Detection of beta-blockers in urine by solid-phase extraction-supercritical fluid extraction and gas chromatography-mass spectrometry.

The most convenient way to perform supercritical fluid extraction (SFE) of liquid sample matrices is to combine it with solid-phase extraction (SPE). beta-Blockers from urine were collected on an Empore disc, which was then placed into an extraction cell for derivatization and SFE. SPE recovery was best at pH 10. Effects of temperature, pressure and volume of pyridine on the acetylation and SFE processes were studied. Without acetylation the beta-blockers were not significantly soluble in CO2. SFE temperatures of 70 degree C and 150 degree C together with 200 microliters of acetic anhydride and 400 microliters pyridine gave the best results. With the SPE-SFE-GC-MS method developed here, beta-blockers like oxprenolol, metoprolol and propranolol could easily be detected in urine samples, and the limit of detection (LOD) for these compounds was found to be 20 ng/ml, 30 ng/ml and 40 ng/ml, respectively.