• Archives of Pharmacal Research
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• 제26권11호
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• pp.929-936
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• 2003

Methanol extract (MeOH), n-hexane (Hx), chloroform ($CHCl_3$), ethyl acetate (EA), butanol (BuOH) and aqueous ($H_2O$) fractions of Eucommiae Cortex including geniposidic acid (GA), geniposide (GP) and aucubin (AU) were tested for their therapeutic efficacy on osteoporosis. The contents of GA, GP and AU in the cortex and leaf of Eucommia ulmoides Oliver were quantified by HPLC. The effect of Eucommiae Cortex on the induction of growth hormone (GH) release was studied by using rat pituitary cells. The proliferation of osteoblast-like cells increased by herbal extracts was assayed using a tetrazolium (MTT), alkaline phosphatase (ALP) activity, and [$^3H$]-proline incorporation assays. The inhibition of osteoclast was studied by using the coculture of mouse bone marrow cells and ST-2 cells. As a result, the GA, GP and AU were present in the cortex more than in the leaf of E. ulmoides Oliver. The MeOH (1mg/mL), Hx, $CHCl_3$ and EA fractions (each 20 $\mu$ g/mL) had potent induction of GH release. The $CHCl_3$ exhibited the potent proliferation of osteoblasts. The AU, GP and GA were increased proliferation of osteoblasts. In addition, GA ($IC_{50}: 4.43{\times}10^{-7}$M), AU and GP were significantly inhibited proliferation of osteoclast. In summary, it is thought that the components in a part of the fractions of Eucommiae Cortex participate in each step of mechanism for activating osteoblast to facilitate osteogenesis, and suppress osteoclast activity to inhibit osteolysis.

• 한국동물위생학회지
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• 제27권1호
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• pp.17-29
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• 2004

A multiresidual analysis was performed to determine 12 sulfonamides(sulfacetamide, sulfadiazine, sulfisomidine, sulfathiazole, sulfapyridine, sulfamerazine, sulfamethazine, sulfamonomethoxine, sulfisoxazole, sulfamethoxazole, sulfaquinoxaline, and sulfadimethoxine) in beef and pork simultaneously. The multiresidual analysis for the sulfonamides currently used was able to analyze 5 kinds of sulfonamides at the same time. The method of this 12 sulfonamides multiresidual analysis in this study was matrix solid-phase dispersion(MSPD) by high performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS). The recovery rate of the materials was measured by MSPD method with 3 different extraction solvents; Dichloromethane, DCM: Ethylacetate(3:1), DCM:EA(9:1). Also, samples (84 beef and 205 pork samples) which were positive by EEC-4 plate test from 2001 to 2003 were tested to investigate the kinds of sulfonamides using HPLC. The results from the study were as follows; 1. The recovery rate of the materials was measured by MSPD method with 3 different extraction solvents; Dichloromethane, DCM:Ethylacetate(3:1), DCM:EA(9:1). The method of extraction solvent with DCM:ethyl acetate(9:1) was the most excellent(87.7∼99.3%) in separation and reappearance. 2. In the LC/MS analysis. of sulfonamides, signal to noise ratio was showed relatively high in the positive mode and special ion in the quality analysis was determined via [M+H]$\^$+/ and m/z 156. A spectrum of sulfonamides was showed from all 12 sulfonamides. 3. The samples positive by the EEC-4 plate, a screening test method, were categorized by sulfonamides through Charm II and confirmed the kinds of sulfonamides through HPLC. 1) Among 84 beef samples positive by EEC-4 plate, 20 samples were positive by Charm II and identified as 7 sulfamethazine, 9 sulfadimethoxine, 1 sulfamonomethoxine and 3 unknown status. 2) Among 205 pork samples positive by EEC-4 plate, 42 samples were positive by Charm II and identified as 19 sulfamethazine, 1 sulfadimethoxine, 4 sulfamonomethoxine and 5 unknown status.

