Improved Ultraviolet Spectrophotometry of Serum Theophylline

We present an improved method for ultraviolet spectrophotometry of theophylline in serum. We studied various extraction techniques aimed at eliminating interferences from co-extractable serum constituents. In the resulting modified procedure, 1 ml of serum is required and a salt-solvent pair of ammonium sulfate and chloroform/hexane is used for extraction. The solvent forms the top phase after extraction, the lower phase after back-extraction, thereby permitting easy removal of the appropriate phase from culture tubes. The use of ammonium sulfate coupled with the added specificity of the extraction solvent results in an extract with low background absorption and a well-defined spectrum for the extracted theophylline.