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At-Column Dilution for Enhancing Loading in Preparative Chromatography
Cecilia B. Mazza, Jie Cavanaugh and Tom E. Wheat
The number of samples processed and the size, or mass, of these samples define the productivity of a preparative chromatography laboratory. In addition, chromatography is the technique of choice to achieve the required purities and recoveries. Further, the chemical nature of the samples to be purified as well as the solvents they are dissolved in can vary significantly. A way that many pharmaceutical companies have found to achieve higher productivities is by increasing the level of automation. Therefore, the same system, chromatographic column and purification method may be used for a variety of samples such as the ones obtained via combinatorial chemistry methods. However, one parameter that can limit the productivity is the amount of sample that can be loaded requiring the use of a larger column or carrying out repeated runs on a small column. Both approaches consume extra time, extra solvent and increase post-purification processing. In particular, more often than not samples are dissolved in dimethylsulfoxide (DMSO) and while DMSO aids in the initial sample concentration loaded onto the column, there is the significant risk of clogging the injector and tubing leading to the column inlet due to the interaction between the sample, the solvent and the initial mobile phase conditions. Figure 1 shows the injector loop with the fluid interfaces where precipitation is likely to begin. Due to the labor and equipment costs associated with maintenance after precipitation occurs, alternative techniques are actively sought. A solution that has attracted the attention of the scientific community in the past couple of years is the At-column Dilution method.
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Waters Corporation
34 Maple Street
Milford, MA, 01757
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