• Title/Summary/Keyword: ethyl acetate extract.

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Isolation and Identification of Active Antimicrobial Substance against Listeria monocytogenes from Ruta graveolens Linne (운향으로부터 Listeria monocytogenes에 대한 항균 활성 물질의 분리 및 구조동정)

  • Ahn, Yong-Seon;Shin, Dong-Hwa;Baek, Nam-In
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1379-1388
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    • 2000
  • Ethanol extracts from Ruta graveolens Linne exhibited strong antimicrobial activities by disc diffusion method against 5 strains of Listeria monocytogenes (ATCC 19111, ATCC 19112, ATCC 19113, ATCC 19114 and ATCC 15313). Ethanol extract from Ruta graveolens Linne was subsequently fractionated by n-hexane, chloroform, ethyl acetate and water. Chloroform fraction of Ruta graveolens Linne showed strong growth inhibition at concentrations as low as 40 ppm level in broth culture medium against 5 strains of L. monocytogenes for 72 hr at $30^{\circ}C$. Single substance(RTG1-1) was isolated by silica gel column chromatography from chloroform fraction of Ruta graveolens Linne. RTG1-1 showed a strong bactericidal activity against L. monocytogenes at a concentration of 20 ppm level. Purified RTG1-1 was identified as gravacridonechlorine by analyses of EI-Mass, $^1H-NMR$ and $^{13}C-NMR$.

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A comparative study of Sargassum horneri Korea and China strains collected along the coast of Jeju Island South Korea: its components and bioactive properties

  • Kim, Hyun-Soo;Sanjeewa, K.K. Asanka;Fernando, I.P. Shanura;Ryu, BoMi;Yang, Hey-Won;Ahn, Ginnae;Kang, Min Cheol;Heo, Soo-Jin;Je, Jun-Geon;Jeon, You-Jin
    • ALGAE
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    • v.33 no.4
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    • pp.341-349
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    • 2018
  • Sargassum horneri is edible brown seaweed abundant along the coasts of Jeju Island, South Korea. In addition to the native S. horneri population, a large amount of S. horneri has been found to invade Jeju Island from the east coast of China. Thus, S. horneri of both Korea (SK) and China (SC) strains now inhabits along with the shore of Jeju Island and have become a threat to the coastal biodiversity. However, they could be used in obtaining functional ingredients for industrial level applications provided an optimized cost effective strategy. In the present study, we compared SK and SC strains for the extraction efficiency, components, antioxidant, and anti-inflammatory properties of 80% methanolic extracts and their partially purified fractions. According to the results, two strains indicated similar bioactive properties such as DPPH and alkyl radical scavenging activity as well as anti-inflammatory activities on lipopolysaccharide-stimulated RAW 264.7 cells. The yield of 80% methanol extract from SC was higher than SK. However, the yields of the ethyl acetate and chloroform fractions from SK were higher than those of SC strain. The major peaks in the high-performance liquid chromatography chromatograms, which was identified as Apo-9 fucoxanthinone, indicated that both methanolic extracts of SK and SC contains major target peaks but with different amounts. This study might be useful for developing functional materials from SC and SK in future.

Development and validation of a qualitative GC-MS method for THCCOOH in urine using injection-port derivatization

  • Sim, Yeong Eun;Kim, Ji Woo;Kim, Jin Young
    • Analytical Science and Technology
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    • v.34 no.2
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    • pp.68-77
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    • 2021
  • Cannabis is one of the most abused drugs in Korea. The main psychoactive component in cannabis, Δ9-tetrahydrocannabinol, is metabolized to 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH) and THCCOOH-glucuronide (THCCOOH-glu) in the human liver, whereby the amount of THCCOOH-glu found in urine is twice as high as that of THCCOOH. The analytical process adapted by the majority of urine drug-testing programs involves a two-step method consisting of an initial immunoassay-based screening test followed by a confirmatory test if the screening test result is positive. In this study, a qualitative gas chromatography-mass spectrometry (GC-MS) method was developed and validated for the detection of THCCOOH in human urine, where THCCOOH-glu was converted into THCCOOH by alkaline hydrolysis. For purification of the urine extract prior to instrumental analysis, high-speed centrifugation was used to minimize interference. In addition, an injection-port derivatization method using ethyl acetate and N,O-bis(trimethylsilyl)-trifluoroacetamide containing 1 % trimethylchlorosilane was employed to reduce the time required for derivatization, and an aliquot of the final solution was injected into the GC-MS. The method was validated by measuring the selectivity, limit of detection (LOD), and repeatability. The sensitivity, specificity, precision, accuracy, Kappa, F-measure, false positive, and false negative rate were determined by comparing the GC-MS results with those obtained using the immunoassay. The LOD was determined to be 0.32 ng/mL, while the repeatability was within 9.1 % for THCCOOH. Furthermore, a comparison study was carried out, whereby the screening immunoassay exhibited a sensitivity of 86.4 % and a specificity of 100 % compared to GC-MS. The applicability of the developed method was examined by analyzing spiked urine and forensic urine samples obtained from suspected cannabis abusers (n = 221).

