호프 추출물의 원료 표준화를 위한 지표성분 Xanthohumol 분석법 유효성 검증

류가희,남민지,최재환,윤숭섭,심지원,신재욱,정운희,이경봉,권동주 한국식품영양과학회 2024 한국식품영양과학회지 Vol.53 No.2

본 연구는 호프 추출물을 건강기능식품 개별인정원료로 개발하기 위해 원료 표준화를 위한 지표성분으로 xanthohumol을 설정하고 이에 대한 HPLC 분석법 유효성 검증을 실시하였다. 분석법의 유효성 검증은 식품의약품안전처 건강기능식품 기능성 원료 인정을 위한 제출자료 작성 가이드와 ICH 가이드라인에 근거하여 특이성, 직선성, 정확성, 정밀성, 검출한계 및 정량한계를 통해 신뢰성 및 타당성을 검증하였다. 지표성분 xanthohumol 분석을 위해 0.1% formic acid가 첨가된 water 및 95% acetonitrile을 이동상으로 사용하였고 컬럼은 Capcell pak C18을 사용하여 HPLC를 이용하여 분석하였다. 표준용액과 호프 추출물의 지표성분 xanthohumol의 피크 머무름시간은 일치하고 다른 물질의 간섭을 받지 않는 것을 확인했으며, UV/visible spectrum이 일치하므로 특이성을 확인하였다. Xanthohumol 검량선의 상관계수(R2)는 0.9999 이상으로 나타나 높은 직선성을 보여 분석에 적합한 것으로 확인되었다. 호프 추출물에 xanthohumol 표준용액을 5, 25, 50 μg/mL의 3개 수준으로 첨가하고 분석한 회수율은 96.76~102.93%로 나타나 높은 정확성이 있는 것을 확인할 수 있었다. 정밀성은 반복성과 재현성으로 확인했으며, 호프 추출물 125~500 mg 범위에서 시료량의 변화에 따른 반복성은 RSD 0.32~0.85%로 나타났고 실험자를 달리한 재현성 검증 결과 RSD는 0.18~ 0.70%로 확인되어 본 분석법은 정밀성이 있음을 확인하였다. 반응의 표준편차와 검량선의 기울기에 근거한 검출한계는 0.26 μg/mL였고 정량한계는 0.80 μg/mL로 호프 추출물의 xanthohumol 함량 분석을 위한 적합한 한계수준으로 확인되었다. 이상의 결과로 호프 추출물의 지표성분인 xanthohumol의 HPLC 분석법은 국내외 가이드라인에 적합한 시험법으로 검증되었다. 또한 검증된 분석법을 이용하여 호프 추출물의 xanthohumol 함량을 분석한 결과 6.40±0.75 mg/g으로 분석되었다. 본 시험법은 호프 추출물의 건강기능식품 개별인정원료 개발을 위한 표준화의 기초자료로 활용될 것으로 사료된다. This study was performed to establish a high-performance liquid chromatography (HPLC) analytical method for determining xanthohumol in hop (Humulus lupulus) extract for its standardization as a functional ingredient. The analytical method was validated based on the ‘Guideline on Preparing Dossiers for Functional Ingredient Recognition of Health Functional Food’ of the Ministry of Food and Drug Safety of Korea and the International Conference on Harmonization guidelines to determine its reliability and validity in terms of specificity, linearity, accuracy, precision, limit of detection (LOD) and limit of quantification (LOQ). The specificity was confirmed by the chromatogram obtained from the HPLC analysis method, as well as by the results of ultraviolet/visible spectra. The correlation coefficient (R2) was calculated to be close to 1, confirming that this was a suitable analysis with high linearity. Xanthohumol recovery ranged from 96.76 to 101.30% with relative standard deviation (RSD) values of less than 5%, and the repeatability and reproducibility of precision were confirmed to be 0.32∼0.85% and 0.18∼0.70% for the RSD, respectively, which implies that this method has high accuracy and precision. In addition, the LOD was 0.26 μg/mL, and the LOQ was 0.80 μg/mL, confirming that detection was valid at low concentrations. These results indicate that the proposed HPLC method for the detection of xanthohumol in hop extract is simple, reliable, and reproducible.

