Separation and Determination of Clenbuterol by HPLC Using a Vancomycin Chiral Stationary Phase

Abstract

Abstract Enantiomers of clenbuterol were separated by a new HPLC method on a chiral column. Enantiomeric resolution was achieved on a vancomycyin macrocyclic antibiotic chiral stationary phase known as chirobiotic V with UV detection at 247 nm. The polar ionic mobile phase consisting of methanoltriethylamineglacial acetic acid (100 + 0.05 + 0.025, v/v/v), was used at a flow rate of 1.0 mL/min. The method was validated for linearity, accuracy, precision, and robustness. Standard linear calibration curves were established for the R-() and S-(+) enantiomers over the range of 0.220 g/mL, and an average recovery of 98.0 and a mean relative standard deviation of 1.5 were obtained at 5.0 g/mL. The lower limit of detection was 0.05 g/mL for each enantiomer. The mean recovery for R-() and S-(+)-clenbuterol enantiomers from plasma was 91.097.0 at 0.2020 g/mL. The method was successfully used to identify and quantify the clenbuterol enantiomers in human plasma.