Enantioseparation and Determination of Propranolol in Human Plasma on a New Derivatized β-Cyclodextrin-Bonded Phase by HPLC

A 6-azido-β-cyclodextrin was synthesized and derivatized with p-nitrophenyl isocyanate as chiral ligand. Then the ligand was chemically bonded to mesoporous SBA-15 via a ‘Click Chemistry’ reaction of the azido group with alkynyl group. A new p-nitrophenylcarbamoylated β-cyclodextrin bonded SBA-15 chiral stationary phase (NPCSP) for HPLC was thus obtained. The new stationary phase was first used for enantioseparation of propranolol in human plasma under the polar organic solvent mode. The effects of methanol content, concentration of glacial acetic acid/triethylamine in mobile phase and the temperature on the enantioseparation were studied in detail. The optimized chromatographic conditions were as follows: mobile phase was acetonitrile/methanol/glacial acetic acid/triethylamine (90/10/1.25/2.25, V/V) at 288 K, with a flow rate of 0.5 mL min−1, an injection volume of 20 μL and detection wavelength at 290 nm. The resolution was 2.04 with a short run time (< 15 min) under the optimal conditions. The content of propranolol in plasma was quantitatively measured by HPLC-MS under selected ion monitoring mode ([M + H]+m/z 260.10) with hydrochlorothiazide as an internal standard. The good linear relationship was observed in the range of 2.5−50 μg L−1. The detection limits were 1 μg L−1 according to S/N = 3 for both enantiomers. The experimental results showed that the chiral stationary phase exhibited excellent chiral separation ability to propranolol. The analytical method for propranolol enantiomers was sensitive, accurate, simple and fast, which was can be for the determination of propranolol in plasma and related pharmacokinetic studies.

Graphical abstractA new p-nitrophenylcarbamoylated β-cyclodextrin bonded SBA-15 chiral stationary phase for HPLC was prepared via a ‘Click Chemistry’ reaction. The new stationary phase was first used for enantioseparation of propranolol in human plasma under the polar organic solvent mode. The content of propranolol enantiomers in plasma was quantitatively measured by HPLC-MS under selected ion monitoring mode with hydrochlorothiazide as an internal standard. The analytical method of propranolol enantiomers in plasma was sensitive, accurate, simple and fast.Figure optionsDownload full-size imageDownload as PowerPoint slide