American Journal of Advanced Drug Delivery Open Access

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Abstract

Novel SPE-HPLC Method for Analyses of �?Ž�?²-Blockers in Human Plasma Using New Generation Phenyl-Ethyl Column

Imran Ali, Umma Kulsum, Kishwar Saleem, Norikaju Nagae and Vinay D. Gaitonde

Objectives: The mixture of six β-blockers was analyzed in human plasma by SPE and HPLC methods.

Methods: Timolol, atenolol, oxprenolol, alprenolol, acebutolol and carazolol were separated using Phenyl-Ethyl column (PhE, 250 x 4.6 mm, 5.0 μm). The mobile phase was phosphate buffer (50 mM, pH 3.0)-acetonitrile (70:30, v/v) at 1.0 mL/min flow rate. The detection was set at 220 nm and 27±1ºC temperature.

Results: The values of chromatographic parameters i.e. capacity (k), separation (α) and resolution factor (Rs) were in the range of 2.76- 17.72, 1.14-1.82 and 2.11-6.64, respectively. The limits of detection and quantification were 0.1-0.5 μg/mL and 0.6-0.30 μg/mL, respectively. The percentage recoveries of extraction were in the range of 32.30- 50.5%. π-π, hydrophobic and polar interactions between β-blockers and phenyl-ethyl column were responsible for the separation. The reported SPE and HPLC methods were efficient, reproducible, fast and selective. The developed and validated methods were applied for monitoring six β-blockers in human plasma. Briefly, the developed SPE and HPLC methods can be applied to monitor the reported β-blockers in any sample of biological, environmental and industrial origins.