Preparation and characterization of the inclusion complex Preparation of inclusion complex The inclusion complex of icariin with the ��-CD at 1:1 molar ratio was prepared using saturated solution method. Accurately weighed ��-CD was dissolved in distilled water to get a saturated solution. Then, the icariin solution in absolute ethanol was added drop by drop. The formed suspension was selleck Bortezomib agitated for 4 h at 60��C. Subsequently, it was placed at 4��C for 24 h to precipitate the complex. The suspension was filtered and the free icariin was washed from the residue using ethanol. Finally, the precipitation was dried at 100��C for 1 h on the water bath. Preparation of the physical mixture of icariin and ��-CD A physical mixture of icariin and ��-CD in the same weight ratio as the complex was prepared.
Physical mixture was previously sieved into individual components through a 315-��m mesh in a mortar and pestle for 5 min. Differential scanning calorimetry To obtain the differential Inhibitors,Modulators,Libraries scanning calorimetry (DSC) curves of icariin, ��-CD, equimolecular physical mixture of icariin, and inclusion complex, a Photo DSC204 F1 differential scanning calorimeter calibrated Inhibitors,Modulators,Libraries with indium was utilized. Accurately weighed samples (1-5 mg) were placed in a 40-��l aluminum pan, which was sealed and pierced. Alumina was used as a reference material and the scanning rate was 10.0��C/min with a scanning temperature range of 0-450��C. Measurements were made in duplicate for each sample. Solubility studies of inclusion complex Quantification of icariin in the inclusion complex Icariin (98%, 26.
10 mg) was accurately weighted and dissolved in 50 ml purified water to get store solutions. Solutions with volumes of 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 ml were taken out and ethanol was added to each to obtain a volume of 25 ml. The samples were analyzed by the HPLC system using an isocratic elution of CH3OH:H2O (75:25, v/v) as the mobile phase at a flow rate 1.0 ml/min, Inhibitors,Modulators,Libraries and detected at 270 nm wavelength. The regression equation was derived Inhibitors,Modulators,Libraries from the values between the peak area (Y) and the concentrations (X) of icariin. The content of icariin was calculated with the regression equations. Inhibitors,Modulators,Libraries Sample processing Solubility measurements were carried out as follows: Excess amounts of the inclusion complex or free icariin were added to 10 ml of aqueous solutions.
The suspensions were shaken for 72 h at 37��C at Carfilzomib a constant speed of 150 r/min and protected from light to achieve equilibration. After equilibration, the suspensions were filtered through 0.45-��m membranes. The filtrate thus obtained was collected. All procedures were conducted at the specified test temperature to avoid any precipitation of the drug. The filtrate was appropriately diluted with ethanol and the concentration of the substrate (the term ��substrate�� in the whole article refers to icariin) was determined by the HPLC system.