• Title/Summary/Keyword: IC-MS/MS

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Pyrophen Produced by Endophytic Fungi Aspergillus sp Isolated from Piper crocatum Ruiz & Pav Exhibits Cytotoxic Activity and Induces S Phase Arrest in T47D Breast Cancer Cells

  • Astuti, Puji;Erden, Willy;Wahyono, Wahyono;Wahyuono, Subagus;Hertiani, Triana
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.615-618
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    • 2016
  • Ethyl acetate extracts obtained from culture of endophytic fungi Aspergillus sp isolated from Piper crocatum Ruiz & Pav, have been shown to possess cytotoxic activity against T47D breast cancer cells. Investigations were here conducted to determine bioactive compounds responsible for the activity. Bioassay guided fractionation was employed to obtain active compounds. Structure elucidation was performed based on analysis of LC-MS, $^1H$-NMR, $^{13}C$-NMR, COSY, DEPT, HMQC, HMBC data. Cytotoxity assays were conducted in 96 well plates against T47D and Vero cell lines. Bioassay guided isolation and chemical investigation led to the isolation of pyrophen, a 4-methoxy-6-(1'-acetamido-2'-phenylethyl)-2H-pyran-2-one. Further analysis of its activity against T47D and Vero cells showed an ability to inhibit the growth of T47D cells with IC50 values of $9.2{\mu}g/mL$ but less cytotoxicity to Vero cells with an $IC_{50}$ of $109{\mu}g/mL$. This compound at a concentration of 400 ng/mL induced S-phase arrest in T47D cells.

Anticoagulation and Anticancer Constituents from Eugenia caryophyllata Thunb

  • Han, Kyung-Min;Kim, Dong-Hyun;Ahn, Eun-Mi;Lee, Youn-Hyung;Chung, In-Sik;Kim, Dae-Keun;Kwon, Byoung-Mog;Kim, Sung-Hoon;Baek, Nam-In
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.82-88
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    • 2007
  • From the EtOAc fraction of Eugenia caryophyllata, four compounds were isolated through activity-guided silica gel column chromatography, From the result of spectroscopic data including NMR, MS and IR, the chemical structures of the compounds were determined as 1-allyl-4-hydroxy-3-methoxybezene acetate (eugenol acetate, 1), 1-allyl-4-hydroxy-3-methoxybezene (eugenol, 2), $3{\beta}-hydroxyolean-12-en-28-oic$ acid (oleanolic acid, 3) and $2{\alpha}$, $3{\beta}-dihydroxyolean-12-en-28-oic$ acid (maslinic acid, 4). Compounds 3 and 4 were isolated for the first time from this plant. Also, compounds 1, 2 and 3 exhibited relatively high platelet aggregation inhibitory activity with the $IC_{50}$ values of 0.24, 0.09 and 0.07 mM, respectively. Compound 2 significantly prolonged activated partial thromboplastin time (aPTT) with the value of $124{\pm}11.2$ seconds as compared to the control with the value of $37.5{\pm}2.2$ seconds at the concentration of 50 ${\mu}g/ml$. Compounds 1 and 3 revealed inhibitory activity on farnesyl protein transferase (FPTase) with the $IC_{50}$ values of 0.49 and 0.24 mM and compounds 1 and 2 highly inhibited the growth of rat-H-ras cells with the $Gl_{50}$ values of 6.63 and 5.70 ${\mu}M$, respectively.

Isolation of Angiotensin Converting Enzyme Inhibitor from Doenjang (전통된장으로부터 Angiotensin Converting Enzyme 저해물질의 분리)

  • Kim, Seung-Ho;Lee, Yun-Jin;Kwon, Dae-Young
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.848-854
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    • 1999
  • Inhibitory compounds of angiotensin converting enzyme (ACE) were separated from Doenjang (traditional Korean fermented soybean paste). Water extracts from Doenjang which showed ACE inhibitory activity were separated with gel permeation chromatography (GPC), in which two fractions with high ACE inhibitory activities were obtained. The first fraction from GPC was further isolated by semi-preparative reverse phase preparative-HPLC (high performance liquid chromatography) and 2-dimensional electrophoresis/thin layer chromatography (TLC). The purified spot had molecular weight of 759 daltons and ninhydrin-positive non-peptide. The second fraction from GPC was also further isolated by semi-preparative reverse phase HPLC and $NH_2-column$ HPLC. One fraction with high ACE inhibitory activity was purified and characterized. Molecular weight of this fraction by LC-MS was 272.34 daltons. The active fraction was identified as Arg-Pro with ACE $IC_{50}$ of $92\;{\mu}M$.

