우슬의 뿌리로부터 Phytoecdysones의 분리와 항 아토피 효과

김기옥 ( Ki Ohk Kim ),구창섭 ( Chang Sub Ku ),김민진 ( Min Jin Kim ),박윤정 ( Yhun Jung Park ),류형원 ( Hyung Won Ryu ),송혁환 ( Hyuk Hwan Song ),김정희 ( Jung Hee Kim ),오세량 ( Sei Ryang Oh ) 한국응용생명화학회(구 한국농화학회) 2015 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.1

본 연구에서는 우슬 열수 추출물을 n-BuOH과 H2O로 용매 분획 한 후 n-BuOH 분획물을 컬럼 크로마토그래피를 수행하여 6개의 소분획물(Fr. 1-6)을 얻었다. 소분획물 6개에 대해 tumor necrosis factor (TNF)-α 저해활성 검정을 통하여 가장 높은 활성을 나타낸 분획물 5에서 preparative-high performance liquid chromatography를 이용해 3종의 phytoecdysone계열 화합물들을 분리하였다. 분리된 화합물들은 NMR 및 MS 분석을 통해 ecdysterone (1), 25S-inokosteron (2), 25R-inokosteron (3)임을 동정 하였고, 이들의 활성은 TNF-α 생산 저해활성은 200 μg/mL에서 80-95%의 TNF-α 저해활성을 나타내었다. 또한 각질형성 세포에 분리한 3종의 화합물을 처리한 결과 각각 51와 23% 이상의 TARC의 생성 억제 효과를 확인 할 수 있었다. 이들의 결과로써, 우슬 추출물과 분리된 화합물들에 대한 항 아토피 활성 결과로 천연물 기능성 식품 또는 화장품으로의 응용 가능성이 높다고 사료된다. The roots of Achyranthes japonica Nakai were extracted with 100% aqueous and concentrated subfraction was separated with ultra-performance liquid chromatography-based activity profiling. Three compounds were isolated from the subfraction 5 through the repeated prep- high performance liquid chromatography column chromatography. According to the results of physico-chemical and spectroscopic data including NMR and MS, the chemical structures of the compounds were determined as ecdysterone (1), 25S-inokosterone (2), and 25R-inokosterone (3). Three phytoecdysones were showed weak inhibitory activity for thymus and activation-regulated chemokine expression levels in tumor necrosis factor (TNF)-α plus IFN-γ induced HaCaT cells, respectively. However, those compounds 1-3 were exhibited the most potent inhibition (80-95% at 200 μg/mL) against TNF-α expression levels in A23187 plus phorbol-myrisrate acetate-induced RBL-2H3 cells. As result, 100% aqueous extract of A. japonica has an excellent anti-atopy activity. It could be used to a large range of functional anti-atopy cosmetics.

미세먼지 자극으로부터 지모추출물, 베타글루칸, 폴리감마글루탐산 혼합물의 안티폴루션 효능 평가

서수경 ( Soo-kyung Seo ),구창섭 ( Chang-sub Ku ),손보미 ( Bomi Son ),이수경 ( Su-gyeong Lee ),강은미 ( Eun Mee Kang ),김미옥 ( Miok Kim ),홍정현 ( Jung Hyun Hong ),김선화 ( Seonhwa Kim ),신문삼 ( Moon Sam Shin ) 대한미용학회(구 대한미용과학회) 2016 대한미용학회지 Vol.12 No.4

This study characterized Prostaglandin E2 (PGE2) production from Raw264.7 cells stimulated with particulate matter (PM1648a) and tested anti-pollution capacity of mixtures of Anemarrhena asphodeloides root extract, β-glucan and poly- γ-glutamic acid (the complex). In order to assess cytotoxicity, we treated the Raw264.7 cells with the complex using the lactase dehydrogenase (LDH) assay method. To measure cell viability, we treated HaCaT cells with the complex using the MTT assay method. To test scavenging activity, the DPPH assay method was used. To test the anti-oxidant effect of the complex, we treated HaCaT cells with the pro-inflammatory mediator particulate matter (PM1648a) and with PM1648a and the complex, then measured the concentration of reactive oxygen species (ROS) within the cells using the dichlorofluorescin diacetate (DCF-DA) assay method. The complex had cytotoxicity below 6.5% and cell viability above 91.3% at a concentration of 0.05%. Also, the complex showed scavenging activity above 50% at a concentration of 1.0%. In addition, ROS production at the 0.05% complex concentration was reduced to 61.0%. In a clinical experiment, the subjects who had skin urtication used a cosmetic product containing mixtures of 1.0% Anemarrhena asphodeloides root extract, 0.4% β-glucan and 0.5% poly-γ-glutamic acid for 4 weeks. The experimental group showed a statistically significant improvement in skin humidity, desquamation index and skin urtication. Based on the results with in vitro test and clinical experiment, mixtures of Anemarrhena asphodeloides root extract, β-glucan and poly-γ-glutamic acid could be used as cosmetic anti-pollution materials.