• 대한한의학회지
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• 제32권1호
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• pp.141-150
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• 2011

Objectives: The aim of this study was to elucidate the antioxidant effect of solvent fractions of Angelica gigas root. Methods: The ethanol extract of Angelica gigas root was suspended in water and then partitioned with dichloromethane (MC Fr.), ethyl acetate (EA Fr.) and butanol (BuOH Fr.), sequentially. The antioxidant activities of solvent fractions of Angelica gigas root were evaluated for radical scavenging activity against stable free radicals (2,2-diphenyl-1-picrylhydrazyl) DPPH, hydrogen peroxide and superoxide anion radicals. In addition the antioxidant activities of solvent fractions of Angelica gigas root against peroxyl radicals, hydroxyl radicals and peroxynitrites were determined by the total oxy-radical scavenging capacity (TOSC) assay. Results: Among the solvent fractions of MC Fr., EA Fr., and BuOH Fr., BuOH Fr. was found to have stronger antioxidant activity with $IC_{50}$ values of 59.72, 14.36, 30.96 and $44.75\;{\mu}g/m{\ell}$ on the DPPH radical, nitrite, superoxide anion and hydrogen peroxide, respectively, than BHA used as a positive control. Moreover, specific TOSC values(564.8, 276.4 and 405.5 TOSC/mM) of BuOH fr. against peroxyl radicals, hydroxyl radical and peroxynitrite were 4 times higher than GSH (136.5, 67.4 102.6 TOSC/mM) used as a positive control. Conclusions: These results suggest that the BuOH fr. of Angelica gigas root has a high antioxidant activity and can be useful to develop functional food against oxidative stress conditions.

• 대한화장품학회지
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• 제48권3호
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• pp.265-273
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• 2022

본 연구에서는 한국산 겨우살이 (Viscum album L. var. coloratum)이 아토피 피부염과 관련된 염증성 사이토카인에 영향을 미치는지 여부를 알아보았다. 실험에는 헥산, 부탄올, 에틸아세테이트, 메틸렌클로라이드, 총 4 가지 분획물을 사용하였으며, RAW264.7 마우스 대식세포와 RBL-2H3 렛트 호중구를 이용하여 염증성 마커를 연구하였다. 실험 결과 에틸아세테이트 분획이 tumor necrosis factor alpha (TNF-α), interleukin(IL)-6, IL-4의 mRNA 발현 및 단백질 분비량을 감소시켰으나, 헥산 분획은 뚜렷한 효능이 없었다. 또한 부탄올 분획은 IL-4, IL-6의 mRNA 발현을 감소시켰고, 메틸렌클로라이드 분획은 IL-4와 TNF-α의 mRNA 발현을 감소시켰다. 결과적으로 한국산 겨우살이(V. album var. coloratum)가 아토피 피부염과 관련된 사이토카인 분비를 억제시켜 항염증 효과를 나타낼 수 있으므로, 이와 관련된 기능성 화장품 개발이 가능할 것으로 사료된다.

• 대한본초학회지
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• 제38권3호
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• pp.19-26
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• 2023

Objectives : Agrimoniae Herba is a herbal medicine widely distributed in Asia and contains flavonoids including catechin, quercitrin, rutin, hyperoside, and quercetin. This study aimed to investigate the anti-oxidant activity and skin barrier function of different solvent fractions (Hexane; methylene chloride, MC; ethyl acetate, EA; n-butanol, Bu; Water) obtained from Agrimoniae Herba. Methods : Anti-oxidant activity of different solvent fractions obtained from Agrimoniae Herba was investigated through total polyphenol contents, total flavonoid contents, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity measurements. Then, filament aggregating protein (Filaggrin), Type I collagen, ceramide synthase (CERS) 3, and CERS4 were analyzed to evaluate the skin barrier strengthening effect of different solvent fractions obtained from Agrimoniae Herba on UVB-stimulated HaCaT cells. Results : As a result of measuring total polyphenol contents, total flavonoid contents, DPPH free radical scavenging activity, and ABTS radical scavenging activity, antioxidant activity was found to be excellent in the order of EA > Bu > MC > Hexane > Water. As a result of measuring mRNA gene expression of Type I collagen, Filaggrin, CERS3, and CERS4 after UVB-stimulated was applied to HaCaT cells treated with different solvent fractions obtained from Agrimoniae Herba, it was found to increase significantly in the Bu-treated group. Conclusion : Our findings show that the Bu sample obtained from Agrimoniae Herba has excellent anti-oxidant ability, which increases Type I collagen, Filaggrin, and ceramide synthetase in UVB-stimulated HaCaT cells to control the skin barrier improvement effect.

