Determination of chlorhexidine in human blood by HPLC

A simple high-performance liquid chromatographic (HPLC) method for the analysis of chlorhexidine in human blood was developed. Chlorhexidine and the internal standard, chlorpheniramine, were extracted with methylene chloride under alkaline condition, and subjected to HPLC with UV detection (260 nm). HPLC separation was achieved on a polymer-based C ₁₈ column equilibrated with acetonitrile-water containing 0.05% trifluoroacetic acid, 0.05% heptafluorobutyric acid, and 0.1% triethylamine, (40:60. v/v). The calibration curve was linear in the concentration range from 1-10 μ g/ml. The concentrations of chlorhexidine in the serum and blood corpuscle of the patient who died after suspective intravenous injection of chlorhexidine gluconate were determined.