Abstract
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•A sensitive and selective method for TDM of Helicobacter pylori triple therapy.•The first method for simultaneous quantitation of OME, TNZ and CLA in human plasma.•An efficient liquid–liquid extraction sample clean-up procedures.•Application for TDM after a single dose of the triple therapy.
The infection of gastric mucosa by Helicobacter pylori (HP) is an essential cofactor in the aetiology of gastroduodenal ulcer and gastric carcinoma. Because of the bacterial resistance, combination therapy containing omeprazole (OME), tinidazole (TNZ) and clarithromycin (CLA) is commonly used for eradication of HP. However, the simultaneous determination of the triple therapy in human plasma was not reported. A simple, reproducible, and selective HPLC method was developed for the simultaneous determination of the triple therapy mixture used for management of HP infections in human plasma. An HPLC procedure based on a liquid–liquid extraction, enrichment of the analytes and subsequent reversed-phase chromatography with UV detection was used. To enable sensitive and selective detection, the method involved the use of online wavelength switching detection, with two different detection wavelengths; 280nm for detection of OME and TNZ and 210nm for detection of CLA. Separations were performed on C18 analytical column with acetonitrile–10mM phosphate buffer of pH=3.0 at flow rate of 1.0mLmin−1. The linear ranges in human plasma were 0.05–10μgmL−1 with correlation coefficients >0.9990. The detection limits in human plasma were 0.02–0.07μgmL−1. Validation parameters were assessed in compliance with US-FDA guidelines. The method proved to be valuable for the therapeutic drug monitoring after oral administration of triple therapy tablets.