Amelioration of experimental autoimmune encephalomyelitis by Ishige okamurae

  • Ahn, Meejung;Kim, Jeongtae;Yang, Wonjun;Choi, Yuna;Ekanayake, Poornima;Ko, Hyunju;Jee, Youngheun;Shin, Taekyun
    • Anatomy and Cell Biology
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    • v.51 no.4
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    • pp.292-298
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    • 2018
  • Experimental autoimmune encephalomyelitis (EAE) is a T-cell-mediated autoimmune central nervous system disease characterized by inflammation with oxidative stress. The aim of this study was to evaluate an anti-inflammatory effect of Ishige okamurae on EAE-induced paralysis in rats. An ethanolic extract of I. okamurae significantly delayed the first onset and reduced the duration and severity of hind-limb paralysis. The neuropathological and immunohistochemical findings in the spinal cord were in agreement with these clinical results. T-cell proliferation assay revealed that the ethyl-acetate fraction of I. okamurae suppressed the proliferation of myelin basic protein reactive T cells from EAE affected rats. Flow cytometric analysis showed $TCR{\alpha}{\beta}^+$ T cells was significantly reduced in the spleen of EAE rats with I. okamurae treatment with concurrent decrease of inflammatory mediators including tumor necrosis $factor-{\alpha}$ and cyclooxygenase-2. Collectively, it is postulated that I. okamurae ameliorates EAE paralysis with suppression of T-cell proliferation as well as decrease of pro-inflammatory mediators as far as rat EAE is concerned.

Antioxidant and anti-inflammatory activity of parts of Rhus javanica L. (붉나무의 부위 별 항산화 및 항염증 활성)

  • Choi, Ji-Soo;Han, Sang-Don;Jang, Tae-Won;Lee, Seung-Hyun;Park, Jae-Ho
    • Journal of Applied Biological Chemistry
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    • v.62 no.2
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    • pp.195-202
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    • 2019
  • Rhus javanica L. is Anacardiaceae plant distributed in East Asia. We evaluated the antioxidant activity and antiinflammatory effect of leaf, branch, root of ethyl acetate fraction from R. javanica. To confirm effective each extraction, The antioxidant activity was evaluated using 1,1-Diphenyl-2-picryl-hydrazyl and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) scavenging activity assays, and the anti-inflammatory activity was evaluated based on inhibitory activities on the protein and mRNA expression of iNOS and COX-2 in LPS-induced RAW264.7 cells. The phenolic compounds content of each extract was analyzed with Folin reagents and HPLC/PDA method. The gallic acids were identified and quantified. The roots of R. javanica showed strong antioxidant activity. Its total phenolic compounds content were higher than the orders. In addition, anti-inflammatory activity inhibited the protein and mRNA expression of nitric oxide production factor, following the same pattern as contents of phenolic compounds included gallic acid and its antioxidant activity. In conclusion, R. javanica showed effective antioxidant and anti-inflammatory activity. Especially, the roots were evaluated to be highly valuable as a natural resource for reducing inflammation.

