Elix 03

The following were dissolved in a 50 mL beaker: 6.7 mL of acetone, IL (5, 8, or 12 mg), fluconazole (5 mg), and MCT (40 μL). The mixture was subjected to magnetic stirring at 40 °C for 1 h. Separately, 13.3 mL of water was placed in a beaker under magnetic stirring at 25 °C. The fully solubilized organic phase was then slowly poured through a funnel and under moderate stirring (900 rpm) over the aqueous phase, at 25 °C. The emulsions were kept under moderate stirring for 15 min at 25 °C. Thereafter, the organic solvent (acetone) was evaporated from the emulsion under reduced pressure (60 rpm and 40 °C thermostatic bath), reaching a final volume of 10 mL. Three emulsions were prepared containing [C₁₂C₁im]Br or [C₁₆C₁im]Br at concentrations of 5, 8, or 12 mg. We used reduced amounts of surfactant, since the formation of emulsions with these amounts of IL has already been observed. Additionally, at these quantities of IL, it is guaranteed that all of the IL will be solubilized. We chose concentrations below (5 and 8 mg) and above (12 mg) the CMC of the IL [C₁₆C₁im]Br, in order to compare the effect of this property on the system’s behavior. The ultrapure water used to prepare the solutions was double-distilled and deionized using Millipore quality water (Elix-03, Barueri, Brazil; and Milli-Q, Molsheim, France).

HPLC system from WATERS (Milford, USA) is equipped with 515 HPLC pump as a solvent delivery system, rheodyne injection valve with a 20 µL loop and WATERS 2998 photodiode array (PDA) detector set at a wavelength range of 190–400 nm. Separation was performed on an Agilent ZORBAX Eclipse XDB “C₁₈” column (4.6 mm×150 mm, 5 µm). Chromatographic data were recorded and processed using EMPOWER-2 software.
Dissolution system from LABINDIA Disso 2000 is equipped with high precision multichannel pump and sample collector. For weighing analytical balance (Mettler Toledo and Sartorius) and pH measurement, pH meter (Mettler Toledo) was used. HPLC grade water was obtained from water purification systems, Milli-Q, ELIX 03 (MILLIPORE, Milford, MA, USA).
UV-vis double beam spectrophotometer Perkin-Elmer Lambda-35 was used for all spectrophotometric measurements (i.e., for NASSAM), having a slit width of 1 nm, installed with UV-Winlab and UV-Winlab data processor and viewer software. All spectra were saved in comma separated file (CSV) format and then data were statistically analyzed using unscramble 10.2.

HPLC grade acetonitrile (ACN), potassium di-hydrogen phosphate buffer (KH₂PO₄), orthophosphoric acid (OPA) and hydrochloric acid (HCl) were purchased from Rankem (New Delhi, India). HPLC grade water for chromatography and dissolution [obtained from water purification systems Milli-Q, ELIX 03 (MILLIPORE, Milford, MA, USA)] was used. Solutions were filtered through a qualisil nylon syringe filter (25 mm×0.45 µm) Ultipor^® N66^® and membrane filter (47 mm×0.45 µm) (Pall Pvt. Ltd., India) prior to use. Standards of PCM, CPM and PH were procured from Syncom Health Care (Dehradun, India). Marketed formulation named as SNEEZY tablets was labeled as each uncoated tablet contained 500 mg of PCM, 5 mg of PH and 2 mg of CPM (quinoline yellow) batch no. SC12030, manufactured by Cadila Pharmaceuticals, and COLD-GO which is an oral suspension was labeled as each 5 mL of oral suspension contained 125 mg of PCM, 2 mg of CPM and 25 mg of PH (having coloring agent ponceau4R) batch no. HG-278, manufactured by Torque Pharmaceutical Limited, which was procured from local market (Moga, Punjab, India).