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ons had been MedChemExpress INCB3344 plotted by using the median-effect equation: fa ~1=1zDm =Dm where D may be the dose. Dm is the dose required for 50% inhibition effect, which can be equivalent to median-effect dose. fa is definitely the fraction impacted by dose D, and m is really a coefficient in the sigmoidicity in the dose-effect curve. The medium-effect plot is according to the logarithmic kind of Chow's median-effect equation: logfa =fu ~m logD{m logDm where fu is the fraction unaffected, fu ~1{fa. Combination index based on the classic isobologram equation is used for data analysis of two-way combination: Results and Discussion DPPH-scavenging Capacity of the Combined Fractions from AME and PL The DPPH radical-scavenging activity of different combined fractions from AME and PL are shown in Fig.1. Sixteen combined fractions exhibited a wide range of differences in scavenging DPPH free radicals. Among them, the ethyl acetate fraction of Paeonia lactiflora presented the highest potency in scavenging DPPH radical when used in combination with four solvent-extracted fractions from AME, suggesting that EA-PL was rich in DPPH radical-scavenging activity. Our results were in agreement with the previous report. Herein, CIs were used to determine the possible interactive actions between the extracts or fractions. In order to calculate the CIs, dose-effect curves for the single extract or fraction were analyzed. As shown in Fig.1B, the CIs for EA-PL+CFAME and EA-PL+NB-AME were,1.0, indicating these combined extracts had a synergistic effect in scavenging DPPH radical. By contrast, the CIs for several combined fractions were.1.0, suggesting these combinations had antagonistic effects in scavenging DPPH radicals. As is well known, the phenolic and flavonoid In Vitro Synergistic Antioxidant Activity compounds are the most commonly studied substances that greatly contributed to antioxidant activity of plant foods. Therefore, we measured the total phenolic and flavonoid contents in EA-PL+CFAME combination, which exhibited the strongest activity in scavenging DPPH radicals. Results showed that the total phenolic and flavonoid contents were 603.339622.894 mg GAE/g and 121.78561.264 mg RE/g, respectively, which were significantly higher than those of other combinations. These results suggested that the EA-PL fraction had the highest potency in scavenging DPPH radicals to warrant further fractionation. Thus, EA-PL was further chromatographed on a silica gel column using a stepwise elution of methanol/chloroform to afford 8 fractions. Then, each fraction was combined with CF-AME to yield eight AME-PL combined extracts, i.e., E1, E2, E3, E4, E5, E6, E7 and E8. These samples were examined for in vitro antioxidant activity using DPPH free radical scavenging assay and FRAP test. In Vitro Antioxidant Activity of the Eight AME-PL Combined Extracts The antioxidant activity might be attributed to different mechanisms, such as free radical scavenging, reducing potency, prevention of chain initiation, decomposition of peroxides and binding of transition metal ion catalysis. Meanwhile, considering the complexity of the composition of herbal extracts, combined assays are needed for determination of their antioxidant activity. Herein, both DPPH scavenging test and FRAP were applied for the evaluation of antioxidant activity of the AME-PL combined fractions. DPPH Free Radical Scavenging Activity.