A novel assay for the distribution of pyrithione biocides in bacterial cells.

Sodium pyrithione and zinc pyrithione (NaPT and ZnPT, respectively) are widely used as cosmetic preservatives and metal chelating agents. They are commonly assayed using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). However, a simple quantitative colorimetric assay has not been previously reported for these compounds. This paper describes the development of a spectrophotometric assay for the quantification of the pyrithiones which is based on the chelation of copper (II) ions by the biocides. This assay was developed in order to facilitate the determination of the distribution of these biocides in the Gram-negative bacteria Escherichia coli NCIMB 10,000 and Pseudomonas aeruginosa NCIMB 10,548. Sodium pyrithione was exhibited only in the cytosol of E. coli and Ps. aeruginosa. Zinc pyrithione, however, was assayed in the cytosol of both bacteria and was found in the cell envelope of Ps. aeruginosa. These findings suggest that the pyrithione biocides are active within bacterial cells as well as at the cell membrane.