Phellinus Spp Analysis

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Preparation of Phellinus spp extract
All mushroom Phellinus spp used for preparing extract were obtained from company ‘bukseorak’ (Seoul, Korea). Fresh mushrooms were washed and ground with grinder to perform successive extractions. One kilogram of fresh mushroom was incubated in 5 L of distilled water, and filtered under high temperature/ high pressure three times. The filtrate was heated and dried to over 60% concentration using a rotary vacuum evaporator.

Male SD rats (180~200 g, 6 weeks old) were used for the study. After 1 week of adaptation, all animals were housed in a temperature (25℃) and humidity (50%) controlled room with a 12-h light/12-h dark cycle. Water and a normal standard pellet diet were available ad libitum throughout the experimental period.
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The two groups were Group I (control group): rats received 0.2 mL water by gavage, and Group II (Phellinus spp-treated group): the rats were treated with Phellinus spp extract (0.04mg/kg). After or before 30 min, each rat received alcohol diluted in water (40%, v/v) at 5 g/kg. At 1, 3 and 5 h after the administration of alcohol, blood was collected from the tail vein to determine biochemical parameters. The rats received humane care, and experiments were performed according to the criteria outlined in the Guide for the Care and Use of Laboratory Animals and with approval of the Animal Care and Use Committee of Daejeon University (DJUARB2014-046).

Serum alcohol concentration
A blood alcohol assay kit (Abcam, USA) was used, following a slightly modified version of the manufacturer's protocol, to determine the serum alcohol concentration. Briefly, 5 μl of serum was mixed with 50 μl of reaction mixture. After mixing for 30 min at 37℃ the absorbance was measured at a wavelength of 570 nm. The alcohol concentration was calculated according to the equation provided with the kit.

Assessment of AST and