A Practical Guide to HPLC Detection by Donald Parriott

By Donald Parriott

This advisor for the training chromatographer who desires a prepared resource of knowledge on HPLC detection explores and compares current detection structures and detectors, outlines the typical difficulties linked to a given detector, and gives confirmed methods to fending off such difficulties.

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Bonded phase column 10 20 30 minutes FIG. 2°C change in column temperature. Top baseline for an inert glass bead column and bottom baseline for an NH 2 bonded phase column. 0 mL/minute flow rate. The column was equilibrated at ambient temperature, and, at the time indicated by "heater on," or time zero, the column was transferred to the heater block. At the time indicated by "heater off" the column was removed from the heater block and equilibrated with the ambient air temperature. Soon after the column was transferred to the heater block, there was a positive excursion in the detector baseline, which returned almost to the original baseline position before the column was removed from the heater block.

However, the back pressure produced by the tubing is proportional to the flow rate, the tubing length, and inversely proportional to the fourth power of the tubing diameter. ) connect­ ing tubing is used, place the detector as close to the column as possible and use a short length of connecting tubing. d. d. columns. d. Teflon tubing (Beckman), with the same tube fittings, is excellent. d. d. d. d. d. d. d. PEEK tubing from Upchurch for connecting the column outlet to the detector. There is a low dispersion connection trick for old detectors, such as the Altex 153, which use Cheminert type fittings in the flow cell.

D. column. 001 AUFS. 5 μΐ. 5 ^L flow cell without heat exchanger. 3 · Absorbance Detection ■WfMfr^jM ^vw** J*^ 5 Time (min) 0 Time (min) Jj| FIG. d. column. 001 AUFS. 5 μΐ. flow cell without heat exchanger. Roger Blain 60 J^jvi^ 0 5 0 5 Time (min) Time (min) JJ1 u iff Time (min) Time (min) 0 5 Time (min) I » YUtl4iftitiiA/ Time (min) FIG. d. column. 001 AUFS. 5 ^L flow cell without heat exchanger. 3 · Absorbance Detection 61 3. Some Practical Hints The tubing used to connect the column to the detector is also a source of extra-column band broadening.

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