Purification

  • Chromatographic result of the purification of Tryptamine. Column: Kromasil EternityXT 10 µm C18, 4.6 x 250 mm, elution at pH 2.5 and pH 11, respectively, with acetonitrile / 0.1 M potassium phosphate buffer [20/80], temperature 22 °C, flow rate 1.0 mL/min, detection: UV @ 280 nm.

Fine tuning productivity and yield in purification with the use of pH

Dec 13 2017 Read 670 Times

The issue of loadability under certain chromatographic conditions is critical when drug companies need to isolate material for characterization, toxicity studies and also manufacturing. The amount of sample that can be loaded onto the column to obtain the key substance at a required purity, translates into a productivity and yield, which in turn will influence costs and determining the feasibility of a project.

The example shown in the figure, illustrates two preparative chromatograms for the purification of alkaloid tryptamine, from an impurity that elutes before the compound of interest. On the left hand side, the chromatographic result shows the sample is loaded onto the column equilibrated at pH of 2.5. Here the impurity elutes under the main amine peak affecting productivity and yield. On the other hand, the chromatographic result shown on the right hand side indicates that as amines are better retained under basic conditions, it is possible to separate the impurity that elutes ahead of the main peak and therefore obtain better purification results. These results exemplify the benefits of exploring pH as an option when carrying out preparative chromatography and choosing a stationary phase material that can withstand a wide range of pH.

Designed to address analytical and preparative chromatography under low, neutral and high pH, in combination with also aggressive inorganic buffers and/or elevated temperatures, Kromasil EternityXT UHPLC and HPLC family of columns are provided in a wide range of particle sizes, between 1.8 µm and 10 µm, and analytical to preparative dimensions for fast turnaround, easy method transfer and seamless scale-up from the discovery stage to manufacturing.

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