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Cytotoxic Triterpenoid from Rubus coreanus Miq

  • Lee, Dae-Young;Kim, Dong-Hyun;Bang, Myun-Ho;Song, Myoung-Chong;Kwak, Ho-Young;Yoo, Ki-Hyun;Chung, In-Sik;Kim, Kyong-Tai;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.275-280
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    • 2007
  • Dried unripe fruits of Rubus coreanus Miq. were extracted with 80% aqueous MeOH and the concentrated extract was partitioned with EtOAc and $H_2O$. From the EtOAc fraction, four triterpenoids were isolated through repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the result of physico-chemical data including NMR, MS aud IR, the chemical structures of the compounds were determined as tormentic acid (1), myrianthic acid (2), hovenic acid (3) and 2${\alpha}$,3${\beta}$,19${\beta}$,23-tetrahydroxylolean-12-en-28-oic acid (4). Compounds 3 and 4 were isolated for the first time from this plant. All isolated compounds were evaluated for cytotoxic activity against human colon carcinoma cells using in vitro three-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, compound 3 showed a higher cytotoxicity ($IC_{50}$ = 7.8 ${\mu}M$) than doxorubicin ($IC_{50}$= 50 ${\mu}M$).

Isolation of Biologically Active Compounds from the Flower Petals of Carthamus tinctorius L. (홍화(Carthamus tinctorius L.)로부터 활성물질의 분리)

  • Kim, Yung-Hee;Ahn, Eun-Mi;Bang, Myun-Ho;Nam, Ji-Youn;Kwon, Byung-Mok;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.41 no.2
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    • pp.197-200
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    • 1998
  • The MeOH extracts obtained from the flower petals of Carthamus tinctorius were solvent-fractionated with EtOAc, n-BuOH, and $H_2O$, successively. From the n-BuOH extract 2 flavonoid compounds were isolated through the repeated silica gel column chromatographies. From not only the results of physico-chemical data including HMBC but also the adaptation of acid hydrolysis, the chemical structures of the compounds were determined as $3-O-[{\beta}-D-glucopyranosyl(1{\rightarrow}2)\;{\beta}-D-glucopyranosyl]\;kaempferol$ and $3-O-[{\alpha}-L-rhamnopyranosyl(1{\rightarrow}6)\;{\beta}-D-glucopyranosyl]\;kaempferol$. The compounds exhibited $IC_{50}$ values in Grab2-Shc activity to be 43 and $47{\mu}g/ml$, respectively.

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Purification of Anti-obesity Lipase Inhibitor from the Fruiting Body of Phellinus linteus (상황버섯 자실체로부터 항비만성 라이페이스 저해물질의 정제)

  • Lee, Jong-Kuk;Song, Jung-Hwa;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.38 no.1
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    • pp.57-61
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    • 2010
  • For development of anti-obesity nutraceuticals from mushrooms, new anti-obesity lipase inhibitor from Phellinus linteus was purified by systematic solvent extraction, TLC and HPLC and characterized. Methanol extract from P. linteus most effectively inhibited(72.5%) porcine pancreatic lipase and ethyl acetate fraction showed the highest inhibitory activity in the systematic solvent extraction. A lipase inhibitor from the ethyl acetate fraction was purified through following steps including 3 times silica gel chromatography and preparative HPLC. The purified lipase inhibitor was a yellowish geen powder and its $IC_{50}$ value was 175 ng. Its molecular weight by MALDI-TOF-MS was 523.06 Da and showed maximal absorption spectrum at 225.1 nm.

Nitric Oxide Production Inhibitory Effects of Three Caffeoylquinic Acids Isolated from Hot Water Extract of Eriobotrya japonica L. Leaves (비파엽 열수 추출물에서 분리한 Caffeoylquinic Acid 3 종의 Nitric Oxide 생성 억제 효과)

  • Kim, Sun Min;Kim, A Young;Lee, Kyoung In
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.4
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    • pp.245-253
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    • 2020
  • Background: Research on hot water extracts of medicinal plants that are easily applicable in the clinical setting is essential. To confirm the anti-inflammatory-related active compounds present in the hot water extract of Eriobotrya japonica leaves, ability to inhibit nitric oxide (NO) production was measured and active compounds isolated from the extract were analyzed. Methods and Results: Sovent fractionation by solvent was performed to identify the active compounds present in the hot water extract, and the ability of the extract and the fractions obtained to inhibit NO production was measured. Subsequently, based on the results of liquid chromatography (LC) profile analysis of the n-butanol fraction that had a relatively high inhibitory ability of NO production, six subfractions were separated around the main peak. Among the separated subfractions spectra from mass spectroscopy (MS) were analyzed and standard comparisons were performed on the compounds of the three main peaks on the chromatogram. NO production inhibitory activity of subfraction 2 identified as neochlorogenic acid was the highest with an IC50 of 18.49 ㎍/㎖ followed by that of subfraction 5 identified as cryptochlorogenic acid with IC50 of 25.82 ㎍/㎖. Conclusions: Our result, it was confirmed that several caffeoylquinic acids, including neochlorogenic acid and cryptochlorogenic acid present in the hot water extract of E. japonica leaves have an important role as compounds exhibiting anti-inflammatory activity.

