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Data was represented as the average (mean) of five individual readings and its standard deviation (SD). In vitro stability of labeled complex To ensure the stability of radiolabeled drug complex for entire period of time of experiment, a 100 ��L aliquot of the labeled formulation was mixed with 2.0 mL of phosphate-buffered saline (pH 7.4) and incubated at room temperature for a period of 24 h. Labeling efficiency of the drug �C Tc-99m complex was measured in between to ensure stable complex. Preparation of in situ gel formulation Different combinations of placebo formulations were developed Vemurafenib solubility dmso and screened for viability [Table 1]. Chitosan (viscosity 175 cps, insolubles 0.20%, deacetylation 84%, pH 8.34, moisture 7%, ash 0.48%, total plate count Ruxolitinib in vivo acid and pH adjusted to 5.5�C6.0 by phosphate buffer system. Gellan gum was dissolved in phosphate-buffered saline, pH 7.4. Viscosity was measured using Brookfield's viscometer (model DV II, spindle no. 02, at 20 rpm), and clarity was examined by visual inspection. Different combinations were tested, and the best combination is chosen for further developing medicated in situ gel. The formulations were selected/rejected on the basis of their viscosity, clarity, and turbidity with change in pH. Sparfloxacin is available as 0.5% eye drops concentration for antibacterial activity. Hence, a dose of 0.5% sparfloxacin was used in in situ gel formulation for proper comparison. The weighed quantity of drug was placed in volumetric flask and dissolved primarily in 1�C2 drops of ethanol and then volume was made up in normal saline under aseptic conditions. 0.1% of methyl paraben is approved for ocular use, so same was added as a preservative.[11] Amiloride The resultant solutions were mixed well and kept undisturbed at room temperature for 24 h. Osmolarity of the formulation was determined by osmometer (Fiske Associate, USA) and required amount of sodium chloride was added and mixed thoroughly to make the solution isotonic. Complete formula of the developed formulation is given in Table 2. The other physicochemical evaluation parameters that is, clarity, gelation pH, osmolarity etc., were also measured as per previously available literature.[8,12] The plain drug solution of drug was prepared in the same manner removing polymers (chitosan and gellan gum) and make it isotonic. Table 1 Different combinations evaluated for placebo in situ hydrogel formulation of sparfloxacin Table 2 Ingredients of medicated formulation In vitro drug release profile In vitro release was determined by modified dialysis membrane technique. A 1 ml aliquots of the formulation and marketed formulation were taken in the dialysis tube (Sigma Chemicals, USA), which was suspended in separate beakers. Care was taken to make sure that no air bubbles were entrapped inside the polymer solutions. The beakers were then filled with 100 ml of simulated tear fluid, pH 7.