.
- Serial interface (RS232C) for external control.
- Programmable change of wavelength (up to 10 steps).
- Full DAD operation mode with ChromStar software (optional).
S 3580 RI DETECTOR
The S 3580 Differential Refractive Index Detector series offers the sensitivity, stability and reproduce- bility required for optimal RI detection. The thermal isolated optic with a countercurrent heat exchanger and with its programmable temperature control, results in an extremly stable baseline and an optimal Signal / Noise ratio.
The S 3580 series provide autopurge and autozero capabilities, as well as RS232 communication to acquire data directly without using any external sig- nal interface. S 3580 detectors are available for:
- semi-preparative mode
Degasser & Reagent Organizer
The Sykam Reagent Organizers, with integrated gas supply and valve-equipped bottles provide a clean solution for storing solvents on your Chromatograph.
The integrated gas supply can be used for degassing or for keeping solvents oxygen-free. Additionally, Sykam offers a Vacuum Degasser for keeping your solvents gas free to insure a proper working of the Chromatograph.
S 7505/7510 VACUUM DEGASSER
The degassing of solvents is very important in liquid chromatography, as oxygen and other in the sol- vent dissolved gases affect the analysis for various reasons; through the volume contraction (gradient mixing), different pressure ratio (in front and behind the column) and caused by different temperatures, the dissolved gases are released in course of time. This will influence the reliability of the analysis (pump) as well as the detection efficiency (measuring cell). Therefore, dissolved gases have to be removed from the solvent.
S 7131 REAGENT ORGANIZER
The S 7131 Reagent Organizer is the optimal solution for storing your solvents. The integrated gas sup- ply with pressure regulator let you store the eluent under inert-gas pressure, or for degassing purposes using Helium.
The organizer can hold up to four eluent bottles, which can be closed individually with the integrated valve bottle caps. The S 7131 Reagent Organizer could be used with the S 2100 Solvent Delivery System.
For data acquisition, integration and instrument control, Sykam offers software packages at different price and feature levels to meet the individual customers needs.