Determination of 2,2,2-trichloroethanol in plasma and urine by ion-exclusion chromatography
Abstract
A simple and rapid chromatographic method has been developed for the determination of 2,2,2-trichloroethanol (TCEOH) and its glucuronide in plasma and urine. A glass column (150 × 6.6 mm i.d.) packed with Aminex A-5 cation-exchange resin (potassium form) following the slurry method was used as the analytical column, and an admixture of 10 mmol l–1 potassium sulfate and 10 mmol l–1 potassium hydroxide solution as the eluent (pH 12.2). Diluted plasma samples and urine samples were directly injected into the chromatograph through a 0.45 µm membrane filter without deproteinization. The amount of TCEOH conjugated to glucuronide was determined following treatment with β-glucuronidase (200 U) for 30 min at 37 °C. This allowed the concentration of free, total, and conjugated TCEOH to be determined. The calibration graph was rectilinear from 5 to 500 mg l–1 of TCEOH, with a detection limit of 3 mg l–1, 2σ, being the signal-to-noise ratio. The analytical recovery of TCEOH, obtained by analysing spiked plasma and urine samples, was in the range 98.4–102% and the relative standard deviation was less than 3.5%