지모의 뿌리줄기로부터 분리된 nyasol의 미용효과

박윤정 ( Yhun Jung Park ),구창섭 ( Chang Sub Ku ),김민진 ( Min Jin Kim ),이미경 ( Mi Kyeong Lee ),김기옥 ( Ki Ohk Kim ),류형원 ( Hyung Won Ryu ),송혁환 ( Hyuk Hwan Song ),김두영 ( Doo Young Kim ),오세량 ( Sei Ryang Oh ) 한국응용생명화학회(구 한국농화학회) 2015 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.1

지모 뿌리를 100% EtOH로 추출, 농축하여 medium pressure liquid chromatography 기반의 NO 분획물 활성 검정법을 이용하여 활성을 검정하였다. preparative high performance liquid chromatography를 반복적인 칼럼크로마토그래피를 수행하여 1개의 단일성분을 분리하였다. NMR, MS 등을 포함한 이화학 및 분광학적 자료에 근거하여 분리된 물질을 구조 분석한 결과 (-)-nyasol로 구조동정 하였다. 분리된 활성물질 nyasol은 NO 생성 억제, melanin과 tyrosinase를 억제하는 미백효과, thymus and activation-regulated chemokine 생성을 억제하는 활성에서 6.25-12.5 μg/mL의 IC50 값을 가지는 강한 저해 활성을 나타냈다. 특히 nyasol은 염증에 의한 항아토피 및 항미백활성에 있어 뛰어난 효능을 나타내었다. 이 결과로 보아 nyasol은 천연 기능성 화장품으로써 이용 가능성을 넓힐 수 있을 것으로 사료된다. The rhizomes of Anemarrhena asphodeloide was extracted with 100% ethanol and concentrated subfractions were separated with medium pressure liquid chromatography-based activity profiling. One compound was isolated from the subfraction 10 through the repeated preparative high performance liquid chromatography (prep-HPLC). According to physico-chemical and spectroscopic data including NMR and MS, the chemical structures of the compound was determined as nyasol (1). Nyasol was exhibited potent inhibitory activity for NO (IC50: 12.5 μg/mL), tyrosinase (IC50: 12.5 μg/mL), melanin contents (IC50: 12.5 μg/mL), thymus and activation-regulated chemokine (TARC) production (IC50: 6.25 μg/mL). As a result, nyasol has an excellent inflammation-dependent anti-whitening and TARC production activity. It could be used to a large range of functional cosmetics.

연잎 추출물을 함유한 화장품의 안정성 평가

최선주(Sun Ju Choi),김소영(Soyoung Kim),정윤주(Yoonjoo Jeong),구창섭(Chang Sub Ku),하병집(Buungjhip Ha),채희정(Hee Jeong Chae) 한국생물공학회 2011 KSBB Journal Vol.26 No.1

The stability of cream containing lotus leaf extract (LLE) was evaluated. Stability pH and viscosity were measured at 3 different temperatures (25℃, 45℃ and 65℃), and at room temperature under the sun light at 2 day intervals during 12 days. The cleansing cream and foam cleansing containing the extract did not show a significant pH change at the different temperatures. However, the pH of the cleansing cream and foam cleansing containing LLE slightly decreased at 25℃. They showed a high stability at temperature conditions (45℃ and 65℃) and under the sun light condition. The cleansing cream and foam cleansing containing LLE did not show a significant viscosity change at 25℃, but had a tendency of decrease at 45℃ and 65℃. There was no significant offflavor and discoloration as well as physical changes such as creaming and cohesion at 25-65℃ and under the sun light condition. And this LLE could be used as a stable functional cosmetic material.

