| Microcolumn High Performance Liquid Chromatography (micro-HPLC) is one of the miniaturized cbromatographic separation techniques that have recently been under extensive development .The main advantages due to miniaturization are higher efficiency, lower consumption of sample, mobile and stationary phase and less environmental pollution. In this paper, micro-HPLC instrumentation compatible with 0.5-1 .OmmI.D. packed capillary HPLC columns was set up using standard commercial HPLC system. All experiments were carried out on this mirco-HPLC system. A zero-dead volume column end and a conical shaped slurry reservoir for wide-bore stainless-steel packed capillary HPLC columns were designed and evaluated. A detailed procedure for the preparation of wide-bore stainless-steel packed capillary HPLC columns with 0.5-1.0mm I.D. was described. The influence of packing direction, packing solvent, slurry solvent, slurry concentration, packing pressure, column length and the packing material on the column performance has been studied systematically. Columns were evaluated by the reduced plate height vs. reduced velocity curve and the peak asymmetric factor. Experimental results showed that the column efficiency and the reproducibility were better than 70% of theoretical value and 6%RSD, respectively. Separations of different samples were demonstrated. Abstract Gradient elution system for use with wide-bore stainless-steel packed capillary HPLC columns was described as well as the repeatability of the system. Replicate chromatograms obtained from wide-bore stainless-steel packed capillary HPLC columns under gradient-elution development had been shown to have good retention time repeatability and precision.
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