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2398 J. Med. Plants Res. Table 1. Total phenonic compounds, degree of hydrolysis and FRAP value. RRHs Frap Value (umol/g) DH Total Phenonic Content GAE (ug/g) RRHsI 15.96 ± 0.30 46.69 ± 0.60 63.13 ± 0.84 RRHsII 12.54 ± 0.17 45.31 ± 0.70 60.14 ± 0.23 RRHsIII 20.74 ± 0.80 45.54 ± 0.13 66.01 ± 0.55 RRHsIV 34.01 ± 0.22 49.07 ± 0.15 56.62 ± 0.36 RRHsV 9.87 ± 0.45 38.17 ± 0.02 50.11 ± 0.47 RRHsI, RRHsII, RRHsIII, RRHsIV and RRHsV represent that RRHs are fermented by A. oryzae, R. oligosporrus, M. racemosus, A. niger and P. glaucum, respectively. Each value is expressed as mean ± standard deviation (n = 3). quench the long-lived ABTS radical cation (ABTS· + ), a blue-green chromophore with characteristic absorption at 734 nm. The addition of antioxidants to the preformed radical cation reduces it to ABTS, determining a decolorization. A stable stock solution of ABTS· + was produced by reacting a 7 mmol/L aqueous solution of ABTS with 2.45 mmol/L potassium persulfate (final concentration) and allowing the mixture to stand in the dark at room temperature for 12 to 16 h before use. At the beginning of the analysis day, an ABTS· + working solution was obtained by the dilution in ethanol of the stock solution to an AC of 0.70 ± 0.02 AU at 734 nm (Pellegrini et al., 2003). All the tests were conducted in triplicate. The percentage inhibition of the ABTS radical by the samples was calculated according to the formula: % inhibition= [(A C ) – A A /A C ] × 100 Where A C is the absorbance of the control, A A is the absorbance of the antioxidant sample. Statistical analysis All the data were express as means ± standard deviations (SD) from three independent replicates. Results were evaluated by analysis of variance (ANOVA) with MINTAB 16. Difference was considered significant when P-value was < 0.05. RESULTS AND DISCUSSION Total phenolic compounds It has long been reported that natural phytochemicals in fruits and vegetables have antioxidant activity. Among those substances, the phenolic compounds widely distributed in fruits and vegetables have the ability to scavenge free radicals, superoxide and hydroxyl radicals by single-electron transfer (Li et al., 2005). The total phenonic contents of RRHs are shown in Table 1. Among the five different RRHs, RRHsIII displayed the highest total phenonic content (P < 0.05), followed by RRHs I, RRHsII, RRHIV and RRHsV. Hence RRHsIII had higher reducing power than RRHs I, RRHsII and RRHsV. Interestingly, the total phenolic content of RRHsIV was lower, whereas its reducing power (FRAP value = 34.01 ± 0.22; P < 0.05) was highest for all the RRHs. Therefore, there was no correlation (R 2 = 2.6%) between reducing power and total phenonic content in the five RRHs. Although, some researchers reported that FRAP value were highly correlated with phenol content (Rodríguez et al., 2010; Wang and Lin 2000), our result is in agreement with many other studies. Sun and Ho, 2005) reported that there was no correlation found between the AAC value tested by the bcarotene bleaching method and total phenolics content, nor between the antioxidant activity tested by the Rancimat method and total phenolics content. Free phenolic compounds and antioxidant capacity in some of vegetables exhibited a positive, but not very strong (Chu et al., 2002). The ambiguous relationship between phenolic compounds and antioxidant activity can be explained as follows: (1) phenolic compounds cannot include all the antioxidants (Kahkonen et al., 2001), the antioxidant activity observed could possibly be due to existence of some other antioxidants in RRHs, such as low molecular peptides, oligosaccharides, organic acid and Maillard reaction products, etc. (2) different method to measure antioxidant activity with various mechanisms may lead to different observations. Five different RRHs may contain various complex antioxidant components, which have different antioxidant potency. Antioxidant activities of RRHs Determination by FRAP assay The FRAP assay is simple, precise, sensitive and inexpensive, and gives fast and reproducible results. It is widely used for evaluating both individual antioxidants and their mixtures. The reducing power of five types of RRHs determined at 593 nm is shown in Figure 1. In this assay, the higher optical density (OD) value at 593 nm indicated the higher reducing power. As shown in Figure 1, the concentration-dependent profile of reducing power was obvious for all the tested RRHs. Regarding the five RRHs, RRHsIV showed the highest activity apparently (P < 0.05), followed by RRHsIII, RRHsI, RRHsII and RRHsV. As we know, A. niger is one of important strain of molds in fermentation industry, and is used today in various industrial processes for the manufacture of citric acid (Schreferl-Kunar et al., 1989; Kurbanoglu and Kurbanoglu 2004). It has been reported that citric acid is
DPPH• scavenging ratio(%) OD value at 593nm Figure 1. Reducing power of five typs of RRHs. Each value is the mean ± SD of triplicate measurements. 70 60 50 40 30 20 10 0.7 0.6 0.5 0.4 0.3 0.2 0.1 Figure 2. DPPH redical scavenging of five typs of RRHs. Each value is the mean ± SD of triplicate measurements. a kind of metal chelator, which deactivate trace metals that are free or salts of fatty acids by the formation of complex ions or coordination compounds, thereby terminate radical chain reactions. Lee and Lee (2002) reported that citric acid was used as antioxidant to prevent browning in plant tissue culture. In previous study, our data showed that total acid content of RRHsIV was the highest (0.30%; P < 0.05) followed by RRHsII (0.16%), RRHsIII (0.09%), RRHsI (0.04%) and RRHsV (0.03%). The organic acids (citric acid, gluconic acid, gallic acid etc) in RRHsIV probable were the major contributors to the antioxidant capacity. On the other hand, DH of RRHsIV was 49.07%, which is the highest (P < 0.05) for all the RRHs as shown in Table 1. It was implicated that RRHsIV might contain more low molecular peptides, which had antioxidant activity. A large number of literatures have reported that antioxidant peptides were 0 0 0 0.2 0.4 0.6 0.8 1 Concentration(mg/ml) 0 5 10 15 20 25 Concentration(mg/ml) RRHsI RRHsII RRHsIII RRHsIV RRHsV RRHsI RRHsII RRHsIII RRHsIV RRHsV Tian et al. 2399 purified from protein hydrolysates, and the sequences of amino acid were discovered (Chen et al., 1996; Anisimov et al., 1997; Je et al., 2005). The antioxidant peptides in RRHsIV need further study. Determination by DPPH assay The DPPH method is a valid, easy, accurate, sensitive and economic method to evaluate scavenging activity of antioxidants of fruits and vegetables juices or extracts. When the antioxidants that can provide hydrogen exist, they donate hydrogen to the free radicals so that the radicals remove the odd electron to turn to unreactive ones (Wang et al., 2006). The DPPH radical scavenging effects of five RRHs are shown in Figure 2. The DPPH· scavenging ratio was positively and significantly correlated with concentration. From RRHsI to RRHsV,