Antimalarial Activity and Phytochemical Profile of Ethanolic and Aqueous Extracts of Bidara Laut (Strychnos ligustrina Blum) Wood

  • MANURUNG, Harisyah;SARI, Rita Kartika;SYAFII, Wasrin;CAHYANINGSIH, Umi;EKASARI, Wiwied
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.5
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    • pp.587-596
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    • 2019
  • This study aimed to determine the antimalarial effect of the Strychnos ligustrina (SLW) wood extracts and to analyze its phytochemicals. The SLW powder samples were macerated with 100% ethanol (E100), 75% ethanol (E75), 50% ethanol (E50), 25% ethanol (E25), and aqueous (A100). The extracts were analyzed by LCMS/MS, and its in-vitro antimalarial activity was tested with Plasmodium falciparum. The results showed that the extract yields of E100, E75, E50, E25, and A100 were 4.3, 5.2, 5.3, 4.7, and 3.6%, respectively. The antimalarial activities of the A100, E25, E50, and E75 extracts were classified as active with $IC_{50}$ values of 38.6, 42.6, 42.9, and $43.7{\mu}g/mL$, respectively. But, the antimalarial activity of the E100 extract was classified as slightly active with $IC_{50}$ values of $87.4{\mu}g/mL$. The dominant compounds contained in the extracts of A100, E25, E50, E75, and E100 was the alkaloid compound, namely brucine with relative concentrations of 24.96, 24.55, 21.33, 11.79, and 11.62%, respectively.

Chemical composition, antioxidant and antifungal activities of rhizome essential oil of Kaempferia parviflora Wall. ex Baker grown in Vietnam

  • Dang-Minh-Chanh Nguyen;Thi-Hoan Luong;Tien-Chung Nghiem;Woo-Jin Jung
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.15-22
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    • 2023
  • The aim of the present study was to evaluate the chemical composition and antioxidative activity of rhizome essential oil of Kaempferia parviflora Wall. ex Baker. The essential oil extracted by hydrodistillation was chemically profiled by GC/MS analysis. The antioxidative activity was determined and evaluated spectroscopically by the 2,2-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging assays. According to the results, the major essential oil components were camphene (18.03%), β-pinene (14.25%), a-pinene (12.38%), endo-borneol (10.23%), β-copaene (8.38%), and linalool (8.20%). K. parviflora rhizome oil possessed antioxidant potential, exhibiting DPPH and ABTS radical scavenging activities as high as 80.90 and 94.04%, respectively, at a concentration of 10 mg/mL. The corresponding IC50 values were 0.451±0.051 and 0.527±0.022 mg/mL, respectively (IC50 values for ascorbic acid, as the standard, were 0.209±0.016 and 0.245±0.022 mg/mL, respectively). The mycelium of F. oxysporum was distorted and collapsed when treated with 0.5 mg/mL of the EO of K. parviflora rhizome for 3 days treatment, which may provide an important information for exploring the metabolism of the fungicide K. parviflora rhizome and its derived compounds against F. oxysporum. This study provides the chemical properties of the essential oil of K. parviflora rhizome grown in Vietnam and their potential antioxidant and antifungal activities.

HPLC, NMR Based Characterization, Antioxidant and Anticancer Activities of Chemical Constituents from Therapeutically Active Fungal Endophytes

  • Waqas Hussain Shah;Wajiha Khan;Sobia Nisa;Michael H.J. Barfuss;Johann Schinnerl;Markus Bacher;Karin Valant-Vetschera;Ashraf Ali;Hiba-Allah Nafidi;Yousef A. Bin Jardan;John P. Giesy
    • Journal of Microbiology and Biotechnology
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    • v.34 no.7
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    • pp.1452-1463
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    • 2024
  • Fungi generate different metabolites some of which are intrinsically bioactive and could therefore serve as templates for drug development. In the current study, six endophytic fungi namely Aspergillus flavus, Aspergillus tubigenesis, Aspergillus oryzae, Penicillium oxalicum, Aspergillus niger, and Aspergillus brasiliensis were isolated and identified from the medicinal plant, Silybum marianum. These endophytic fungi were identified through intra transcribed sequence (ITS) gene sequencing. The bioactive potentials of fungal extracts were investigated using several bioassays such as antibacterial activity by well-diffusion, MIC, MBC, anti-biofilm, antioxidant, and haemolysis. The Pseudomonas aeruginosa PAO1 was used to determine the antibiofilm activity. The ethyl acetate extract of Aspergillus flavus showed strong to moderate efficacy against Staphylococcus aureus, Escherichia coli, P. aeruginosa, and Bacillus spizizenii. Aspergillus flavus and Aspergillus brasiliensis exhibited significant antibiofilm activity with IC50 at 4.02 and 3.63 mg/ml, while A. flavus exhibited maximum antioxidant activity of 50.8%. Based on HPLC, LC-MS, and NMR experiments kojic acid (1) and carbamic acid (methylene-4, 1-phenylene) bis-dimethyl ester (2) were identified from A. flavus. Kojic acid exhibited DPPH free radical scavenging activity with an IC50 value of 99.3 ㎍/ml and moderate activity against ovarian teratocarcinoma (CH1), colon carcinoma (SW480), and non-small cell lung cancer (A549) cell lines. These findings suggest that endophytic fungi are able to produce promising bioactive compounds which deserve further investigation.