By Donald Parriott
This advisor for the training chromatographer who wishes a prepared resource of data on HPLC detection explores and compares current detection structures and detectors, outlines the typical difficulties linked to a given detector, and provides confirmed methods to warding off such difficulties.
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Additional info for A Practical Guide to HPLC Detection
This works only if you don't have a "busy" chromatogram with lots of extra peaks that would interfere with the requisite autozero after a wavelength change. Short lamp life, at around 350 to 500 hours, used to be a major problem with these detectors, but the new Westinghouse lamps are good for more than 1000 hours. This still doesn't equal the 2000 plus hours of lamp life claimed for some mercury lamps. Variable wavelength detectors are more mechanically complex, and hence more expensive and less reliable than fixed wavelength detectors.
Waters Associates pioneered the use of a tapered flow cell with the Model LAMP FILTER FLOW CELL BODY PHOTODIODES APERTURE FIG. 3-7 Dual path flow cell. 47 3 · Absorbance Detection REFRACTIVE INDEX n FIG. 3-8 Snell's law: r^ sin a = n2 sin Θ. 440 detector. The idea is that the flow cell diameter diverges toward the outlet window at a wider angle than the light is bent when it enters the mobile phase (Fig. 3-9). Thus the light never has a chance to strike the wall of the cell. Detectors using this cell design are relatively unaffected by changes in refractive index.
3-7). b. Compensation for refractive index changes Another design consider ation is compensation for refractive index changes. Solutions with different refractive indices bend light to differing degrees, according to Snell's law (Fig. 3-8). When the refractive index of the mobile phase changes, either because of 46 Roger Blain hv INCIDENT LIGHT hv PARTIALLY SILVERED MIRROR FIG. 3-6 Beam splitter. gradient elution or due to a sample solvent different than the mobile phase eluting at the void volume of the column, a shift in the baseline occurs.