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

• Published in: Journal of AOAC International Vol. 92; no. 3; pp. 824 - 829
• Main Authors: MOSTAFA, Gamal A. E, HEFNAWY, Mohammed M, EL-MAJED, Abdulrahman
• Format: Journal Article
• Language: English
• Published: Gaithersburg, MD AOAC International 01.05.2009; Oxford University Press
• Subjects: Biological and medical sciences; Chromatography, High Pressure Liquid - methods; Clenbuterol; Clenbuterol - blood; Clenbuterol - chemistry; Drug Stability; Food industries; Fundamental and applied biological sciences. Psychology; High performance liquid chromatography; Humans; Identification and classification; Measurement; Methods; Pharmaceutical research; Stereoisomerism; Vancomycin - chemistry; United States; HPLC chromatography; Chiral stationary phase; Antibiotic; Clenbuterol; Separation; Vancomycin
• ISSN: 1060-3271; 1944-7922
• DOI: 10.1093/jaoac/92.3.824

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 methanol-triethylamine-glacial 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 0.2-20 microg/mL, and an average recovery of 98.0% and a mean relative standard deviation of 1.5% were obtained at 5.0 microg/mL. The lower limit of detection was 0.05 microg/mL for each enantiomer. The mean recovery for R-(-) and S-(+)-clenbuterol enantiomers from plasma was 91.0-97.0% at 0.20-20 microg/mL. The method was successfully used to identify and quantify the clenbuterol enantiomers in human plasma.