An ecofriendly green liquid chromatographic method for simultaneous determination of Amoxicillin, Metronidazole and Ciprofloxacin; application to dosage form and human urine

Green liquid chromatographic method using cyano column and ethanol and 0.5% acetic acid in water (pH 2.8) as mobile phase in gradient mode has been developed for analysis of Amoxicillin, Metronidazole and Ciprofloxacin in human urine. Quantification was carried out using a diode-array UV detector. The detection wavelength was 255 nm. The retention times and detection limits for each antibiotic were 4.9 min and 0.5 μg/mL for Amoxicillin, 6.035 min and 0.14 μg/mL for Metronidazole, 8.03 min and 3.07 μg/mL for Ciprofloxacin, respectively. The developed method was applied to examine the urinary excretion pattern of Amoxicillin, Metronidazole and Ciprofloxacin in healthy male volunteer after an oral administration of Metronidazole and Ciprofloxacin in their combination tablets (Ciprodiazole) and combination tablets of Ciprofloxacin and Amoxicillin (Helicocin). The developed HPLC method was successfully used for the analysis of the selected drugs in their dosage forms and human urine without interference from the excipients or urine matrix. The proposed method was rapid, specific, precise, accurate, environmentally friendly and suitable for bioequivalence and pharmacokinetic studies.