사과꽃잎 추출물의 용매 분획으로부터 기능성 소재의 탐색

최선주(Sun Ju Choi),조은아(Eun-ah Cho),조은혜(Eunhye Cho),정윤주(Yoonjoo Jeong),구창섭(Chang Sub Ku),하병집(Byungjhip Ha),채희정(Hee Jeong Chae) 한국생물공학회 2011 KSBB Journal Vol.26 No.2

Fractional solvent extraction by organic solvents such as hexane, chloroform, ethylacetate, and butanol was carried out using 70% ethanol extract of apple flower leaves. Biological activities including antioxidant, whitening, antimicrobial and anti-wrinkle activities were investigated and bio-active materials of the extracts were identified using GC/MSD. Among the tested solvent fractions, ethylacetate fraction showed the highest total polyphenol content (1218.94 μg/mL), and flavonoid (140 μg/mL). The DPPH radical scavenging activities was over 80% at a dry matterbased concentration of 200 μg/μL and SOD-like activity was over 90% at 50 μg/mL concentration in ethylacetate fraction that was slightly lower than of ascorbic aicd. Tyrosinase inhibition activity related to skin-whitening was over 60% by ethylacetate fraction of 100 μg/mL. As an anti-aging effect, elastase inhibitory activity was about 45% in ethylacetate fraction. Also, it showed a significantly antimicrobial activity against P. acenes. From GC/MSD analysis, a characteristic peak of high content in ethylacetate fraction was identified as kaempferol, which has been reported as a bioactive compound.

반하(Pinellia ternata)에서의 [6]-gingerol 함량과 멜라닌 저해 활성에 영향을 미치는 원산지 판별 마커로의 활용

류형원 ( Hyung Won Ryu ),안주현 ( Ju Hyeon An ),원효준 ( Hyo Jun Won ),서수경 ( Soo-kyung Seo ),김두영 ( Doo-young Kim ),구창섭 ( Chang-sub Ku ),오세량 ( Sei-ryang Oh ) 한국응용생명화학회(구 한국농화학회) 2016 Journal of Applied Biological Chemistry (J. Appl. Vol.59 No.4

반하(Pinellia ternata Breitenbach)는 천남성과(Araceae)의 약용식물로 동아시아 일대가 원산지이며, 유럽과 북아메리카 일부지역에도 널리 분포하고 있는 식물이다. 반하의 괴경(tuber)은 약재로 사용되어 구토, 염증, 외상을 치료하는 목적으로 사용되고 있다. 보고된 약리학적 작용으로는 항경련, 항종양, 살충, 세포독성 등이 있는 것으로 알려져 있다. 반하는 약리학적으로 유용한 약용식물로 잘 알려져 있음에도 불구하고, 원산지를 판별할 수 있는 신뢰성이 있고 표준화된 방법이 없는 실정이다. 이를 위한 한국산과 중국산 반하를 판별할 수 있는 유의적인 화합물을 탐색하기 위해 UPLC-PDA와 QTof-MS에 기반을 둔 대사체 표지법을 이용하였다. 한국산으로부터 원산지 판별 화합물로 예상되는 화합물을 반복적인 역상 칼럼크로마토그래피에 기반을 둔 활성유도 분획법을 통해 분리하였다. 그리고, NMR과 MS를 포함한 물리화학적, 분광학적 정보의 결과를 토대로 [6]- gingerol이라는 화합물의 구조를 동정하였다. [6]-gingerol은 원산지를 판별할 수 있는 능력과 melanin 생합성 저해 활성을 지닌 기능성 화장품 소재로서 가치가 있다고 판단되어짐에 따라 유효성분 원산지 판별 화합물로 Fingerprint법에 의해 선정되었다. 더욱이, 한국산과 중국산 반하 유래 [6]-gingerol의 함량 비교를 위해, 유효성이 검증된 분석법을 이용하여 이에 대한 검량곡선을 작성하여 그 함량을 비교하였다. 이것은 한국산 반하판별을 위한 화합물 선정과 성공적인 유효성 검증을 다룬 최초의 보고이다. Pinellia ternata Breitenbach, the natural medicinal plant of the Araceae family, is a perennial plant originated from the East Asia, but also widely distributed in Europe and North America. Its tuber is used as traditional medicine for treatment of various diseases such as vomiting, inflammation, and traumatic injury. Pharmacological studies revealed that P. ternata possesses anticonvulsant, anti-tumor, insecticidal, and cytotoxic activities. Despite being well-known as the useful medicinal plant, there is no reliable, standardized method for origin discrimination. Ultra performance liquid chromatography-photodiode array detector and quadrupole time of flight-mass spectrometry based metaboliteprofiling was applied to explore significant metabolite for origin discrimination between Korean and Chinese P. ternata. One compound was isolated from Korean P. ternata using repeated ODS column chromatography by bioactivity guided fractionation, and determined as [6]-gingerol according to the results of spectroscopic data including nuclear magnetic resonance and MS. This compound was selected as cosmeceutical biomarker by fingerprints, and it was associated to melanin inhibitory effect determining its origin authenticity. Furthermore, the calibration curve of biomarker was prepared using validated method for the comparison of content between Korean and Chinese P. ternata. This is the report to address the selection and successful validation of the discriminant metabolite for confirmation of Korean P. ternata.