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Journal of Medicinal Plants Researc
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Editors Prof. Akah Peter Achunike E
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Electronic submission of manuscript
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Fees and Charges: Authors are requi
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Table of Contents: Volume 6 Number
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Figure 1. Chuanxiong Rhizoma (http:
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Table 1. Identification of main pea
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Table 3. Pharmacokinetic parameters
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active component study, many invest
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and 1185 kg ha -1 in the first site
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Table 1. Recommendation for use fer
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Sadanandan AK, Hamza S (1996c). Stu
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and it has been traced to South Ame
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Table 3. Results of phytochemical a
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(a) (b) (c) Figure 1. (a) Normal ce
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emedies in the south of Brazil. J.
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known fatty acids and terpenoids we
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Table 1. Renal function tests of ra
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Al-Radahe et al. 2271 Figure 3.nnnn
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namely disruption of the vascular e
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Zayachkivska OS, Konturek SJ, Drozd
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β-carotene were purchased from Sig
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Figure 1. Effect of ethanol concent
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Table 3. Climatic and soil factors
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content of polysaccharides. Phospha
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Serge et al. 2285 Table 1. Relative
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Percentage inhibition % of inhibiti
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Table 1. Result of phytochemical sc
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Table 3. Result of thin layer chrom
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Table 2. MIC of a compound 1 from c
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Zirihi et al. 2301 Figure 2. Termin
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Figure 5. T. catappa Linn. (Combret
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Figure 8. A. fumigatus: Aspect of c
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Figure 12. Sensitivity of A. fumiga
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Noormi et al. 2311 Table 1. Callus
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Table 2. Contd. NoC=No callus. 4.0
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Figure 2. Contd. at 2.0 mg/L yellow
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Table 1. Biochemical parameters in
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Figure 3. Bcl-xL reactions in inter
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ACKNOWLEDGMENTS The authors would l
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Figure 1. The first page of the boo
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Figure 3. The most widely included
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Figure 4. Lithographic print “St.
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Table 1. List of plants included in
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Table 1. Contd. Nedelcheva 2333 Cin
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Table 1. Contd. Nedelcheva 2335 Rhe
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14 7 1 15 1 35 17 16 Imported plant
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Academy of Science Publishing House
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et al., 1996; van Wyk and Gericke,
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DPPH inhibition (%) DPPH % inhibiti
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% Mortality Concentration (µg/ml)
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2460 J. Med. Plants Res. Department
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2466 J. Med. Plants Res. AOAC (2000
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2470 J. Med. Plants Res. Table 1. F
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2486 J. Med. Plants Res. components
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2490 J. Med. Plants Res. Inhibition
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2500 J. Med. Plants Res. Flowering
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2524 J. Med. Plants Res. ACKNOWLEDG
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UPCOMING CONFERENCES 15th European
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