• 대한의생명과학회지
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• 제12권4호
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• pp.443-450
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• 2006

To demonstrate their possibilities as an enhanced vaccine delivery system, protein-loaded Poly lactide glycolide copolymer (PLGA) microspheres were prepared with different physical characteristics. Ethyl acetate (EA) solvent extraction process was employed to prepare microspheres and the effects of process parameters on drug release properties were evaluated. The biodeuadability of microspheres was also evaluated by the pH change and GPC (Gel permeation chromatography). Primary IgG antibody responses in BALB/c mice were compared with protein saline solutions as negative controls and adsorbed alum suspensions as positive controls after single subcutaneous injection for in vivo studies. The microspheres showed a erosion with a highly porous structure and did not keep their spherical shape at 45 days and this result could be confirmed by GPC. In vitro release of proteinous drug showed initial burst effect in all batches of microspheres, followed by gradual release over the next 4 weeks. PLGA microspheres were degraded until 45 days and the secondary structure of OVA was not affected by the preparation method. Enzyme-linked immunosorbent assays demonstrated that the single subcutaneous administrations of OVA-loaded PLGA microspheres induced enhanced serum IgG antibody response in comparison to negative and positive controls. These results demonstrated that microspheres providing the controlled release of antigens might be useful in advanced vaccine formulations for the parenteral carrier system.

• 대한의생명과학회지
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• 제12권3호
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• pp.177-183
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• 2006

To demonstrate the effect of formulation conditions and evaluations of structural integrity from ovalbumin containing poly lactide glycolide copolymer (PLGA) microspheres for Vaccine delivery, OVA microspheres were prepared by a W/O/W multiple emulsion solvent extraction technique. Dichloromethan (DCM) and Ethyl acetate (EA) were applied as an organic phase and poly vinyl alcohol (PVA) as a secondary emulsion stabilizer. Microspheres were characterized for particle size, morphology (optical microscopy and Scanning Electron Microscope (SEM)). Protein denaturation was evaluated by size exclusion chromatography (SEC), SDS-PAGE and isoelectric focusing (IEF). Residual organic solvent was estimated by gas chromatography (GC) and differential scanning calorimetry (DSC). Optical photomicrograph and SEM revealed that micro spheres were typically spherical but various morphologies were observed. Mean particle size $(d_{vs})$ of microspheres were in the range of $3{\sim}50{\mu}m$. Also, The protein stability was not affected by the fonnulation process and residual organic solvent was beyond the detection below 0.1ppm. These results demonstrated that micro spheres might be a good candidate for the parenteral vaccine delivery system.

• 한국식품영양과학회지
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• 제43권5호
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• pp.705-711
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• 2014

피부미백 소재를 개발하기 위해 멜라닌 함량, 세포내 tyrosinase 활성의 측정 및 Western blotting 실험이 수행되었다. 백합(Lilium Oriental Hybrid 'Siberia') 뿌리의 80% 에탄올 추출물로부터 얻은 에틸아세테이트 분획물(R-EA)은 ${\alpha}$-melanocyte stimulating hormone(${\alpha}$-MSH)에 의해 멜라닌 생성이 유도된 B16/F10 흑색종 세포에서 농도 의존적으로 멜라닌 생성을 저해하였다. 정확하게 세포내 tyrosinase 활성과 멜라닌 함량은 에틸아세데이트 분획물 $100{\mu}g/mL$ 처리 시 ${\alpha}$-MSH 단독 처리군에 비해 각각 45%와 74%의 저해율을 보였다. ${\alpha}$-MSH에 의해 멜라닌 생성이 유도된 B16/F10 흑색종 세포에서 단백질 발현양상을 살펴본 결과 TRP-1이 가장 많이 억제된 양상을 확인할 수 있었고 이 결과는 세포내 tyrosinase 활성저해보다 멜라닌 생성저해가 더 많이 일어난 것과 일맥상통하는 것이다. 이를 종합해 볼 때 p-coumaric acid와 resveratrol 함량이 백합뿌리의 에탄올 추출물에 비해 많이 함유된 에틸아세테이트 분획물은 멜라닌 생성 유도물질에 의해 촉진된 ERK의 활성화를 억제하는 피부미백 소재로서 그 가치가 입증된다고 사료된다.

