본 연구에서는 기존 율피 중에 존재하는 ellagic acid 정 량 할 수 있는 분석법을 확립하였다. 본 실험에서 사용된 밤과 율피, 율피 가루는 2023년 10월 서울 소재 전통 및 약령시장에서 구입하였다. 실험에서 사용된 밤 품종은 국 내에서 소비되는 품종으로 서산, 옥광, 이평, 석추, 중국산 을 구입하여 사용하였고, 가공된 율피와 율피가루의 원산 지는 공주, 아산지역의 제품을 구입하여 실험을 진행했다. 율피중에 존재하는 ellagic acid의 함량은 100% methanol 에서 0.077 mg/g으로 가장 높게 검출되었고, 50% methanol 과 ethanol에서 0.048 mg/g, 증류수에서는 0.011 mg/g로 확 인되었다. 유기용매에 비율이 낮을수록 ellagic acid의 함 량은 순차적으로 줄어듬을 확인하였다. Waters사의 HPLC 이용하였으며 분석용 컬럼은 Aegispak C18-L을 사용하였 고, 이동상은 A (1.2% phosphoric acid in DW)와 B (methanol)를 이용하여 gradient 를 사용하였다. 유속은 1.0 mL/min, 주입량은 10 μL, 컬럼 오븐은 35oC이였다. 검 량선 작성을 위해 1-50 g/mL 범위에서 상관계수(R2) 0.9999 이상의 직선성을 확인하였다. 검량선내 3개 농도를 이용 하여 정밀도와 정확성을 측정한 결과, 일내 정밀도는 0.09- 0.18%, 정확도는 99.9-105.8%, 일간 정밀도는 0.04-0.23%, 정확도는 99.8-105.9% 범위내로 확인되었다. 국내에 유통 되고 있는 시료 8가지 품종의 정보는 Table 10과 같이 정 리하였고, 확보된 시료를 분석한 결과 총 8개 시료에서 ellagic acid의 함량은 0.0002-0.0936 mg/g로 확인되었다. Ellagic acid의 검출 한계는 3개 시료에서 검출한계 0.016 μg/ mL, 정량한계 0.499 μg/mL로 확인되었다. Ellagic acid 측정 불확도 산출 결과 0.40±0.01 mg/kg (신뢰수준 95%, K=2)로 비교적 낮은 측정불확도 값을 산출하였다. 따라서 본 연 구에서는 율피 중 ellagic acid 정성 및 정량분석을 위해 유효성이 검증된 분석법 확립으로 인하여 율피 중 ellagic acid의 기준규격 설정 및 관리에 참고 자료가 될 수 있고, 향후 ellagic acid를 이용한 건강기능식품 개발에 있어 품종, 지역 및 추출용매에 따른 실험결과를 바탕으로 건강기능 식품 기준규격 관리와 활성 및 독성시험 연구의 근거 자 료가 될 수 있다고 판단된다.

In this study, the anti-oxidant, whitening, and anti-wrinkle effects of Castanea crenata inner shell extracts processed by enzyme treatment and pressurized extraction were investigated. The Castanea crenata inner shell was first hydrolyzed using celluclast, viscozyme, or hemicellulase. Then, it was subjected to pressure extraction for different durations (30, 60, and 120 min). The yields of the Castanea crenata inner shell extracts processed by different enzyme treatments followed by pressurized extraction for different times are in the range of 12.42-29.80%. The total polyphenol, flavonoid, and tannin contents of the C30m (celluclast enzyme and autoclave extracts at 30 min) extract were 15.48, 10.82, and 15.82 g/100 g, respectively. The total sugar content of the H120m(hemicellulase enzyme and autoclave extracts at 120 min) extract is 61.07 g/100 g. The 1,1-diphenyl-2-pycrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities of the C30m extract at 1,000 μg/mL are 89.20, and 81.96%, respectively. The superoxide radical scavenging and ferric-reducing antioxidant power of the C30m extract at 1,000 μg/mL are 67.63% and 1,324.79 μM, respectively. Further, the tyrosinase and elastase inhibition activity of the C30m extract at 1,000 μg/mL are 61.32, and 61.06%, respectively. Our results indicate that the Castanea crenata inner shell extracts processed by enzyme treatment followed by pressurized extraction could have beneficial effects on facial skin and they should be considered for use in new functional cosmetics.

Background: There is an increasing surplus of chestnut that are abandoned due to their failure to meet customer awareness. Thus, we investigated the anti-proliferative and apoptotic effects of chestnut (Castanea crenata) inner shell extracts in hepatocarcinoma HepG2 cells as a potential source of anti-cancer materials. Methods and Results: Distilled water extract (CI-W) and ethanol extract (CI-E) were prepared from chestnut inner shell and evaluated their anti-proliferative effects in vitro. Each extract significantly decreased the cell viability of HepG2 cells in a dose- and time-dependent manner. Indeed, the morphology of HepG2 cells treated with CI-W or CI-D was distorted to shrunken cell masses. Furthermore, it was revealed that their extracts induced cell death as evidenced by increased reactive oxygen species (ROS), formation of apoptotic body and condensation. In addition, Their extracts clearly modulated the down regulated of Bcl-2 (anti-apoptoic)/ Bax (pro-apoptotic) family and cleaved caspase-3 as an effector caspase in a dose-dependent manner. Conclusion: These results indicate that the extracts of chestnut inner shell can be used as an anti-proliferative therapeutic agent or functional food.

Atopic dermatitis (AD) is a common chronic inflammatory disease associated with a cutaneous hypersensitivity reaction to an allergen. Although the incidence of AD is increasing these days, therapeutics has yet to be developed for its treatment. The aim of this study was conducted in order to compare and investigate the characteristic between the Castanea crenata inner shell extract (CS) and the Castanea crenata inner shell extract fermented by Lactobacillus bifermentans (FCS) for an anti-atopic medication. The total polyphenol and flavonoid contents were similar to CS and FCS. In the DPPH and superoxide anion radical scavenging, the CS and FCS had the potential for antioxidant activities. Both of them did not exhibit cytotoxicity to HS68 cells. The evaluation of the anti-inflammatory activity in Raw264.7 cells demonstrated that the FCS has inhibited the LPS-induced production of nitric oxide as compared to the CS. The anti-atopic dermatitis test was done through the induction of DNCB in AD hairless mice. The FCS has inhibited the development of the atopic dermatitis-like skin lesion by transdermal water loss, melanin and erythema of the skin as compared to the CS. Moreover, the pro-inflammatory cytokine IL-1β and TNF-α production in hairless mice were inhibited by the FCS treatment. It indicates that the fermentation of the Castanea crenata inner shell has the potential for the treatment of atopic dermatitis.