• Title/Summary/Keyword: Chromatogram

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Bioassay-coupled LC-QTOF MS/MS to Characterize Constituents Inhibiting Nitric Oxide Production of Thuja orientalis

  • Park, Dawon;Shin, Hyeji;Byun, Youngjoo;Lee, Ki Yong
    • Natural Product Sciences
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    • v.27 no.4
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    • pp.293-299
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    • 2021
  • The ethyl acetate fractions prepared from the leaves of Thuja orientalis significantly inhibited nitric oxide (NO) production in lipopolysaccharide-stimulated BV2 microglial cells. According to bioassay-coupled LC-QTOF MS/MS, the components near 22 and 25 mins in the mass chromatogram highly inhibited NO production and were expected to be labdane diterpenes, and the active components were characterized via further isolation. The results of the NO production inhibitory assay of the isolated compounds correlated well with the results of bioassay-coupled LC-QTOF MS/MS. Among the identified constituents, NO production inhibitory activities of 16-hydroxy-labda-8(17),13-diene-15,19-dioic acid butenolide (2) and 15-hydroxypinusolidic acid (3) were newly reported. Taken together, these results demonstrated that LC-QTOF MS/MS coupled with NO production inhibition assay was a powerful tool for accurately predicting new anti-inflammatory constituents in the extracts from natural products. Moreover, it provided a potential basis for broadening the application of bioassay-coupled LC-QTOF MS/MS in natural product research.

Pharmacognostical Studies on the Folk Medicine "SaCheolNaMu" (민간약 사철나무의 생약학적 연구)

  • Park, Woo Sung;Chung, Hye-Jin;Bae, Ji-Yeong;Park, Jong Hee;Ahn, Mi-Jeong
    • Korean Journal of Pharmacognosy
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    • v.44 no.4
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    • pp.320-325
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    • 2013
  • "SaCheolNaMu" has been used as a Korean folk medicine for the jaundice, lumbago and uterine diseases. Although a crude drug related to this folk medicine is sold in traditional herbal market, the botanical origin of this drug has not been pharmacognostically confirmed yet. In this study, the morphological and anatomical characteristics of the stem of Euonymus species growing in Korea, i.e. Euonymus japonica and E. fortunei var. radicans were studied to clarify the botanical origin of "SaCheolNaMu". As a result, it was found that these two species could be discriminated by the morphological criteria such as the thickness of cuticles, the number of collenchyma cell layers, and the frequency of druse and resinous substance. According to these criteria, it was elucidated that the commercial folk medicine "SaCheolNaMu" was the stem of E. japonica. Meanwhile, HPLC-DAD analysis on the 70% ethanolic extracts of two species showed significantly different HPLC profiles each other. The molecular ions of three characteristic peaks shown in the chromatogram of two species were identified by ESI-MS, and their structures were estimated to be flavonol glycosides.

Epimerization of L-Arabinose for Producing L-Ribose (L-리보스 생산을 위한 L-아라비노스의 에피머반응)

  • Jeon, Young Ju;Song, Sung Moon;Lee, Chang Soo;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.49 no.5
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    • pp.628-632
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    • 2011
  • L-ribose has recently attracted interest as a starting material for antiviral drug. It could be obtained from L-arabinose by epimerization reaction. Epimerization reaction was carried out with molybdenium oxide or molybdic acid catalyst and methanol/water solution. Reaction temperature, methanol percentage, and catalyst kind were selected to find an optimum reaction condition. Ion exhange chromatography was used for separating epimerization reaction mixture, and then HPLC chromatogram of L-ribose fraction obtained to calculate the yield of the reaction. Shodex ion exchange HPLC column(Model SC1011) and Phenomenex Luna $NH_2$ HPLC column were compared to employ a convenient HPLC analysis. It was found that the usage of 20% methanol, $60^{\circ}C$, and 40 g/L molybdic acid gives the best reaction condition with a yield of 21%.

Analysis of Essential oil, Quantification of Six Glycosides, and Nitric Oxide Synthase Inhibition Activity in Caryopteris incana

