Accurately weighed ��-CD was dissolved in distilled water to get a saturated solution. Then, Imatinib Mesylate Bcr-Abl the icariin solution in absolute ethanol was added drop by drop and a suspension Inhibitors,Modulators,Libraries was formed. The suspension was agitated for 4 h at 60��C and the ethanol was removed by rotary evaporation. Subsequently, the pH of the reaction system was adjusted and cellulase was added. Finally, the enzymatic hydrolysis experiment could be carried out. Mono-factor experiment for inclusion complex and free icariin The pH value, temperature, ratio of enzyme/substrate in the reaction mixture, concentration of the substrate, and reaction time were fixed at 4.5, 50��C, 1:1, 10 mg/ml, and 24 h, respectively. To evaluate the effect of each factor on enzymatic hydrolysis, the other four factors were fixed and all the reactions were tested in triplicate.

The pH value was investigated, including the values 4.0, 4.5, 5.0, 5.5, 6.0, and 6.5. The temperature Inhibitors,Modulators,Libraries was studied, including 25��C, 37��C, 50��C, 60��C, and 70��C. The ratio of enzyme/substrate in reaction Inhibitors,Modulators,Libraries mixture was inspected, including 1:10, 1:5, 2:5, 3:5, 4:5, and 1:1. The substrate concentration was investigated, including Inhibitors,Modulators,Libraries 1, 5, 10, 20 and 30 mg/ml. Finally, the reaction time was studied, including 2, 4, 6, Inhibitors,Modulators,Libraries 12, 24, and 48 h. The final reaction product was dried by the N 2 blow and dissolved in a specific volume of ethanol. Subsequently, 10 ��l filtered solution was injected into the HPLC system. The content of baohuoside I was analyzed using the single-point external standard method. The transforming rate of icariin in each reaction was used to reflect the enzymatic hydrolysis efficiency.

Preparation of baohuoside Carfilzomib I under optimum condition The inclusion complex and free icariin were transformed into baohuoside I according to the optimized screening conditions. Consequently, the maximum product of baohuoside I was obtained individually for further purification. The produced baohuoside I was extracted from the reaction mixture with ethyl acetate. Subsequently, baohuoside I was dissolved in methanol and subjected to silica gel column using CHCl3�CMeOH for elution. The eluent that only contained baohuoside I was merged. Finally, baohuoside I was obtained through reduced pressure recovery. The identification of produced baohuoside I was carried out by 1H NMR and 13C NMR. RESULTS AND DISCUSSION Differential scanning calorimetry DSC analysis was generally performed to characterize inclusion compounds by comparing the thermal behaviors of the individual components as well as their physical mixtures and inclusion compounds.[21�C23] The DSC curves of icariin, ��-CD, icariin/��-CD physical mixture, and inclusion complex are shown in Figure 3.