• 대한약침학회지
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• 제15권4호
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• pp.52-65
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• 2012

Objectives: Toad venom, called Chan-Su, is a traditional Oriental medicine secreted from the auricular and the skin glands of the Bufo bufo gargarizanz Cantor or B. melanosticus Schneider and has been widely used in China, Korea and other parts of Asia for the treatment of pain, heart conditions, and cancer. We examined the concentrations of the main chemical constituents within a commercially available toad venom product and compared the levels for different extraction methods. Methods: Toad venom was extracted using either cold or hot water, ethanol (EtOH), methanol (MeOH), or ethyl acetate (EtOAc), was fractionated using precipitation or reflux, and was then analyzed using thin layer chromatography (TLC), high-performance liquid chromatography (HTLC), and liquid chroma-tography - mass spectrometry (LC-MS). Individual components were identified by comparisons of the retention times, the ultraviolet spectra, and mass spectras and differences in chemical constituents for different solvents and extraction methods are presented. Results: Components with authentic standards, including serotonin and bufodienolides (cinobufagen, bufalin, cinobufalin, and resibufogenin), were detected. The water extract of toad venom contained the greatest amount of serotonin ($75.7{\pm}0.1$ mg/g), but very small amounts of bufodienolides ($3.8{\pm}0.0$ mg/g). In contrast, the use of MeOH or EtOH extraction solutions resulted in 5-26 times higher concentrations of bufodienolides, with only trace amounts of serotonin. The relative and the absolute concentrations of the component also varied based on the extraction method; i.e., EtOH extracts yielded the greatest total amounts of bufodienolides, and EtOAc precipitation had the lowest amounts of bufodienolides. Conclusions: Toad venom consists of serotonin and several bufodienolides, and the choice of solvent to extract chemical the constituents is important as a way to enrich the purported active components for treating different conditions.

• 약학회지
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• 제40권2호
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• pp.230-237
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• 1996

The fractions of Epimedii Herba were examined for the immunological effects in ICR mice. Mice were divided into 4 groups and administered orally the fractions of Epimedii Herba for 10 days. The results of this study were summarized as following: (1) The fraction 1 (EtOAc layer) administered group as compared with control group significantly decreased spleen weight, Arthus reaction and hemagglutination (HA) titer but significantly increased circulating white blood cells (WBC). (2) The fraction 2 ($H_20$ layer) administered group as compared with control group significantly decreased liver weight, Arthus reaction and HA titer but significantly increased WBC. (3) The fraction 3 (ppt) administered group as compared with control group significantly increased liver weight, thymus weight rate, delayed type hypersensitivity, phagocytic activity and WBC. The results showed that Frs. 1 and 2 administered groups reduced humoral immune response but increased WBC, and that Fr. 3 administered group increased cell-mediated immune response, phagocytic activity and WBC.

• 생약학회지
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• 제29권3호
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• pp.209-216
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• 1998

In the course of phytochemical studies of Rumex acetosella L. (Polygonaceae), the MeOH extract of the whole plants was fractionated with $CH_2Cl_2$ and $H_2O$, and the $CH_2Cl_2$ layer was fractionated again with 90% MeOH and hexane. Whereas $H_2O$ layer was fractionated with EtOAc and then with n-BuOH again. By repeated column chromatography, compound 1 was isolated from the 90% MeOH fraction, compounds 1, 2, 3, 4 and 5 were from the ethylacetate fraction, and compound 5 was from the n--BuOH fraction. On the basis of physico-chemical, spectroscopic evidences and in comparison with authentic samples, the structures of them were confirmed to emodin, citreorosein, $chrysophanol-8-O-{\beta}-D-glucopyranoside$, luteolin and $luteolin-7-O {\beta}-D-glucopyranoside$.

• 대한화장품학회지
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• 제39권2호
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• pp.159-166
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• 2013

본 연구에서는 새로운 미백소재를 개발하기 위해 적양 에탄올추출물을 효소처리 후 EtOAc 분획물(AJE)을 준비하여 in vitro 상에서 이들의 tyrosinase 저해활성과 세포 수준에서의 멜라닌 합성 저해효과를 측정하였다. AJE는 mushroom tyrosinase의 활성에는 영향을 미치지 않았으나 B16-F1 melanoma cell을 이용한 멜라닌 합성 저해효과에 있어서 농도 의존적으로 멜라닌 합성을 저해하여, $40{\mu}g/mL$의 농도에서 52% 이상의 저해효과를 나타내었다. 이러한 멜라닌 합성 저해효과에 대한 작용 기전을 확인하기 위해 western blot을 통해 멜라닌 합성 경로에 관련된 단백질의 발현을 측정하였다. 그 결과 멜라닌 합성에 관여하는 효소인 tyrosinase related protein 1 (TRP-1)의 발현을 억제하였고, 이를 조절하는 전사인자인 microphthalmia associated transcription factor (MITF) 발현 역시 효과적으로 억제하였다. 또한 extracellular signal-regulated kinase (ERK) pathway를 활성화시킴으로써 phosphorylated extracellular signal-regulated kinase (p-ERK)의 발현을 상당히 증가시키는 것을 확인할 수 있었다. 이러한 결과는 AJE가 멜라닌 합성의 신호전달 경로 중 ERK pathway의 활성화를 통해 MITF의 분해를 촉진시키고 이로 인해 MITF의 발현을 감소시키며, 그 결과 멜라닌 합성에 관여하는 효소 중 TRP-1의 발현을 감소시킴으로써 멜라닌 합성을 저해하는 것으로 사료되며, 따라서 AJE는 미백용도의 기능성 원료로서의 가능성이 큰 것으로 판단된다.