  • Nugroho, Agung;Lee, Sang Kook;Kim, Donghwa;Choi, Jae Sue;Park, Kyoung-Sik;Song, Byong-Min;Park, Hee-Juhn
    • Natural Product Sciences
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    • v.24 no.3
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    • pp.181-188
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    • 2018
  • Caryopteris incana (Verbenaceae) has been used to treat cough, arthritis, and eczema in Oriental medicine. The two fractions ($CHCl_3-$ and BuOH fractions) and the essential oil of the plant material were subjected to the inducible nitric oxide synthase (iNOS) assay. The $IC_{50}$ of the $CHCl_3$ fraction and the essential oil on LPS-induced macrophage RAW 264.7 cells were $16.4{\mu}g/mL$ and $23.08{\mu}g/mL$, respectively. On gas chromatography (GC)-mass spectroscopy (MS) analysis, twenty-five components representing 85.5% amount of total essential oil were identified. On the chromatogram, three main substances, trans-pinocarveol, cis-citral, and pinocarvone, occupied 18.8%, 13.5% and 18.37% of total peak area. Furthermore, by HPLC-UV analysis, six compounds including one iridoid (8-O-acetylharpagide)- and five phenylethanoid glycosides (caryopteroside, acteoside, phlinoside A, 6-O-caffeoylphlinoside, and leucosceptoside A) isolated from the BuOH fraction were quantified. The content of six compounds were shown as the following order: caryopteroside (162.35 mg/g) > 8-O-acetylharpagide (93.28 mg/g) > 6-O-caffeoylphlinoside (28.15 mg/g) > phlinoside (22.60 mg/g) > leucosceptoside A (16.87 mg) > acteoside (7.05 mg/g).

Identification of Natural dyes used in 16th pink Dallryeong (Official's robe in Joseon Dynasty) Excavated from Cheonan, Chungnam (충남 천안시 출토 16세기 분홍 단령에 사용된 염재 동정)

  • Chae, Jeongmin;Ryu, Hyo-Seon
    • Journal of Conservation Science
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    • v.31 no.3
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    • pp.299-308
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    • 2015
  • Aim of this study is to identify dyestuff of the Dallryeong(official's robe in Joseon Dynasty, 16th century) excavated from Yuryang-dong, Cheonan, in 1996. For this purpose, extracted dyestuffs from Dallryeong fabric and from natural dyestuffs for red color(safflower, Sapanwood, Madder) which are presumed to have been used in the Dallryeong, are analyzed and compared by high performance liquid chromatography(HPLC). As a result, HPLC chromatogram of extracts of the Dallryeong's dyestuff and safflower are showed a peak at 17.5 minutes. The UV/Vis spectra of the samples are showed the maximum absorption wavelength at 519nm. This result is identical with the analysis of the previous studies on red dyestuff of safflower. In addition, the analysis of Mass Spectrometry(MS) showed the identical result of the peak with m/z 910. Following these results, excavated pink Dallryeong were considered to have been dyed with safflower.

N-oleoyl-D-erythro-sphingosine-based Analysis of Ceramide by High Performance Liquid Chromatography and Its Application to Determination in Diverse Biological Samples

  • Lee, Youn-Sun;Choi, Heon-Kyo;Yoo, Jae-Myung;Choi, Kyong-Mi;Lee, Yong-Moon;Oh, Sei-Kwan;Kim, Tack-Joong;Yun, Yeo-Pyo;Hong, Jin-Tae;Okino, Nozomu;Ito, Makoto;Yoo, Hwan-Soo
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.273-281
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    • 2007
  • Ceramide is involved in cell death as a lipid mediator of stress responses. In this study, we developed an improved method of ceramide quantification based on added synthetic ceramide and thin layer chromatography (TLC) separation, and applied to biological samples. Lipids were extracted from samples spiked with N-oleoyl-D-erythro-sphingosine ($C_{17}$ ceramide) as an internal standard. Ceramide was resolved by TLC, complexed with fatty-acidfree bovine serum albumin (BSA), and deacylated by ceramidase (CDase). The released sphingosine was derivatized with o-phthalaldehyde (OPA) and measured by high performance liquid chromatography (HPLC). The limit of detection for ceramide was about 1-2 pmol and the lower limit of quantification was 5 pmol. Ceramide recovery was approximately 86-93%. Ceramide concentrations were determined in biological samples including cultured cells, mouse tissues, and mouse and human plasma. TLC separation of ceramide provides HPLC chromatogram with a clean background without any interfering peaks and the enhanced solubility of ceramide by BSAceramide complex leads to the increased deacylation of ceramide. The use of an internal standard for the determination of ceramide concentration in these samples provides an accurate and reproducible analytical method, and this method can be applicable to diverse biological samples.