• 대한수의학회지
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• 제34권2호
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• pp.307-313
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• 1994

We have previously shown that crude water extract of Euonymus alatus (EA) had strong prophylactic effect against chemically induced-and tumor cell implanted-cancer, and that the mechanisms responsible for its antitumor effects were due to nonspecific enhancement of the NK cell activities and the cell mediated immunity. However, it was unknown that any components of crude extract did work so, since it consisted of several components. In this paper, we fractionated the crude watar EA-extract into several fraction such as hexane-, ethylether-, ethyl acetate-, n-butanol- and water soluble-fraction, and screened the immune regulating activities of each fraction by the evaluation of lymphokine production and activated lymphocyte proliferation. As a result of the component fraction of EA-extract, it was found that n-butanol fraction was a potent immunostimulator, and the remained water soluble fraction also contained some stimulator, But, other fraction did not showed any remarkable effect. It is therefore suggested that EA-glycosides in n-butanol fraction may be new one of the potent biological response modifiers. The present study was also undertaken in an efforts to investigate the effects of elm-bark(EB, Ulmus clavidiana var japonica), which has been used for curing ulcer and inflammation as a folk medicine without any kind of experimental evidence to support this, on the cellular- and humoral-immune responses, lymphocyte function and NK cell activities in mice. Regardless of time and duration of EB-treatment, Arthus reaction and antibody response to SRBC were not modified by EB, but delayed hypersensitivity to SRBC was significantly enhanced only when EB was treated prior to SRBC-sensitization. EB slightly inhibited the proliferation responses of splenocytes to PHA-stimulation, but it significantly augmented the responses of these cells to S aureus Cowan 1 and Con A-activation, and these effects were manifested only when EB was added at culture initiation. EB did not influence Ig secretion of spleen cells but it significantly augmented the Con A-induced 1L 2 and MIF production of splenocytes. NK cell activities of splenocytes were markedly riled when effector cells were pretreated with EB and this augmentation was dine to the increase of binding affinity of effector cells to target cells and the target cell lytic activities of effector cells. These results led to the conclusion that EB triggers increase of cellular immune responses, such as delayed hypersensitivitiy, lymphokine production and NK cell activities. Also these results suggested that EB contains potent immune stimulants, which may provide the rational basis for their therapeutic use as one of the new biological response modifiers.

• 생명과학회지
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• 제25권7호
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• pp.757-764
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• 2015

복분자는 당도가 낮고 신맛이 강해 과실주로 주로 제조되고 있으며, 복분자주 제조 후 부생되는 주박은 대부분 폐기되고 있다. 본 연구에서는 현재까지 생리활성이 보고된 바 없는 복분자주 주박을 대상으로 열수 추출물, ethanol 추출물 및 이들의 다양한 분획물을 조제하여 항균, 항산화 및 항혈전 활성을 평가하였다. 그 결과, 복분자주 주박의 ethylacetate (EA) 분획은 413~459 mg/g의 높은 polyphenol을 함유하고 있으며, 그람양성세균에 대한 강력한 성장억제 활성과, DPPH 음이온, ABTS 양이온에 대한 우수한 radical 소거능, nitrite 소거능 및 환원력을 나타내었다. 또한 혈액내의 thrombin, prothrombin, coagulation factor 에 대한 저해를 통한 혈액응고저해 활성이 우수하였다. 복분자주 주박의 butanol 분획은 우수한 항산화력과 함께 내인성 혈전 생성에 관련된 aPTT 연장활성을 나타내었으며, aspirin에 필적하는 강력한 혈소판 응집저해 활성을 나타내었다. 한편 복분자주 주박의 열수 추출물의 순차적 유기용매 분획 후의 물 잔류물은 aspirin보다 강력한 혈소판 응집저해를 나타내어 새로운 항혈소판제 개발 소재로 이용 가능함을 확인하였다. 본 연구결과는 폐기되고 있는 복분자주 주박을 이용한 고부가가치 식품 및 향장소재 개발이 가능함을 제시하고 있다.