disease-modifying antirheumatic drug (DMARD) used in the treatment of patients with rheumatoid arthritis. Clinical studies during the last two decades have provided evidence of the drug's efficacy in the treatment of both ulcerative colitis and Crohn's disease [1].Acid dissociation constant values (pK a ) are useful physicochemical measurements describing the extent of ionization of functional groups with respect to pH. This parameter is important in pharmaceutical drug discovery and development [2]. Various experimental modalities have been used for the determination of the pK a values, comprising potentiometric titration [3], spectrophotometry [4], high performance liquid chromatography (HPLC) [5, 6] and capillary electrophoresis (CE) methodology [7]. Among these techniques, most of the drugs can be analyzed by reversed-phase liquid chromatography (RPLC) technique because of the several advantages like rapidity, specificity, accuracy, precision and ease of automation in this method.Analysis of the change in retention time of the analyte versus the change in pH of the mobile phase is given an indirect measure of the acid dissociation constant. The theory for studying the pH dependence of chromatographic retention for ionizable compounds in LC was proposed by Horvath et al. and verified by different authors for different type of compounds [8-10]. For a triprotic acid (H 3 A), Eqs. (1), (2) and (3) represent the variation of the retention factors at different pH values.(Abstract Acid dissociation constant value (pK a ) is key parameter for predicting the extent of the ionization of a drug molecule at different pH. The effect of the mobile phase composition on the acid dissociation constant values was studied by measuring the pK a values at different acetonitrile concentrations, ranging from 25 to 35 % (v/v). pK a values and limiting retention factors of SSZ were calculated chromatographically using equations derived. From calculated pK a values, the aqueous pK a values of SSZ were calculated by various approaches. Moreover, the correlation established between retention factors and the pH of the water-acetonitrile mobile phase was used to determine the optimum separation condition. Optimized condition was also validated with respect to system suitability, linearity, precision, accuracy, limit of detection and limit of quantitation, respectively. A Gemini NX C18 column (250 × 4.6 mm I.D., 5 µm particles) was used for all the determinations.
Imidazoles are well known heterocyclic compounds having important feature of a variety of medicinal agents. Studies with imidazole containing compounds are great importance in the clinical use of drugs. The presence of imidazole group in the molecules of lanoconazole (LCZ) and butoconazole (BCZ) is very important for its chemical and analytical behavior. Target analytes were efficiently separated with a Pinnacle DB Cyano column (5 µm, 4.6 x 250 mm I.D.) and oxiconazole (OCZ) used as an internal standard. In this study, correlation between the retention factors of the solutes and pH of the mobile phase was determined at ten pH values in acetonitrile-water binary mixture. In optimization condition, quantification of these drugs were performed in human urine. The linear responses were observed over a wide concentration range (6.25.
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