Analysis of Organochlorine Pesticide, in the Presence of Polychlorinated Biphenyls(PCBs) I. Florisil Column Separation of the Pesticides-PCBs Mixture (Polychlorinated Biphenyls(PCBs) 존재하에 유기염소계 살충제의 잔류분 분석 1. Florisil Column에 의한 농약과 PCBs의 분리)

  • Park C. K.;Lee C. Y.;Park R. D.
    • Korean journal of applied entomology
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    • v.16 no.4 s.33
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    • pp.221-227
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    • 1977
  • Polychlorinated Biphenyls (PCBs) interfere with gas chromatographic analysis of multiple organochlorine pesticide residues. In the present work, existing Florisil column chromatographic method has been modified as to improve separation of organochlorine pesticides and their metabolites from PCBs. It was amply demonstrated that separation of $\alpha-BHC,\; \gamma-BHC$ Heptachlor epoxide Dieldrin, p.p-DDD, p.p'-DDT from PCBs such as Aroclor 1254 is complete and recovery of the pesticides is found quantitative. Aldrin and Heptachlor in the Aroclor eluant can be separately analyzed by comparison of the chromatographic pattern of standard Aroclor 1254 with that of Aroclor 1254 a dmixed with the two pesticides. The Florsil column technique can be utilized in the routine evaluation of the organochlorine pesticide residues by gas chromatography.

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Porduction and Enzymatic Characteristics of ${\alpha}-Galactosidase$ from Aspergillus niger (Aspergillus niger에 의한 ${\alpha}-Galactosidase$의 생산 및 효소적 특성)

  • Chun, Hyang-Sook;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.20 no.1
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    • pp.79-84
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    • 1988
  • ${\alpha}$-Galactosidase from Aspergillus niger as a possible enzyme for removal of flatulence factors in soybean foods was produced the highest in 120 hours in either Czapeck-Dox liquid medium or wheat bran solid medium. The most efficient carbon and nitrogen sources in Czapeck-Dox medium were raffinose and sodium nitrate, respectively, whereas the addition of the sources showed negative effects in wheat bran. pH optima for enzyme activity and stability were 4.0-5.0 and 3.5-6.5, respectively, and optimum temperature for stability was $40-50^{\circ}C$. Upon reaction on p-nitrophenyl-${\alpha}$-D-galactoside, Michaelis constant was 0.42 mM and maximum velocity was 152 ${\mu}moles$ substrate/minute/kg solid medium. Mercuric chloride acted as a strong noncompetitive inhibitor and p-chloromercuribenzoate, even in low concentration, acted as a competitive inhibitor. Crude ${\alpha}$-galactosidase hydrolyzed raffinose and stachyose completely, giving spots of monosaccharides only on thin-layer chromatogram.

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Studies on Naringinase of Mold - Part 2. Purification of Aspergillus Naringinase - (사상균 Naringin 분해효소에 관한 연구 - 제 2 보 Aspergillus 속 Naringin 분해효소의 정제에 관하여 -)

  • Ki, Woo-Kyung;Kim, Jong-Kyu;Kim, Myung-Chan
    • Korean Journal of Food Science and Technology
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    • v.5 no.2
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    • pp.78-83
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    • 1973
  • The naringin hydrolyzing enzyme has been purified from the culture filtrate of the mold Aspergillus S-1 which selected to remove the bitter test of the orange or citrus fruits industrily. In a view of purity naringinase was more effectively purified in order of molecular sieving on Sephadeex G-200, starach gel electrophoresis, chromatography or a DEAE-Cellulose column and fractional precipitation by ammonium sulfate. The purified enzyme is homogeneous in paper electrophoresis from a culture filtrate by treatment fractional precipitation with ammonium sulfate, DEAE-Cellulose treatment and Sephadex-200 column chromatography and it hydrolyse only naringin to purunin.

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Amino Acids Analysis of Silk Fibroins among Varieties (가잠견사 Fibroin의 품종별 Amino산 분석)

  • 최병희;김한수
    • Journal of Sericultural and Entomological Science
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    • v.4
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    • pp.85-95
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    • 1965
  • This report is to observe the amino acid composition at silk fibroins among varieties by using paper chromatography method and to study how they may concern with the genetical problem and the biochemical synthesis of the fibroin in silkgland. The results reached are as follows: 1. The amino acid composition of silk fibroin among varieties were observed as similar with each other on the paper chromatogram pictures. 2. In spite of the results of the amino acid composition analysis was similar with other workers results, there was some difference among varieties in numerical analysis. 3. The amino acid composition was considered to concern somewhat with the genetic problem of silkworm. 4. Some nutritive amino acids are considered to related with the worm growth more than the silk fibroin. 5. The paper chromatography of this report showed an unknown amino acid which was not found before in specific species silk fibroin and it is imagined as Hydroxyproline in view of other worker's graphy result. 6. The biochemical synthesis mechanism of silk fibroin in the silk gland, neither Stepwise process nor Template, could stand with the results of this paper satisfactory one side.

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