장기간 숙성에 따른 죽초 및 목초액의 이화학적 변화

구창섭,문성필,박상범,권수덕 한국목재공학회 2002 목재공학 Vol.30 No.4

졸참나무, 맹종죽 및 소나무로부터 제조한 미정제 죽초 및 목초액을 약 1년 간 저장하고, 이들의 이화학적 변화를 주기적으로 분석하였다. pH, 유기산·용해타르 함량 및 비중은 전 숙성기간 중 큰 변화가 관찰되지 않았다. 그러나 색차의 경우 7∼10개월 사이에서 현저한 변화를 나타내었다. 또한 이 기간동안에 이들 죽초 및 목초액의 색깔은 연한 노랑 띤 오렌지색에서 자주색과 오렌지색으로 크게 변화하였다. 죽초 및 목초액 중 유기산 및 중성 성분(dihydro-2(3H)-furanone, furfural, furfuryl alcohol, 2-hydroxy-1-methyl-1-cyclopenten-3-one, 1-hydroxy-2-propanone, methanol)들의 함량은 3개월 간격으로 주기적인 증감을 나타내었다. 특히 유기산류와 중성 성분들의 관계는 0.92 정도의 높은 상관관계를 나타내었다. 페놀류의 함량은 비록 유사한 주기적인 증감이 관찰되었으나, 그 함량의 경우 숙성 전 기간에 걸쳐 감소하였다. Three different kinds of the crude vinegars obtained from oak(Quercus serrata), bamboo(Phyllostachys pubescens) and pine(Pinus densiflora) species were stored for approximately one year and periodically analyzed to monitor their physicochemical changes. Small changes in physical properties, such as the pH, specific gravity and amount of organic acids as well as water-soluble tar were observed in the entire course of aging period. However, the color difference of the vinegars showed a remarkable change between 7 and 10 months. In addition, these vinegars' colors changed from light yellowish orange to much deeper purple and orange during this period. The amount of organic acids and neutral compounds(dihydro-2(3H)-furanone, furfural, furfuryl alcohol, 2-hydroxy-1-methyl-1-cyclopenten-3-one, 1-hydroxy-2-propanone and methanol) in the vinegars increased or decreased periodically every three months. A good linear relationship (correlation coefficient of ca. 0.92) was obtained between the amount of organic acids and the amount of neutral compounds in such changes. However, although the amount of phenols increased or decreased periodically, its amount was decreased over the entire aging period.

