Quantitative determination of propranolol in human serum by high-performance thin-layer chromatography

Abstract

A high-performance thin-layer chromatography (HPTLC) method for the quantification of propranolol in human serum is developed and validated. The method includes a liquid-liquid extraction of the analyte from the matrix using n-heptane-isoamyl alcohol (98.5:1.5, v/v) as the extraction solvent and verapamil as the internal standard.The HPTLC method employed precoated silica gel F254 HPTLC plates (10 cm × 10 cm and 10 cm × 20 cm; layer thickness 0.2 mm) prewashed with methanol, and a mobile phase comprising chloroform-methanol-ammonia (9:1:0.04, v/v/v). The developing solvent was run up to 80 mm in a vertical CAMAG chamber previously saturated with the solvent mixture for 20 min. Densitometric detection was done at λ = 290 nm. The method was linear between 5 and 100 ng/band, corresponding to 0.01 and 0.20 ng μL−1 of propranolol in human serum after the extraction process and applying 50 μL to the chromatographic plates (correlation coefficient = 0.998). The %RSD (absolute value of the coefficient of variatio...