HPLC column-switching technique for sample preparation and fluorescence determination of propranolol in urine using fused-core columns in both dimensions

• Published in: Analytical and bioanalytical chemistry Vol. 405; no. 20; pp. 6583 - 6587
• Main Authors: Šatínský, Dalibor, Havlíková, Lucie, Solich, Petr
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.08.2013; Springer; Springer Nature B.V
• Subjects: Acetic acid; Acetonitrile; Airborne sensing; Analytical Chemistry; Biochemistry; Biomimetics; Calibration; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatography; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; Crystals; Flow rate; Fluorescence; Food Science; High performance liquid chromatography; Humans; Identification and classification; Laboratory Medicine; Liquid chromatography; Mathematical analysis; Methods; Monitoring/Environmental Analysis; On-line systems; Particle size; Pretreatment; Propranolol - chemistry; Propranolol - urine; Propranolol hydrochloride; Sample preparation; Sensitivity and Specificity; Testing; Time Factors; Urine; Wavelengths; Large-volume sample; Urine; Column switching; Fused-core column; Propranolol; HPLC
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-013-7098-4

A new and fast high-performance liquid chromatography (HPLC) column-switching method using fused-core columns in both dimensions for sample preconcentration and determination of propranolol in human urine has been developed. On-line sample pretreatment and propranolol preconcentration were performed on an Ascentis Express RP-C-18 guard column (5 × 4.6 mm), particle size, 2.7 μm, with mobile phase acetonitrile/water (5:95, v / v ) at a flow rate of 2.0 mL min −1 and at a temperature of 50 °C. Valve switch from pretreatment column to analytical column was set at 4.0 min in a back-flush mode. Separation of propranolol from other endogenous urine compounds was achieved on the fused-core column Ascentis Express RP-Amide (100 × 4.6 mm), particle size, 2.7 μm, with mobile phase acetonitrile/water solution of 0.5 % triethylamine, pH adjusted to 4.5 by means of glacial acetic acid (25:75, v / v ), at a flow rate of 1.0 mL min −1 and at a temperature of 50 °C. Fluorescence excitation/emission detection wavelengths were set at 229/338 nm. A volume of 1,500 μL of filtered urine sample solution was injected directly into the column-switching HPLC system. The total analysis time including on-line sample pretreatment was less than 8 min. The experimentally determined limit of detection of the method was found to be 0.015 ng mL −1 . Figure Chromatogram 1 , which was recorded by direct injection of 1,500 μL of two different urine samples without SPE sample pretreatment. Chromatogram 2 , which was recorded by injection of 1,500 μL of urine sample with propranolol directly to the column-switching system