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By pvskumar on Tuesday, August 3, 2004 - 02:24 am:
how do we calibrate pump detector fixed loop 20 ul
0f HPLC+
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By DR on Tuesday, August 3, 2004 - 10:05 am:
In the manuals that came with the instrument, there should be procedures for verifying the performance of your components.
Generally, pump performance is verified by executing linear and step gradients using water and 0.3% acetone in water, no column. The intercept of the linear gradient's linear portion will give you a clue about the system's dwell volume.
For a fixed 20無 loop, all you can do is check precision by looking at the standard deviation (or %RSD) of replicate injections, as you chould be doing with any regular run.
Detector (I'll assume it is a UV detector) performance is measurable in terms of signal, noise, and wavelength accuracy. That said, we generally just check the reference energy at a specified wavelength adn compare the results to the manufacturer's specifications and then we do a wavelength accuracy by checking for min & max absorbances for a caffeine solution. You can also calculate absorbtivities for a series of K2Cr2O7 solutions measured at different wavelengths, but this is more time consuming and expensive. Some detectors allow you to slip in a holmium (sp?) oxide filter and check wavelength accuracy that way.
Again - RTM.
how do we calibrate pump detector fixed loop 20 ul
0f HPLC+
-------------------------------------------------------------------------------------------------------
By DR on Tuesday, August 3, 2004 - 10:05 am:
In the manuals that came with the instrument, there should be procedures for verifying the performance of your components.
Generally, pump performance is verified by executing linear and step gradients using water and 0.3% acetone in water, no column. The intercept of the linear gradient's linear portion will give you a clue about the system's dwell volume.
For a fixed 20無 loop, all you can do is check precision by looking at the standard deviation (or %RSD) of replicate injections, as you chould be doing with any regular run.
Detector (I'll assume it is a UV detector) performance is measurable in terms of signal, noise, and wavelength accuracy. That said, we generally just check the reference energy at a specified wavelength adn compare the results to the manufacturer's specifications and then we do a wavelength accuracy by checking for min & max absorbances for a caffeine solution. You can also calculate absorbtivities for a series of K2Cr2O7 solutions measured at different wavelengths, but this is more time consuming and expensive. Some detectors allow you to slip in a holmium (sp?) oxide filter and check wavelength accuracy that way.
Again - RTM.
