Stereoselective high-performance liquid chromatography determination of propranolol and 4-hydroxypropranolol in human plasma after pre-column derivatization

• Published in: Journal of chromatography. B, Biomedical sciences and applications Biomedical sciences and applications, 1997-04, Vol.692 (1), p.133-140
• Main Authors: Wu, Steven T., Ping Chang, Yu, L. Gee, Winnie, Benet, Leslie Z., Lin, Emil T.
• Format: Article
• Language: English
• Subjects: 4-Hydroxypropranolol; Adrenergic beta-Antagonists - blood; Adrenergic beta-Antagonists - pharmacokinetics; Analysis; Antihypertensive Agents - blood; Antihypertensive Agents - pharmacokinetics; Biological and medical sciences; Chromatography, High Pressure Liquid; General pharmacology; Humans; Isothiocyanates; Medical sciences; Pharmacokinetics. Pharmacogenetics. Drug-receptor interactions; Pharmacology. Drug treatments; Propranolol; Propranolol - analogs & derivatives; Propranolol - blood; Propranolol - pharmacokinetics; Reproducibility of Results; Stereoisomerism
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(96)00313-1

A stereoselective reversed-phase HPLC assay to quantify S-(−) and R-(+) enantiomers of propranolol and 4-hydroxypropranolol in human plasma was developed. The method involved liquid–liquid extraction for sample clean-up and employed 2,3,4,6-tetra- O-acetyl-β-glucopyranosyl isothiocyanate as a pre-column chiral derivatization reagent. The internal standard used was 4-methylpropranolol. The derivatized products were separated on an Altex C 18 column using a mixture of acetonitrile–water–phosphoric acid–triethylamine (58:42:0.1:0.06 and 50:50:0.15:0.06, v/v, for propranolol and 4-hydroxypropranolol, respectively) as mobile phase. The detection of propranolol derivatives was made at λ ex=280 nm and λ em=325 nm, and the corresponding 325 and 400 nm were used for 4-hydroxypropranolol derivatives. The assay was linear from 1 to 100 ng/ml and from 2 to 50 ng/ml using 0.5 ml of human plasma for propranolol and 4-hydroxypropranolol enantiomers, respectively. The present assay is used to quantify the enantiomers of propranolol and 4-hydroxypropranolol, respectively, in human plasma for pharmacokinetic studies.