전시발표 : F 분야: 임산에너지 ; P-103 : 옥살산을 이용한 2단계 전처리의 반응 조건 변화를 통한 국산 소경목의 furfural 생산

류가희,정한섭,장수경,홍창영,최인규 韓國木材工學會 2015 한국목재공학회 학술발표논문집 Vol.2015 No.-

스피루리나 에탄올 추출물의 신경세포 보호활성

류가희,마충제 한국생약학회 2021 생약학회지 Vol.52 No.3

Excessive glutamate can cause oxidative stress in neuronal cells and this can be the reason for neurodegenerative disease. In this study, we investigated the protective effect of Spirulina maxima hot ethanol extract on mouse hippocampal HT22 cell of which glutamate receptor has no function. HT22 cells were pre-treated with S. maxima sample at a dose dependent manner (1, 10 and 100 µg/ml). After an hour, glutamate was treated. Cell viability, reactive oxygen species (ROS) accumulation, Ca2+ influx, decrease of mitochondrial membrane potential level and glutathione related assays were followed by then. S. maxima ethanol extract improved the cell viability by suppressing the ROS and Ca2+ formation, retaining the mitochondrial membrane potential level and protecting the activity of the antioxidant enzymes compared with group of vehicle-treated controls. These suggest that S. maxima may decelerate the neurodegeneration by attenuating neuronal damage and oxidative stress.

신갈나무 약산 전처리 공정 중 반응표면분석법을 이용한 액상 가수분해물의 furfural 수율 탐색

류가희 ( Ga Hee Ryu ),정한섭 ( Han Seob Jeong ),장수경 ( Soo Kyeong Jang ),홍창영 ( Chang Young Hong ),최준원 ( Joon Weon Choi ),최인규 ( In Gyu Choi ) 한국목재공학회 2016 목재공학 Vol.44 No.1

본 연구에서는 약산 전처리 공정을 통해 신갈나무의 액상 가수분해물로부터 유용 화합물인 furfural을 생산하였고 반응표면분석법을 이용하여 고수율의 furfural을 생산할 수 있는 최적 전처리 조건을 구명하였다. 전처리 공정은 반응 표면분석법 중 중심합성계획에 의하여 설계되었으며, 독립변수는 furfural 수율에 영향을 주는 반응온도(X1), 산 농도(X2), 반응시간(X3)으로 지정하였다. 종속변수(Y)는 약산 전처리로부터 생성된 furfural의 농도로 설정하였고 수율(Z)은 초기 시료 중량 대비 Y를 백분율로 나타내었다. 반응표면분석 결과, 반응온도(X1)가 furfural 수율에 가장 큰 영향을 주는 단일 독립변수로 나타났고, 두 변수의 영향을 보았을 때는 반응온도(X1)-산 농도(X2)의 상호작용이 furfural 수율에 가장 유의적인 인자로 확인되었다. 또한 반응표면분석법을 통해 예상된 약산 전처리의 최대 furfural 수율 조건은 반응온도(X1) 184℃, 산 농도(X2) 1.17%, 반응시간(X3) 5분이었으며 예상되는 최대 furfural 수율은 초기 시료 중량 대비 6.37%로 나타났다. 상기 최적 조건에서 실제 전처리를 수행한 결과, 생산된 furfural은 6.21%로 예상 수율과 근접하였으며 이를 통해 반응표면분석법을 이용하여 약산 전처리한 신갈나무의 액상 가수분해물로부터 생산된 furfural의 최적 수율 모델을 구축할 수 있었다. In this study, furfural, which is one of the value-added chemicals, was produced from the hydrolyzate of Quercus mongolica using dilute acid pretreatment, and the optimal pretreatment condition was determined by Response Surface Methodology (RSM) to obtain high yield of furfural. Based on Central Composite Design, the pretreatment experiment was designed with parameters such as reaction temperature (X1), acid concentration (X2), and reaction time (X3) as independent variables, while dependent variable was furfural concentration (Y), and furfural yield (Z) was shown as percentage of Y per a dry weight basis. According to results of RSM, it was confirmed that reaction temperature (X1) was the most influence factor and reaction temperature (X1)-acid concentration (X2) was the most significant interaction factor on furfural yield. Also, the optimal condition for the highest furfural yield was predicted at reaction temperature of 184℃, acid concentration of 1.17%, and reaction time of 5 min by RSM, and expected maximum yield of furfural was 6.37%. Experimentally, the maximum yield of furfural produced at above optimal condition was 6.21%, and it was considerably similar with the predicted value, and therefore the model for furfural production from the hydrolyzate of Quercus mongolica during dilute acid pretreatment could be built using RSM.

금전초에서 분리한 cynaroside의 신경세포보호 활성

마충제,류가희 한국생약학회 2023 생약학회지 Vol.54 No.1

In the previous study, we reported that cynaroside isolated from Lysimachia christinae methanolic extract had potent neuroprotective activities in neuronal cell death injured by excessive glutamate. In this study, we tried to confirm the neuroprotective activities of cynaroside in glutamate injured HT22 cells and establish mechanisms of neuroprotective action of cynaroside. We employed HT22 cells damaged by glutamate-induced cell death as a bioassay system. Cynaroside decreased reactive oxygen species increased by excessive glutamate treatment in HT22 cells. Also, Ca2+ concentration was decreased by cynaroside treatment. Cynaroside restored mitochondrial membrane potential to normal condition. It also increased not only glutathione reductase but also peroxidase to the control level. And it increased amount of glutathione, an endogenous antioxidant. These results suggested that cynaroside isolated from L. christinae showed potent neuroprotective activity through the anti-oxidative pathway.

