[Chiral separation of four β-blocker enantiomers using high performance liquid chromatography-tandem mass spectrometry based on amylase derivative chiral stationary phases and investigation on their separation mechanism].

A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method based on amylose derivatives as chiral stationary phases (CSPs) for the chiral separation of four β-blocker enantiomers (propanolol, metoprolol, arotinolol and carvedilol) was established. The effects of different CSPs, volume fractions of mobile phase modifiers and mobile phase additives, column temperatures and flow rates on the chiral separation of the four β-blocker enantiomers were evaluated. Baseline separation of the four β-blockers enantiomers was obtained on a Chiralpak AD-H chiral column using the mobile phase of n-hexane-ethanol-diethylamine (20:80:0.03, v/v/v) at a flow rate of 0.550 mL/min and a column temperature of 40℃. The resolutions of the enantiomers for propanolol, metoprolol, arotinolol and carvedilol were 1.37, 1.80, 2.09 and 4.70, respectively. The separation mechanism of the four β-blocker enantiomers was investigated by thermodynamic analysis and enantiomeric structure analysis. The chiral separation was all enthalpy controlled, and the cave structure of the chiral stationary phase played an important role on the separation. The method provides reference for further studies of the β-blocker enantiomers.