Design of Experiment Utilization to Develop a Simple and Robust RPUPLC Technique for Stability Indicating Method of Ciprofloxacin Hydrochloride and Metronidazole in Tablets

Abstract

Ciprofloxacin Hydrochloride and Metronidazole are indicated for the treatment of intra-abdominal and
pelvic infections caused by E. coli and diverticulitis. It is necessary to establish a Validated HPLC
method for the assay of them. Using of DOE techniques provides more information about different
factors affects system suitability of new method. DOE led to highly robust methods through creating
design space. The method was performed on Phenomenex Kinetex C18 (50x4.6 mm, 2.6 µm) and the
mobile phase consisted of triethylamine (TEA), (0.5 % v/v, pH 4.5) and acetonitrile which pumped at
a flow rate 0.6 mLmin-1 at 50 °C in gradient manner. 2 µL of drugs sample solutions were monitored
at fixed wavelengths 320 nm. Ciprofloxacin Hydrochloride, Metronidazole and Caduet tablets bulk
powders were stressed under different conditions in forced degradation studies. Major degradation
products were identified such as 2-methyl-4-nitroimidazole, Fluoroquinolonic acid and Ciprofloxacin
Ethylenediamine. The proposed method was validated in terms of linearity, accuracy, precision and
limits of detection and quantitation according to ICH.