3종류의 탄화로에서 얻어진 죽초액의 특성

구창섭,문성필,박상범,권수덕 한국목재공학회 2002 목재공학 Vol.30 No.4

3종류의 국내산 대나무의 생재 및 기건재를 3종류의 탄화로에서 탄화시키고 얻어진 죽초액에 대하여 그 특성을 검토하였다. 기계식 전용탄화로의 경우, 대부분 1단 회수 죽초액의 비중, 유기산 및 용해 타르 함량이 높았으나 2단 이상에서 감소하여 3단 이후에 일정해지는 경향을 나타내었으며 전체적으로는 기건재가 생재에 비해 이들 물성이 높게 나타났다. pH의 경우 맹종죽 기건재를 제오하고 생재가 기건재보다 높게 나타났다. 3성분합계는 비중, 유기산, 용해타르 함량과 유사한 경향을 나타내었다. 개량탄화로의 경우, 수종에 따라 전체 물성이 다양하게 나타났다. 간이탄화로에서 탄화시켜 연통온도 80℃로 얻어진 기건 죽초액의 경우 그 이상의 온도에서 얻어진 죽초액보다 비중, 용해타르 함량 및 3성분 합계가 높게 나타났다. 간이, 개량 및 기계식 전용탄화로에서 얻어진 죽초액의 비중은 온도, 수분함유 상태에 상관없이 유기산과 용해타르 함량의 합계와 0.90 정도의 높은 상관관계를 나타내어 앞으로 죽초액 품질 결정에 중요한 지표로 사용될 가능성이 시사되었다. 죽초액의 구성 유기성분의 경우 초산이 주성분이었으며, 리그닌의 열분해에 의하여 생성되는 guaiacyl 및 syringyl골격의 페놀류가 다량 검출되었다. Three different species of green and air-dried Korean bamboos were carbonized by using three different types of kilns designated as special (800∼1000℃), improved(600∼700℃) and simple kiln (400∼500℃), and the bamboo vinegars obtained from the carbonization processes were characterized. In the case of the special kiln most of the bamboo vinegars obtained at the first recovery stage showed high values of specific gravity and also in content of organic acid and water-soluble tar. The bamboo vinegars obtained from the improved kiln showed various physical properties depending on their species. In the case of simple kiln, the bamboo vinegars obtained from air-dried bamboos and at temperatures below 80℃, showed a higher specific gravity and more water-soluble tar as well as total organic components than those obtained at 80∼150℃. A good linear relationship (correlation coefficient of ca. 0.90) was obtained between the specific gravities and the sum of organic acids and water-soluble tars. Therefore, this correlation coefficient might be a good index to determine the quality of bamboo vinegars. The major chemical constituents of the bamboo vinegars were acetic acid and considerable amounts of phenols: guaiacol, ethyl guaiacol, syringol, and methyl syringol.

SPME법에 의한 죽초 및 목초액 중의 휘발성 성분 분석

문성필,구창섭 한국목재공학회 2002 목재공학 Vol.30 No.4

졸참나무 (Quercus serrata), 맹종죽 (Phyllostachys pubescens) 및 소나무 (Pinus densiflora)로부터 제조한 미정제 초액의 휘발성 성분을 고상(固相) 미량추출(solid phase microextraction; SPME)법을 이용하여 분석하였다. 이들 초액 중의 휘발성화합물 분석을 위하여 극성 (CBP 20) 및 무극성 (CBP 1) 칼람을 사용하였으며, 이로부터 총 264개의 피크를 검출하였다. 이들 성분들 중 주요 화합물은 2-butanone, acetic acid, guaiacol, phenol, 4-ethyl guaiacol, cresol류, 4-ethyl phenol, 그리고 syringol이었다. 무극성 칼람을 사용함에 의하여 7개의 성분, 즉, 1-hydroxy-2-butanone, ethisolide, furfuryl acetate, 1-(2-furanyl)-1-propane, 1,2-dimethoxybenzene, 1,2-dimethoxybenzyl alcohol, phenyl acetate를 새로이 동정할 수 있었다. 이 성분들 중 페놀류가 주성분이었으며, 휘발성 성분의 49∼65%를 차지하였다. 죽초액의 경우 페놀류의 비율은 다른 두 목초액보다 낮았다. 그러나 중성화합물류 및 유기산류는 소나무 및 졸참나무로부터 제조한 목초액보다 그 비율이 더 높았다. 따라서 이들 죽초액과 목초액 간의 훈취의 차이는 목초액의 서로 다른 휘발성분의 함유량의 차이에 기인한다고 생각된다. Volatile compounds in three different kinds of crude vinegars obtained from oak (Quercus serrata), bamboo (Phyllostachys pubescens) and pine (Pinus densiflora) species were analyzed by the solid-phase microextraction (SPME) method. A total of 264 peaks were detected on the chromatograms obtained from the polar (CBP 20) and the nonpolar (CBP 1) columns, which were used for analyzing the volatile compounds in these vinegars. The major volatile compounds identified by using the polar column were 2-butanone, acetic acid, guaiacol, phenol, cresols, 4-ethyl guaiacol, 4-ethyl phenol, and syringol. Using the nonpolar column, seven compounds could be identified: 1,2-dimethoxybenzyl alcohol, 1-hydroxy-2-butanone, 1-(2-furanyl)-1-propane, ethisolide, furfuryl acetate, 1,2-dimethoxybenzene, phenyl acetate. The volatile compounds were classified into five groups: phenols, neutral compounds, organic acids, esters and others. The phenols were the main component and comprised 49∼65% of the volatile compounds of these vinegars. In the case of bamboo vinegar, the proportion of the phenols in the volatile compounds was lower than that of the two wood vinegars. However, the proportions of the neutral compounds and the organic acids were higher than those of the wood vinegars. Therefore, it seems that these differences of the proportions of the volatile compounds would make a certain difference of a smoke flavor between the bamboo vinegar and the wood vinegars.