Quantitative Analysis of Chemical Components of Hydrolysate from Silkworm Fed with Cudrania tricuspidata Leaves

최재환,남민지,류가희,전정욱,윤성섭 대한의생명과학회 2022 Biomedical Science Letters Vol.28 No.4

The purpose of this study was to develop hydrolysate from silkworm (Cudrania Silkworm Fibroin Peptide; CSFP), a food containing components for improving blood vessel health. In general, Cudrania tricuspidata leaves contain about 5 times more rutin and 25 times more dihydroquercetin than mulberry leaves. 1-Deoxynojirimycine (1-DNJ), dihydroquercetin and rutin inhibit the activity of carbohydrate-digesting enzymes, inhibit blood lipid peroxidation, and regulate insulin secretion, which helps blood vessels to be healthy. When the diet-controlled silkworm was enzymatically hydrolyzed, it was confirmed that rutin content was about 8 times higher than that of the in general silkworm as a control. In the silkworm hydrolysate, CSFP, developed as a final food material, the active ingredients were 65 mg/kg for rutin, 3,328 mg/ kg for DNJ, 0.43 mg/kg for dihydroquercetin, and 82,624 mg/kg for total polyphenol, which was confirmed through LC-MS/MS analysis. In conclusion, it was found that silkworms fed with C. tricuspidata leaves as a diet had more active components that can help control blood sugar and improve blood vessel health than silkworms fed with mulberry leaves.

Adipose sirtuin 6 drives macrophage polarization toward M2 through IL-4 production and maintains systemic insulin sensitivity in mice and humans

송미영,김상훈,류가희,김미경,차혜나,박소영,황홍필,유희철,배은주,박병현 생화학분자생물학회 2019 Experimental and molecular medicine Vol.51 No.-

Adipose tissue inflammation is a reproducible feature of obesity and obesity-linked insulin resistance. Although sirtuin 6 (Sirt6) deficiency has previously been implicated in diet-induced obesity and systemic insulin resistance, the adipocyte-specific role of Sirt6 in the regulation of adipose tissue inflammation and systemic metabolic dysfunction in mice fed normal chow and in humans remains elusive. Here, using Adipoq-Cre-mediated adipocyte-specific Sirt6 knockout (aS6KO) mice, we explored whether adipocyte Sirt6 inhibits adipose tissue inflammation and its underlying mechanism. aS6KO mice fed normal chow gained more body weight and fat mass than wild-type mice and exhibited glucose intolerance and systemic insulin resistance. Measurement of plasma and tissue cytokines and flow cytometric analysis of adipose stromal vascular cells indicated a decrease in alternatively activated M2 macrophages in the adipose tissue of aS6KO mice. Mechanistically, Sirt6 regulated the expression of the canonical type 2 cytokine IL-4 by adipocytes in a cell autonomous manner, which in turn affects M2 macrophage polarization. Consistent with animal experimental data, the degree of obesity and insulin resistance demonstrated by the body mass index, fasting blood glucose and HbA1c correlated negatively with the expression of Sirt6 in human visceral fat tissues. Collectively, these results suggest that adipocyte Sirt6 regulates body weight gain and insulin sensitivity independent of diet, and the increased IL-4 production by Sirt6 and resultant M2 polarization of adipose tissue macrophages may attenuate proinflammatory responses in adipose tissue.

Two-step 약산 전처리 공정 조건에 따른 낙엽송의 6탄당 전환 산물 및 당화 수율 평가

장수경,정한섭,류가희,박세영,최준원,여환명,최인규 한국목재공학회 2015 한국목재공학회 학술발표논문집 Vol.2015 No.-

당 생산 효율과 액상가수분해산물 분석을 통한 소경 낙엽송의 탈구조화 평가

장수경,정한섭,류가희,최준원,여환명,최인규 한국목재공학회 2014 한국목재공학회 학술발표논문집 Vol.2014 No.-

신갈나무 소경목을 이용한 SPORL 전처리 중 발생되는 C5 sugar의 전환 특성

정한섭,장수경,류가희,김호용,최준원,여환명,최인규 한국목재공학회 2014 한국목재공학회 학술발표논문집 Vol.2014 No.-