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A practical procedure is described for the modification of a conventional HPLC system to a set-up compatible with high speed HPLC requirements. A number of experimental parameters, such as extra-column volume, extra-column band broadening, effectiveness, and dwell volume, were examined to characterize and demonstrate the suitability of the modified system for use in high speed HPLC applications. In addition, an injector bypass is presented as an alternative to reduce the pressure damage suffered by short columns packed with microparticles.

A fast HPLC method has been developed for simultaneous determination of sultamicillin and its synthesis precursors. The analytes are separated in 2.5 min by means of a Kromasil 100 C18 column (50 mm  2.1 mm i.d., 3.5 mm) at 25 C. The mobile phase (A: 5 mM KH 2 PO 4 and 20 mM KCl adjusted to pH 6.0 with H3PO4 plus 1% THF and B: acetonitrile with 1% THF) was pumped at a flow rate of 0.5 mL min À1 according to the fast gradient mode: 0-0.9 min, 40% B; 0.9-1.0 min, 85% B; 1.0-2.5 min, 85% B; 2.5-2.6 min, 40% B, 2.6-4.0 min, 40% B. Detection was by ultraviolet absorbance at 205 nm. The method was validated in accordance with the International Conference on Harmonisation (ICH) guidelines, good accuracy, intermediate precision ( 3.8%), and linearity being observed for all compounds. This method is sensitive (limits of detection ranged between 0.1-1.1 mg l À1 ) and selective for quantifying sultamicillin and its synthesis precursors and could be used for in-process control.