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72249083018
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2-O).
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72249091790
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2 (3) in the filtrate was concentrated under low pressure, and precipitated with cold diethyl either. The white powder was further washed with diethyl ether and dried in vacuo.
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72249097413
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2 (3) by dropwise. Then, its pH was adjusted to 6.0-7.0 with aq NaOH solution (0.25 N). The metal complexes (4, 5) were completely formed at rt within 3 h. The final products were collected by filtration and washed intensively with distilled water to remove salts.
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0003793129
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Dwyer F.P., and Mellor D.P. (Eds), Academic Press, New York
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24
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72249121559
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Mushroom tyrosinase inhibition assay: Seventy microliters of phosphate buffer (pH 6.8, 0.1 M) containing inhibitor dissolved in DMSO (10 μl) were mixed with 10 μl of 100 μg/ml mushroom tyrosinase, and then 10 μl of 10 mM l-DOPA were added. Following incubation at 37 °C, absorbance was measured every 10 min for at least 1 h at 475 nm. The % inhibitory activity was calculated by the following formula; (A - B)/A × 100 (A: absorbance of reference solution; B: absorbance of test sample solution). Each experiment was performed in triplicate and averaged.
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72249121336
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Crystal violet assay: cell viability was determined using a crystal violet assay (Dooley et al. 1994), After incubation with the samples for 24 h, culture media were removed and replaced with 0.1% crystal violet in 10% ethanol. Mel-Ab cells were then stained for 5 min at room temperature and rinsed four times. The crystal violet retained by adherent cells was then extracted with 95% ethanol, and absorbance was determined at 590 nm using an ELISA reader (TECAN, Salzburg, Austria). Each experiment was performed in triplicate and averaged.
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2. Effects of the inhibitors on melanin formation in Mel-Ab cell line were estimated. When the samples were treated at the concentrations of 10 μM and 100 μM for four days, the inhibitors-treated cells produced much less amount of melanin than the cells without inhibitors. The melanin contents of Mel-Ab cells were measured spectrophotometrically in the experimental period. Each experiment was performed in triplicate and averaged.
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