, Volume 24, Issue 1, pp 155-164

Direct injection of plasma into column liquid chromatographic systems

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Summary

Since the late 1970’s several techniques involving the direct injection of biological fluids, like plasma and serum have been published. Most of the systems presented utilize a precolumn for the reception of the biological fluids, trace enrichment and preliminary clean-up before the analyte fraction is transferred to the separation column. The most common solid phase for the precolumn is silanized silica, but many other types have been used. One was especially designed for the purpose of direct injections of biological fluids: the Internal Surface Reversed Phase support.

The main problem with direct injections is the development of column instabilities due to the denaturation and precipitation of plasma proteins on the support. It has, however, been possible to design systems which can handle the injection of 25ml total plasma volume without the need to exchange precolumn. The most promising techniques in this respect are the so called “pre-column” venting plug technique and the use of micellar mobile phases.

Losses of analytes may be a problem when protein-binding extends 95%. However, remedies are available, such as the use of more hydrophobic solid phases, the addition of competitive binders and pH adjustments of injected solutions.