By Donald Parriott
This advisor for the training chromatographer who wishes a prepared resource of data on HPLC detection explores and compares latest detection structures and detectors, outlines the typical difficulties linked to a given detector, and gives confirmed techniques to warding off such difficulties.
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Extra resources for A Practical Guide to HPLC Detection
Some manufac turers use a simple R/C (resistor/capacitor) circuit on the detector output, while other vendors use more complex circuits, such as a second-order Bessel filter. To cloud the waters even more, two different definitions for this function are commonly used: Suppose that a slug of sample is injected with a sampling valve into a tube connected directly to a detector, with an LC p u m p supplying mobile phase to the injection valve. Assuming plug flow, when the plug of sample reaches the detector, an instantaneous change in concentration would occur, causing a change in the detector output.
Mobile phase from the HPLC column entering the flow cell is often at a different temperature than the flow cell compartment, especially if a column oven is used. As the mobile phase enters the flow cell, the resultant variations in 57 3 · Absorbance Detection Mean Response Factor R E S P O N S E F A C T o 95 % of Mean Response Factor Mean Response Factor Peak Height Curve 7 o R Peak Height R E S P Peak Area Curve N S E F A C T o R 95% of Mean Response Factor Peak Area —i— 10 100 1000 10,000 Concentration FIG.
7 times the column-bed volume. The column-bed volume is given by the volume of a cylinder. 23 cm. 66 mL Also, k' is by definition the number of column void volumes past the void volume (retention volume of an unretained peak) in which a peak elutes. 6mL)(2ff) 1/2 You also know something about your hardware. You have a good variable wavelength detector with noise of only 2 x 10 ~5 absorbance units at the ab sorbance maximum of your drug, and with a cell path length of 1 cm. Several detectors on the market can meet this specification.
A Practical Guide to HPLC Detection by Donald Parriott