Optimization of a column liquid chromatographic procedure for the determination of plasma salbutamol concentration

A reversed-phase column liquid chromatographic procedure with fluorescence detection for the determination of salbutamol in plasma is described. A l-ml aliquot of the sample, after the addition of bamethan as the internal standard, is passed through a Bond Elut silica extraction column. The column is selectively washed to remove neutral, acidic, and weakly basic compounds. The desired compounds are eluted with a l-ml aliquot of methanol. The eluate is evaporated under vacuum at ambient temperature and the residue is reconstituted in 40 microliters of the mobile phase which contains octanesulfonic acid as the ion-pairing reagent. The entire extract is injected onto a 150 x 4.6 mm I.D. column packed with 5-micron octylsilica particles. Peaks are detected with a fluorescence detector (excitation wavelength = 275 nm, emission wavelength = 310 nm). In the resulting chromatogram, salbutamol and the internal standard give sharp peaks that are well resolved from the extraneous peaks. The procedure allows the quantitation of salbutamol down to 0.2 ng/ml.