• 한국작물학회지
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• 제46권6호
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• pp.449-452
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• 2001

Legume seed isoflavones may have a variety of desirable physiological effect on the human health including both the circulatory and skeletal systems. The present study was performed to determine the isoflavone content of leaf and seed as well as to purify and identify the types of isoflavone from leaf extract of hyacinth bean (Lablab purpureus (L.) Sweet). Reverse phase HPLC revealed six different types of isoflavone such as daidzin, genistin, daidzein, genistein, 6"-o-acetyl genistin and 6"-o-acetyl daidzin in aqueous methanol extract from seeds and leaves of the hyacinth bean. Relatively, leaf isoflavone content of hyacinth bean was greater than seed isoflavone content. Using DiAion HP-20 silica gel and sephadex LH-20 chromatography, pure daidzein was identified in the ether layer, whereas genistin was in the EtOAC fraction.

• 한국식품위생안전성학회지
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• 제19권3호
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• pp.167-170
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• 2004

The present study was undertaken to investigate the effects of Moutan Cortex Radicis extracts and paeonol, a major component, on rabbit platelet aggregation and thromboxane (TX) $A_2$ formation. Moutan Cortex Radicis methanol and butanol layers (100 ${\mu}g/mL$) showed the weak inhibitions, and water layer (100 ${\mu}g/mL$) had no effect on the collagen-induced platelet aggregation. Whereas, hexane and EtOAc layers potently inhibited the collagen (3 ${\mu}g/mL$)-induced platelet aggregation with the $IC_{50}$ values of 10.9${\pm}$1.0 and 31.5${\pm}$0.8 ${\mu}g/mL$, respectively. Paeonol isolated from the hexane-acetone layer specifically inhibited the collagen-induced platelet aggregation with the $IC_{50}$ value of 113.1 ${\pm}$ 0.9 ${\mu}M$, whereas it had little effects on the other agonists such as AA-, thrombin-, A23187- and thapsigargin-induced platelet aggregations. Paeonol also potently inhibited the collagen-induced TXB formation in rabbit platelet in a concentration-dependent manner. These results suggest that paeonol may inhibit rabbit platelet aggregation by interfering with an essential step in collagen-induced platelet activation and $TXA_2$ formation. Paeonol may be a promising candidate for an antiplatelet agent.

• 한국자원식물학회지
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• 제19권4호
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• pp.491-496
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• 2006

18종의 한약재 MeOH 추출물과 EtOH 추출물의 Staphylococcus aureus에 대한 항균력을 조사한 결과 하고초, 소목, 오배자 및 향유는 5mg/ml 수준에서 항균활성을 나타내었으며 지실, 금은화 및 황금 등은 10mg/ml, 오미자, 초두구 및 희렴 등은 30mg/ml 이상에서 항균성을 보였다. 최소저해농도는 MeOH추출물에서는 $1{\sim}34mg/ml$, EtOH추출물에서는 $1{\sim}46mg/ml$ 농도에서 항균활성을 보였으며 분획 층 중 EtOAc층에서 성장억제효과가 가장 높게 나타났다. 항균력이 가장 우수한 것으로 확인된 소목, 오배자 및 황금추출물의 미생물 증식억제 효과를 조사하기 위해 증식배지에 각각의 추출물을 0, 100, 300 및 500ppm의 농도로 첨가하여 균주의 증식을 조사한 결과 배양후 24시간에 소목 추출물 무첨가구의 $OD_{620}$값이 0.4인 반면 100ppm 이상의 추출물 첨가 시 $0.02{\sim}0.03$정도로 균증식이 현저히 억제되었고, 오배자 추출물 무첨가구의 $OD_{620}$값이 0.4인 반면 300ppm 이상의 추출물 첨가 시 $0.02{\sim}0.03$정도로 균증식이 현저히 억제되었으며 황금 추출물 무첨가구의 $OD_{620}$값이 0.4인 반면 300ppm 이상의 추출물 첨가 시 $0.02{\sim}0.07$ 정도로 균증식이 현저히 억제되었다. 이상의 결과로 볼 때 제대염, 부종성 피부염 및 관절염 등의 다양한 가축질병을 일으키는 Staphylococcus aureusi의 감염을 예방하거나 또는 치료할 수 있는 한약재 추출물 사료첨가제의 개발 가능성을 확인 할 수 있었다.

