Development of high performance liquid chromatography (HPLC) method for simultaneous analysis of metronidazole, miconazole and lidocaine from pharmaceutical preparations

In this thesis, high performance liquid chromatography (HPLC) for simultaneous analysis of the pharmaceutical product containing Metronidazole, Miconazole and Lidocaine active ingredients used in the treatment of candidal vulvovaginitis caused by Candida albicans, bacterial vaginosis caused by Gardnerella vaginalis and anaerobic bacteria, and trichomonal vaginitis caused by Trichomonas vaginalis and mixed vaginal infections, method development and validation studies were carried out with. In the analysis method, ACE C18 250 x 4.6-5µm column was preferred as the stationary phase, and the analytes were eluted with the 35 mM Ammonium Acetate and Methanol gradient system preferred as the mobile phase at a flow rate of 1.3mL/min. İn the study 230 nm wavelength was carried out at with the ultraviolet detector. Metronidazole, Lidocaine, Miconazole, retention times are 3.3 min, 12.1 min and 17.3 min, respectively. The developed methods were based on ICH Q2 (R1) analytical method validation parameters, which are stability, specificity, linearity, accuracy, recovery, limit of detection (LOD), limit of quantification (LOQ), precision, repeatability, robustness, ruggedness parameters. Linearity ranges for all three substances were found as 75 µg/mL – 1125 µg/mL for Metronidazole, 10 µg/mL – 150 µg/mL for Lidocaine HCl, 20 µg/mL – 300 µg/mL for Miconazole nitrate. Limit of detection (LOD) were calculated as 0.39 µg/mL for Metronidazole, 1.72 µg/mL for Lidocaine HCl, 0.82 µg/mL for Miconazole nitrate, while the limit of quantification (LOQ) was detected that 1.17 µg/mL for Metronidazole, 5.21 µg/mL for Lidocaine HCl, and 2.47 µg/mL for Miconazole nitrate. As a result of the validation studies for developed method, it has been found to be accurate, precise, original, sensitive, consistent and robust. With the validated method, the pharmaceutical product containing all three active substances obtained from the market was analyzed and the reliability of the amount of active substance required in a unit dose was proven.