[PubMed] [Google Scholar] 2. through the flowering intervals of these plant life. The collected plant life were identified by Dr taxonomically. Khaled Tawaha (Faculty of Pharmacy, Jordan College or university), and voucher specimens had been deposited on the Herbarium Museum from the Faculty of Pharmacy, Jordan College or university of Technology and Research. The plant components had been cleaned out of residual garden soil, air-dried at area temperature, surface to an excellent powder utilizing a lab mill, and passed through a 24-mesh sieve to create a homogeneous natural powder finally. The powder components had been kept in a dark place, at area temperatures (22C23C), until removal. Plant removal Methanolic extractions had been conducted utilizing a 250-mg test of each surface plant material, from the utilized parts [Desk 1], in 10 ml methanol (80%), at 37C for 3 h, within a shaking drinking water bath. After air conditioning, the remove was centrifuged at 1500 for 10 min, as well as the supernatant was retrieved. The solvent was evaporated under vacuum at 40C utilizing a rotary evaporator. The solid residues were stored and collected within a dried out condition until analysis.[20] Table 1 Xanthine oxidase inhibitory activities of the methanolic extracts of 24 medicinal plants collected from different locations in Jordan Open in a separate window XO assay The XO inhibitory activity was measured as previously reported.[16,17,21C23] The substrate and the enzyme solutions were prepared immediately before use. The reaction mixture contained an 80 mM sodium pyrophosphate buffer (pH = 8.5), 0.120 mM xanthine, and 0.1 unit of XO. The absorption at 295 nm, indicating the formation of uric acid at 25C, was monitored and the initial rate was calculated. The methanolic dried extract, initially dissolved and diluted in the buffer, was incorporated in the enzyme assay to assess its inhibitory activity at a final concentration of 200 g/ml. Moreover, for the plants whose extracts showed enzymatic inhibition more than 35%, the IC 50 evaluation was also performed. In this case, five different concentrations of the dried extract (18.8, 37.5, 75, 150, and 300 g/ml) were used to determine the Zileuton concentration that inhibits 50% of the XO enzyme activity (IC50 value). All assays were run in triplicate; thus, inhibition percentages are the mean of three observations. A negative control (blank; 0% XO inhibition activity) was prepared containing the assay mixture without the extract. Allopurinol was used as a positive control in the assay mixture. The XO inhibitory activity was expressed as the percentage inhibition of XO in the above-mentioned assay mixture system, calculated as follows:[21C23] where test inclination is the linear change in the absorbance per minute of the test material, and blank inclination is the linear change in the absorbance per minute of the blank. RESULTS AND DISCUSSION In this study, the methanolic extracts of 23 different plants belonging to 12 different families were investigated as potential XO inhibitors. The selected plants and their XO inhibition assay results are summarized in Table 1. The degree of XO inhibition was evaluated for the extracts of all species at a concentration of 200 g/ml. However, the IC 50 values were determined only for those plants (n = 6) which showed an inhibitory activity of 40% or more when compared to uninhibited enzymatic Zileuton reaction. These latter plants are.Four flavonoids and Zileuton three other constituents from Achillea biebersteinii. g/ml), L. (199.5 g/ml), Afansiev (360.0 g/ml), L. (650.0 g/ml), and L. (595.8 g/ml). Moreover, four more plants, namely Mill. (28.7% inhibition), (L.) Mill. (28.4%), (L.) Kostel. (25.1%), and L. (22.5%) showed a XO inhibitory activity in the range of 22C30%. Conclusion: The study showed that many of the tested plant species are potential sources of natural XO inhibitors that can be developed, upon further investigation, into successful herbal drugs for treatment of gout and other XO-related disorders. = 23), were collected from different geographical places of Jordan, during the flowering periods of these plants. The collected plants were identified taxonomically by Dr. Khaled Tawaha (Faculty of Pharmacy, Jordan University), and voucher specimens were deposited at the Herbarium Museum of the Faculty of Pharmacy, Jordan University of Science and Technology. The plant materials were cleaned of residual soil, air-dried at room temperature, ground to a fine powder using a laboratory mill, and finally passed through a 24-mesh sieve to generate a homogeneous powder. The powder materials were stored in a dark place, at room temperature (22C23C), until extraction. Plant extraction Methanolic extractions were conducted using a 250-mg sample of each ground plant material, of the used parts [Table 1], in 10 ml methanol (80%), at 37C for 3 h, in a shaking water bath. After cooling, the extract was centrifuged at 1500 for 10 min, and the supernatant was recovered. The solvent was evaporated under vacuum at 40C using a rotary evaporator. The solid residues were collected and stored in a dry condition until analysis.[20] Table 1 Xanthine oxidase inhibitory activities of the methanolic extracts of 24 medicinal plants collected from different locations in Jordan Open in a separate window XO assay The XO inhibitory activity was measured as previously reported.[16,17,21C23] The substrate and the enzyme solutions were prepared immediately before use. The reaction mixture contained an 80 mM sodium pyrophosphate buffer (pH = 8.5), 0.120 mM xanthine, and 0.1 unit of XO. The absorption at 295 nm, indicating the formation of uric acid at 25C, was monitored and the initial rate was calculated. The methanolic dried extract, initially dissolved and diluted in the buffer, was incorporated in the enzyme assay to assess its inhibitory activity at a final concentration of 200 g/ml. Moreover, for the plants whose extracts showed enzymatic inhibition more than 35%, the IC 50 evaluation was also performed. In this case, five different concentrations of the dried extract (18.8, 37.5, 75, 150, and 300 g/ml) were used to determine the concentration that inhibits 50% of the XO enzyme activity (IC50 value). All assays were run in triplicate; thus, inhibition percentages are the mean of three observations. A negative control (blank; 0% XO inhibition activity) was prepared containing the assay mixture without the extract. Allopurinol was used as a positive control in the assay mixture. The XO inhibitory activity was expressed as the percentage inhibition of XO in the above-mentioned assay mixture system, calculated as follows:[21C23] where test inclination is the linear change in the absorbance per minute of the test material, and blank inclination is the linear change in the absorbance per minute of the blank. RESULTS AND DISCUSSION In this study, the methanolic extracts of 23 different plants belonging to 12 different Mouse monoclonal to SYP families were investigated as potential XO inhibitors. The selected plants and their XO inhibition assay results are summarized in Table 1. The degree of XO inhibition was evaluated for the extracts of all species at a concentration of 200 g/ml. However, the IC 50 values were determined only for those plants (n = 6) which showed an inhibitory activity of 40% or more when compared to uninhibited enzymatic reaction. These latter plants are L. (Lamiaceae), L. (Asteraceae), Reut. ex Boiss. (Asteraceae), Afansiev (Asteraceae), L. (Lamiaceace), and L. (Ginkgoaceae). The inhibitory profiles of three selected examples of these plants are shown in Figure 1. Open in a separate.