An Unbiased Look At CT99021

Total polyphenol content The total polyphenol content of each extract was measured by the method selleck chemical of Folin-Denis (16). Briefly, 0.5 mL of 1 N Folin-Ciocalteu reagent was added to 0.5 mL of each sample. The mixture was allowed to incubate for 3 min and then 10 mL of 2% Na2CO3 solution was added. The resulting solution was mixed, incubated for 1 h, and then the absorbance was measured at 750 nm using a UV/VIS spectrometer (V-570, Jasco Inc., Hachioji, Japan). The total polyphenol content was determined from a tannic acid standard curve. Tyrosinase inhibitory effect The tyrosinase inhibitory effect was measured by the method of Wong et al. (17). A crude tyrosinase solution was prepared by dissolving mushroom tyrosinase in 50 mM sodium phosphate buffer (pH 7.0). Subsequently, 0.2 mL of the crude tyrosinase solution and 0.1 mL of extract were added to 2.8 mL of 10 mM catechol solution. For the control sample, 50 mL of sodium phosphate buffer (pH 7.0) was added to the mixture instead the crude tyrosinase solution. The absorbance of each reaction mixture was measured at 420 nm by a UV/VIS spectrometer (V-570, Jasco Inc.). Ascorbic acid was used as the positive control. Nitrites cavenging effect The method described by Gray and Dugan (18) was used to measure the nitrite PI3K inhibitor scavenging effect of each extract. Briefly, 0.1 mL of 1 mM NaNO2 solution was added to 0.2 mL of each extract, and the pH values of the resulting mixtures were adjusted to 1.2 (0.1 N HCl), 3.0, 4.2, or 6.0 using a 0.2 N of buffer solution. The final volume of each sample was adjusted to 1 mL. The samples were allowed to react at 37��C for 1 h, mixed thoroughly with 5 mL of 2% acetic acid PDGFRB and 0.4 mL of Griess reagent, and then incubated at room temperature for 15 min. The nitrite content of each mixture was determined by measuring the absorbance at 520 nm (SpectraMax M2, Molecular Devices, LLC.). The control was processed with the same method, but was prepared with 0.4 mL of distilled water instead of 0.4 mL Griess reagent. Ascorbic acid was used as the positive control. ACE inhibitory activity ACE inhibitory activity was measured by the modified method of Cushman (19). ACE crude enzyme extract was prepared by adding rabbit lung acetone powder (Sigma, St. Louis, MO, USA) to 300 mM NaCl-100 mM sodium borate buffer (pH 8.3) at a ratio of 0.2 g/10 mL (w/v). The mixture was incubated at 4��C for 24 h and then centrifuged (4��C, 8,000 rpm, 70 min) to obtain ACE crude enzyme extract. To measure ACE inhibitory activity, 100 ��L of 450 mM NaCl-100 mM sodium borate buffer (pH 8.3) and 50 ��L of 5 mM hippuryl-histidyl-leucine were added to 50 ��L of each AG extract, and the mixtures were pre-cultivated at 4��C for 10 min. Next, 50 ��L of the ACE crude enzyme extract was added to each mixture and the mixtures were incubated at 37��C for 30 min.