• 한국식품과학회지
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• 제40권2호
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• pp.129-134
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• 2008

복분자 와인은 복분자 열매를 발효ㆍ숙성시켜 제조하기 때문에 그들 양자 간의 성분에는 차이가 있을 것으로 추측되었다. 그러나 복분자 와인의 음용률이 높음에도 불구하고 복분자 와인 중에 함유된 성분들에 관한 체계적인 연구는 거의 전무한 실정이다. 그래서 먼저 복분자 와인 중에 함유되어 있는 성분들의 chemical profile의 검토를 위해 항산화 활성에 초점을 맞추어 화합물의 단리ㆍ구조해석에 착수하였다. 복분자 와인 11 L(신선 복분자 열매 15.7 kg 상당량)의 농축액(928.3 g)을 용매분획한 후 얻어진 EtOAc층(20 g/56.2 g)을 대상으로 각종 column chromatography 및 HPLC를 행하여 5종의 항산화 활성 화합물을 단리하였다. 그들을 대상으로 $^1H$-NMR 분석 및 TMS 유도체화 후 GCMS 분석을 행한 결과, 4-hydroxybenzoic acid(1, 0.1 mg), 3,4-dihydroxybenzoic acid(2, 0.3 mg), 4-(2-hydroxyethyl)-phenol(3, 0.6 mg), pyrocatechol(4, 0.3 mg), 3,4,5-trihydroxybenzoic acid ethyl ester(5, 0.6 mg)로 동정하였다. 화합물 1과 2는 복분자 열매중에 존재함이 보고되어 있으나 화합물 3-5는 복분자 열매로부터 그 존재가 보고되어진 바 없으며, 이들 5종의 화합물 모두가 복분자 와인으로부터 동정되어진 것 또한 처음이다.

• 생약학회지
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• 제39권4호
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• pp.347-351
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• 2008

The methanol (MeOH) extract of the rhizome of Alpinia officinarum Hance (Zingiberaceae) demonstrated a potent inhibition on the proliferation of cultured human tumor cell lines such as MES-SA (human uterine carcinoma cell line), MESSA/DX5 (multidrug resistant subline of MES-SA), HCT-15 (human colorectal adenocarcinoma cell line), HCT15/CL02 (multidrug resistant subline of HCT15). The MeOH extract was fractionated into four portions by serial solvent partition, ie., methylene chloride (CH2Cl2) soluble part, ethylacetate (EtOAc) soluble part, n-butanol (BuOH) soluble part and remaining water layer. Among them, the $CH_2Cl_2$ soluble part of the extract exhibited a most potent inhibition on the proliferation of tested tumor cell lines. Bioassay-guided fractionation of the $CH_2Cl_2$ soluble part led to the isolation of five diarylheptanoid and two flavonoid constituents, i. e., galangin (1), 7-(4"-hydroxy-3"-methoxyphenyl)-1-phenylhept-4-en-3-one (2), 1,7-diphenyl-5-hydroxy-3-heptanone (3), trans,trans-1-(3'-methoxy-4'-hydroxyphenyl)-7-phenyl-5-ol-4,6-dien-3-heptanone (4), 5-methoxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (5), kaempferide (6), 5-hydroxy-7-(4"-hydroxy-3"-methoxyphenyl)-1-phenyl-3-heptanone (7). Structures of the isolated active components (1 - 7) were established by chemical and spectroscopic means.

• Natural Product Sciences
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• 제3권1호
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• pp.29-37
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• 1997

Protein Kinase C (PKC) is generally believed to play a central role in signal transduction, cellular growth control, gene expression, and tumor promotion. And it has been suggested that inhibitors of PKC might play important roles for the prevention and treatment of cancer. In order to investigate the possible inhibitors of PKC from natural products, PKC receptor binding assay was performed using bovine brain particulate as a source of PKC and the amount of $[^3H]Phorbol$ 12,13-dibutyrate (PDBu) bound to PKC was measured in the presence of test materials. Total methanol extracts from 100 kinds of natural products were partitioned into 3 fractions (n-hexane, ethyl acetate and aqueous layer) and their binding ability to the regulatory domain of PKC was evaluated. The ethyl acetate fractions of Morus alba $(roots,\;IC_{50}:\;156.6\;{\mu}g/ml)$, Rehmannia glutinosa $(roots,\;IC_{50}:\;134.3\;{\mu}g/ml)$, Lysimachia foenum-graecum $(roots,\;IC_{50}:\;167.8\;{\mu}g/ml)$, Polygonum cuspidata $(roots,\;IC_{50}:\;157.3\;{\mu}g/ml)$, Cnidium officinale $(aerial\;parts,\;IC_{50}:\;145.2\;{\mu}g/ml)$, and the hexane $(IC_{50}:\;179.3\;{\mu}g/ml)$ and the EtOAc fraction of Symplocarpus nipponicus $(roots,\;IC_{50}:\;155.9\;{\mu}g/ml)$ showed inhibitory activity of $[^3H]PDBu$ binding to PKC.