Fermentation enhances the antioxidant and anti-inflammatory effects of Bat Faeces (Ye Ming Sha) via the ERK, p38 MAPK and NF-κB signaling pathways in RAW 264.7 cells

  • Lee, Han-Saem;Chon, So-Hyun;Kim, Min-A;Park, Jeong-Eun;Lim, Yu-Mi;Kim, Eun-Jeong;Son, Eun-Kyung;Kim, Sang-Jun;So, Jai-Hyun
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.57-66
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    • 2019
  • The ethyl acetate fraction of Bat Faeces (Ye Ming Sha: natural products used in Chinese Medicine) after fermentation (EFBF-AF) showed enhanced anti-oxidative effects in 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt assays. Fermentation of the Bat Faeces by using the crude enzyme extract from Aspergillus kawachii, significantly increased the anti-inflammatory effects. Fermented Bat Faeces markedly inhibited nitric oxide production, inducible nitric oxide synthase, and cyclooxygenase-2 expression in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. The EFBF-AF reduced the nuclear translocation of nuclear factor kappa B ($NF-{\kappa}B$) via $IKK{\alpha}$ and $I{\kappa}B{\alpha}$ phosphorylation, and decreased the phosphorylated the extracellular signal-regulated kinases (ERK) and p38 expression in LPS-treated RAW 264.7 macrophages. In addition, the EFBF-AF suppressed the expression of pro-inflammatory genes, such as interleukin-$1{\beta}$, interleukin-6, and tumor necrosis $factor-{\alpha}$. These results suggest that fermented Bat Faeces may suppress pro-inflammatory responses in LPS-stimulated RAW 264.7 macrophages cells via ERK, p38 mitogen-activated protein kinase and $NF-{\kappa}B$ signaling pathways.

Genome Analysis and Optimization of Caproic Acid Production of Clostridium butyricum GD1-1 Isolated from the Pit Mud of Nongxiangxing Baijiu

  • Min Li;Tao Li;Jia Zheng;Zongwei Qiao;Kaizheng Zhang;Huibo Luo;Wei Zou
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1337-1350
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    • 2023
  • Caproic acid is a precursor substance for the synthesis of ethyl caproate, the main flavor substance of nongxiangxing baijiu liquor. In this study, Clostridium butyricum GD1-1, a strain with high caproic acid concentration (3.86 g/l), was isolated from the storage pit mud of nongxiangxing baijiu for sequencing and analysis. The strain's genome was 3,840,048 bp in length with 4,050 open reading frames. In addition, virulence factor annotation analysis showed C. butyricum GD1-1 to be safe at the genetic level. However, the annotation results using the Kyoto Encyclopedia of Genes and Genomes Automatic Annotation Server predicted a deficiency in the strain's synthesis of alanine, methionine, and biotin. These results were confirmed by essential nutrient factor validation experiments. Furthermore, the optimized medium conditions for caproic acid concentration by strain GD1-1 were (g/l): glucose 30, NaCl 5, yeast extract 10, peptone 10, beef paste 10, sodium acetate 11, L-cysteine 0.6, biotin 0.004, starch 2, and 2.0% ethanol. The optimized fermentation conditions for caproic acid production by C. butyricum GD1-1 on a single-factor basis were: 5% inoculum volume, 35℃, pH 7, and 90% loading volume. Under optimal conditions, the caproic acid concentration of strain GD1-1 reached 5.42 g/l, which was 1.40 times higher than the initial concentration. C. butyricum GD1-1 could be further used in caproic acid production, NXXB pit mud strengthening and maintenance, and artificial pit mud preparation.

Steroid Components of Marine-Derived Fungal Strain Penicillium levitum N33.2 and Their Biological Activities

  • Chi K. Hoang;Cuong H. Le; Dat T. Nguyen;Hang T. N. Tran;Chinh V. Luu;Huong M. Le;Ha T. H. Tran
    • Mycobiology
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    • v.51 no.4
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    • pp.246-255
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    • 2023
  • Genus Penicillium comprising the most important and extensively studied fungi has been well-known as a rich source of secondary metabolites. Our study aimed to analyze and investigate biological activities, including in vitro anti-cancer, anti-inflammatory and anti-diabetic properties, of metabolites from a marine-derived fungus belonging to P. levitum. The chemical compounds in the culture broth of P. levitum strain N33.2 were extracted with ethyl acetate. Followingly, chemical analysis of the extract leaded to the isolation of three ergostane-type steroid components, namely cerevisterol (1), ergosterol peroxide (2), and (3β,5α,22E)-ergosta-6,8(14),22-triene-3,5-diol (3). Among these, (3) was the most potent cytotoxic against human cancer cell lines Hep-G2, A549 and MCF-7 with IC50 values of 2.89, 18.51, and 16.47 ㎍/mL, respectively, while the compound (1) showed no significant effect against tested cancer cells. Anti-inflammatory properties of purified compounds were evaluated based on NO-production in LPS-induced murine RAW264.7 macrophages. As a result, tested compounds performed diverse inhibitory effects on NO production by the macrophages, with the most significant inhibition rate of 81.37±1.35% at 25 ㎍/mL by the compound (2). Interestingly, compounds (2) and (3) exhibited inhibitory activities against pancreatic lipase and α-glucosidase enzymes in vitro assays. Our study brought out new data concerning the chemical properties and biological activities of isolated steroids from a P. levitum fungus.

