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Each component in the sample interacts differently with the adsorbent material, causing different migration rates for each component. [3] [better source needed] These different rates lead to separation as the species flow out of the column into a specific detector such as UV detectors. The output of the detector is a graph, called a chromatogram.
Liquid chromatography as we know it today really got its start in 1969, when the first modern HPLC was designed and marketed as a nucleic acid analyzer. [9] Columns throughout the 1970s were unreliable, pump flow rates were inconsistent, and many biologically active compounds escaped detection by UV and fluorescence detectors. Focus on ...
In liquid chromatography, the mobile phase velocity is taken as the exit velocity, that is, the ratio of the flow rate in ml/second to the cross-sectional area of the ‘column-exit flow path.’ For a packed column, the cross-sectional area of the column exit flow path is usually taken as 0.6 times the cross-sectional area of the column.
Another advantage to using APCI over other ionization methods is that it allows for the high flow rates typical of standard bore HPLC (0.2–2.0 mL/min) to be used directly, often without diverting the larger fraction of volume to waste. Additionally, APCI can often be performed in a modified ESI source. [25]
The charged aerosol detector (CAD) is a detector used in conjunction with high-performance liquid chromatography (HPLC) and ultra high-performance liquid chromatography (UHPLC) to measure the amount of chemicals in a sample by creating charged aerosol particles which are detected using an electrometer.
As can be seen in Figure 1, these detectors have a light source, a dispersion element that is a diffraction grating or prism, a flow cell, to where the sample arrives directly from the chromatographic column, an optical bench of lenses and mirrors, and a diode that receives the light coming from the optical system and translates it into a ...
Fast protein liquid chromatography (FPLC) is a form of liquid chromatography that is often used to analyze or purify mixtures of proteins. As in other forms of chromatography, separation is possible because the different components of a mixture have different affinities for two materials, a moving fluid (the mobile phase) and a porous solid (the stationary phase).
Regarding applicability to a range of HPLC flow rates, the signal level of analytes by APPI has been observed to saturate and even decay at higher solvent flow rates (above 200 μl/min), and therefore, much lower flow rates are recommended for APPI than for ESI and APCI.
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