Problems affecting the liquid chromatographic quantitation of chlorhexidine digluconate in ophthalmic solutions.

A liquid chromatogrpahic (LC) assay was recommended by the Food and Drug Administration of the United States for the uptake and release study of chlorhexidine digluconate (CHDG) in ophthalmic solutions by contact lenses. The results from this and other reversed-phase LC assays of CHDG were inaccurate when the working standard and sample solution matrices were different. The error was caused by binding of the analyte onto the container surface and LC column packings. The loss of chlorhexidine due to binding was dependent upon, and very sensitive to, the counter ions in the sample solutions. Relative to water solutions of CHDG, solutions containing chloride, thiosulphate and edetate reduced the loss of the analyte, while solutions containing borate enhanced the loss. To assay CHDG reliably with reversed-phase LC, the media of the working standard and the sample solutions should be closely matched.