This Application Note demonstrates the mass-based purification of low gram amounts of compounds using the Agilent 1100 Series purification platform. Starting the experiments on 21.2-m i.d. columns two application examples were scaled up to a 50-mm i.d. column operated at 100 mL/min. Fractions with purities of 85 and 95 % were obtained by mass-based fraction collection even though the column overload did not allow the UV-trace to show separated peaks.
The document describes a method to purify up to 4 grams of crude product in a single run on 50 mm diameter columns operated at 100 mL/min. It utilizes mass-based fraction collection mode, which allows for the isolation of compounds with high purity even when UV-trace peaks are not separated due to column overload.
The text details the specific equipment configuration required, including preparative pumps, a dual-loop autosampler, and a multiwavelength detector. Two test mixtures containing calcium channel blockers were used to validate the method on both analytical-scale (21.2 mm i.d.) and preparative-scale (50 mm i.d.) columns.
The system can purify up to 4 grams of a crude product in a single run on 50 mm diameter columns.
The experiments were operated at a flow rate of 100 mL/min.
The re-analysis of the collected fraction presented a purity of 96.2% for nimodipin.