ULTRA HPLC METHOD FOR THE SIMULTANEOUS ANALYSIS OF DRUGS AND FLAVONOIDS IN HUMAN URINE

Abstract

A validated reverse-phase Ultra HPLC method for the simultaneous determination of drugs sotalol, metoprolol, propranolol, carvedilol, salicylic acid, dexamethasone, prednisolone, and ketoprofen and flavonoids: (±)-catechin, (−)-epicatechin, rutin, hesperidin, neohesperidin, quercitrin, (±)-naringenin, hesperetin in human urine has been developed. Urine samples were pretreated by solid-phase extraction using SDB and C18 cartridges. The extraction efficiencies of each analyte from urine ranged from 76.21% to 101.29%. Gradient separation is achieved by using a Chromolith® Fast Gradient Monolithic C18e (50 mm × 2 mm) column and Hypersil Gold (50 mm × 2.1 mm, 1.9 µm) column, using UV detection to monitor the analytes at 227, 240, 254, and 280 nm. The mobile phase consists of mixed 0.05% trifluoroacetic acid in water and acetonitrile in the gradient elution. All sixteen compounds were analyzed within 4 min. The LOD and LOQ of drugs and flavonoids are 0.01 µg/mL and 0.03 µg/mL; 0.02 µg/mL and 0.07 µg/mL, respectively. The developed procedure allows the determination of drugs in urine in concentrations from 0.2 to 40 µg/mL. The following concentrations of the examined drugs in human urine, belonging to people treated with β-blockes, were detected. What is more, an amount of flavonoids occur in urine in higher concentrations than calculated LOD and LOQ. The method has been proved to be precise, accurate, and well suited to the routine determination of all these flavonoid and drug concentrations in human urine samples.