DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR ESTIMATION OF AMOXICILLIN, OMEPRAZOLE AND RIFABUTIN IN SYNTHETIC MIXTURE

Abstract

Helicobacter pylori (H. pylori) infection is one of the most prevalent infectious diseases worldwide, which exists in almost
50% of the world’s population. H. pylori infection plays an important role in gastric adenocarcinoma and the
development of chronic gastritis, gastric ulcer, duodenal ulcer and gastric mucosa-associated lymphoid tissue lymphoma.
A simple, sensitive and rapid isocratic RP-HPLC method has been developed for simultaneous estimation of amoxicillin
(AMX), omeprazole (OMP) and rifabutin (RFB) in synthetic mixture. The analysis was performed on a Thermo C18
analytical column (250 mm × 4.6mm i.d., 5.0μm) with a mobile phase consisting of 20mM KH2PO4: acetonitrile (pH
4.0 with OPA) in the ratio of 20:80v/v. The UV detector was operated at 230mm and the effluents were pumped with a
flow rate of 1.0 ml/min. The run time under the optimum chromatographic a condition is less than 7 min. Linearity,
accuracy and precision were found to be acceptable over the concentration range of 5-25g/ml for AMX and 1-5g/ml
for OMP and RFB. The sensitivity of the method allows the determination of the studied drugs with a limit of
quantification of 1.05, 0.020 and 0.015g/ml for AMX, OMP and RFB respectively. The proposed method was fully
validated according to ICH guidelines. The high sensitivity and the simplicity of the proposed method allow the
successful determination of such a ternary mixture with a percentage recovery of 99.09%±0.260, for AMX,
99.49%±0.163 for OMP and 98.10% ± 0.921 for RFB. The results showed that AMX, OMP and RFB together could be
separated and determined simultaneously with low LOD and LOQ values using the proposed HPLC method. The
method proved to be valuable for the therapeutic drug monitoring after oral administration of ternary mixture.