Inhibition of Food-derived Lactic Acid Bacterial Biofilm Formation Using Eisenia bicyclis-derived Nanoparticles (식품 유래 Biofilm 형성 유산균에 대한 대황(Eisenia bicyclis) 유래 Nanoparticle의 Biofilm 형성 저해)

  • Do Kyung Oh;Fazlurrahman Khan;Seul-Ki Park;Du-Min Jo;Kyung-Jin Cho;Geum-Jae Jeong;Yeon-Ju Sim;Jeong Mi Choi;Jae-Ho Woon;Young-Mog Kim
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.57 no.2
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    • pp.129-136
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    • 2024
  • Lactic acid bacteria (LAB) growth in processed meat products produces slime. In this study, 10 different biofilm-forming LAB, including Leuconostoc mesenteroides, Lacticaseibacillus paracasei, Levilactobacillus brevis, Lactiplantibacillus plantarum, Leuconostoc citreum, Weissella viridescens, and Latilactobacillus sakei, were isolated from various meat products and identified based on 16S rRNA gene analysis. To inhibit biofilm formation by LABs, Eisenia bicycles methanolic extract (EB) and ethyl acetate soluble fraction (EA) were used as antibacterial and antibiofilm agents, respectively. Furthermore, EA and EB were employed to synthesize gold nanoparticles (AuNPs) such as EB-AuNPs and EA-AuNPs, which could serve as antibiofilm agents against the isolated LAB. These findings demonstrate that EA, EB-AuNPs, and EA-AuNPs exhibit significant antibacterial activity against the isolated LAB. Furthermore, EB-AuNPs reduced L. citreum biofilm production, whereas EA-AuNPs inhibited L. mesenteroides and L. brevis biofilm formation. The current results suggest that EB-AuNPs and EA-AuNPs can be used as nanomaterials to inhibit LAB that form biofilms on meat products.

Cytoprotective Effect of Cinnamomum japonicum Siebold Branch Extracts via Blocking Oxidative Stress in Hepatocytes (간세포에서 산화적 스트레스 억제를 통한 생달가지 추출물의 세포보호 효과)

  • Ji Hye Yang
    • Herbal Formula Science
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    • v.31 no.4
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    • pp.283-293
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    • 2023
  • Objectives : Native to East Asia, Japan, and Korea, Cinnamomum japonicum Siebold (CJ) is renowned for its aromatic leaves and bark. We previously assessed the antioxidant activity of fractionated CJ branches (CJB:70% EtOH extract), including hexane (CJB1), chloroform (CJB2), ethyl acetate (CJB3), butanol (CJB4), and water (CJB5). Our findings revealed that CJB3 exhibited the highest antioxidant activity. Here, we aimed to investigate whether CJB3 possesses cytoprotective effects and induces the activity of antioxidant enzymes in hepatocytes. Methods : As HepG2 cells were the first to exhibit the key characteristics of hepatocytes, we investigated the hepatoprotective effects of CJB3 on HepG2 cells. Results : Before conducting the cell experiment, we checked that CJB3, up to a concentration of 100 ㎍/mL, did not exhibit cytotoxicity toward HepG2 cells. ROS production increased because of t-BHP treatment decreased in a concentration-dependent manner upon CJB3 treatment. We confirmed that CJB3 inhibited t-BHP-induced cell death. CJB3 was found to reverse the expression of proteins associated with t-BHP-induced apoptosis. We also observed that CJB3 induced Nrf2 phosphorylation and the nuclear translocation of Nrf2. And, CJB3 treatment caused a time-dependent enhancement of GCL and NQO1 protein expression. We further confirmed that CJB3 increased the expression of Nrf2 target genes, and this effect was associated with the activation of JNK, p38, and AMPK. Conclusion : CJB3 prevents t-BHP-induced oxidative stress and apoptosis and enhances the expression of Nrf2 target genes via JNK, p38, and AMPK activation. These results suggest that CJB3 is a promising candidate for the treatment of liver diseases.