Influence of Whisky Congeners on Health - The Institute of Brewing ...

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Suppressive effect to melanogenesis Inhibition <strong>of</strong> tyrosinase activity A process <strong>of</strong> aging in the oak cask allows whisky and brandy to contain substances exuded from oak during aging in the cask as well as products made from their interactive or degenerating reactions such as lignin-derived aromatic components and polyphenols. Researchers have reported that some plant elements with a phenol structure inhibit melanin synthesis. The inhibitory effect <strong>of</strong> respective test substances was examined on the enzyme activity <strong>of</strong> tyrosinase prepared from mouse B16 melanoma cells (9) . <strong>Whisky</strong> and brandy had a more potent tyrosinase inhibiting activity than other alcohols (Sake, Shochu, wine, beer) (Fig 11) This inhibitory activity was comparable to one observed in extremely potent plant extracts and increased in proportion to the length <strong>of</strong> storage in the cask. Non-volatile fractions were more active than volatile fractions. (data not shown) brown spirits <strong>Whisky</strong> A <strong>Whisky</strong> B <strong>Whisky</strong> C Brandy A Brandy B Brandy C Shochu A Shochu B Shochu C low Inhibiting activity High kojic acid ( positive control) 0.00 0.01 0.02 0.03 0.04 0.05 1 / IC50 (ml / μg) Figure 11 Inhibitory effects <strong>of</strong> distilled liquors to tyrosinase activity Tyrosinase source was prepared from murin B16 melanoma cells (Riken Cell Bank, Tsukuba, Japan). The cells were lysed by incubating at 4°C for 1hr in RIPA buffer (10 mM Tris-HCl, pH 7.5, 1 % NP-40, 0.1 % sodium deoxycholate,
0.1 % SDS, 150 mM NaCl, 1 mM EDTA, 0.5 mM AEBSF, and 10 μg/mL leupeptin). The lysate were centrifuged at 10,000g for 30 min to obtain the supernatant as the crude tyrosinase extract for the activity assay. The reaction mixture contained 50 mM phosphate buffer, pH 6.8, 0.05 % L-dopa and the supernatant (tyrosinase source). After incubation at 37°C for 20 min, dopachrome formation was monitored by measuring absorbance at wavelength 492 nm in a microplate reader. Effect <strong>of</strong> anti-melanogenesis Inhibitory effect <strong>of</strong> whiskey congener on melanin synthesis was determined in B16 melanoma cell line (Fig 12). We isolated and identified several constituents from whisky congeners and assessed the ability to inhibit melanin synthesis with an ultraviolet-induced pigmentation test system in the skin <strong>of</strong> the colored guinea pig. As a result, lyoniresinol, a polyphenol, was found to be effective in reducing pigmentation. Its activity was approximately 7 times as potent as that <strong>of</strong> arbutin, which was used as a positive control. The test result demonstrated that lyoniresinol did not directly act against the activity <strong>of</strong> tyrosinase (Fig 13). whisky (-) whisky (+) Figure 12 <strong>Whisky</strong> effect <strong>of</strong> anti-melanogenesis in B16 melanoma cell line. Mouse B16 melanoma cells were purchased from Riken Cell Bank (Tsukuba, Japan). B16 melanoma cells were cultured in Dulbecco’s modified eagle’s medium (DMEM) that was supplemented with 10 % (vol/vol) fetal bovine serum, 100 unit/mL penicillin, and 100 μg/mL streptomycin at 37°C in a humidified CO2-controlled (5 %) incubator. Induction <strong>of</strong> melanogenesis was initiated by addition <strong>of</strong> 1 μM [Nle 4 , D-Phe 7 ] α-MSH. 0-200 μg/mL <strong>of</strong> whiskey congener was applied.
Page 1 and 2: Influence
Page 3 and 4: Polyphenols exude from the oak cask
Page 5 and 6: Experimental Purine and carbohydrat
Page 7 and 8: time, and the samples were subjecte
Page 9 and 10: Figure 4 Influence
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Page 13 and 14: ppm. This level and its specific ac
Page 15: sorbitol in erythrocytes (nmol/mLRB
Page 19 and 20: Whisky and brandy
Page 21 and 22: Number of literatu
Page 23 and 24: serum uric acid level (%) ( %) 115
Page 25 and 26: a serum uric acid level (%) b 115 1
Page 27 and 28: purine body xanthine oxydase uric a
Page 29 and 30: whisky Evaporation and lyophilizati
Page 31 and 32: low low inhibiting inhibiting activ
Page 33 and 34: own spirits Whisky
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