• Title/Summary/Keyword: reversed-phase HPLC

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Screening and Purification of an Antimicrobial Peptide from the Gill of the Manila Clam Ruditapes philippinarum (바지락(Ruditapes philippinarum) 아가미로부터 항균 펩타이드의 탐색 및 정제)

  • Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.2
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    • pp.137-145
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    • 2016
  • This study screened the biological activity of an acidified gill extract of the Manila clam Ruditapes philippinarum including antimicrobial, hemolytic, membrane permeabilization, and DNA-binding activity, and purified the antimicrobial material. The acidified gill extract showed potent antimicrobial activity against Bacillus subtilis and Escherichia coli without significant hemolytic activity, but showed no membrane permeabilization or DNA-binding ability. An antimicrobial material was purified from the acidified gill extract using C18 reversed-phase and cation-exchange high-performance liquid chromatography (HPLC). Treatment of the purified material with trypsin completely abolished all of the antibacterial activity against Bacillus subtilis, suggesting that the purified material is a proteinaceous antibiotic. The molecular weight of the purified material was 2571.9 Da, but no primary structural information was obtained due to N-terminal blocking. A future study should confirm the primary structure. Our results suggest that the Manila clam gill contains proteinaceous antibiotics that have a role in first-line defense. This information could be used to better understand the Manila clam innate immune system.

Characterization of β-Secretase Inhibitory Peptide Purified from Blackfin flounder (Glyptocephalus stelleri) Protein Hydrolysate

  • Lee, Jung Kwon;Kim, Sung Rae;Byun, Hee-Guk
    • Journal of Marine Bioscience and Biotechnology
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    • v.10 no.1
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    • pp.1-8
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    • 2018
  • The objective of this study was to purify and characterize the ${\beta}-secretase$ inhibitor from enzymatic hydrolysates of blackfin flounder muscle, for development of a novel anti-dementia agent that may be used in the drug or functional food industries. ${\beta}-secretase$ inhibitory peptide was purified from various enzymatic hydrolysates of blackfin flounder muscle. Among six enzymatic hydrolysates, the Alcalase hydrolysate revealed highest ${\beta}-secretase$ inhibitory activity. Consecutive purification of the blackfin flounder muscle hydrolysate using Sephadex G-25 column chromatography and octadecylsilane C18 reversed phase HPLC techniques were used to isolate a potent ${\beta}-secretase$ inhibitory peptide composed of 5 amino acids, Leu-Thr-Gln-Asp-Trp (MW: 526.7 Da). The $IC_{50}$ value of purified ${\beta}-secretase$ inhibitory peptide was $126.93{\mu}M$. Results of this study suggest that peptides derived from blackfin flounder muscle may be beneficial as anti-dementia compounds in functional foods or as pharmaceuticals.

Comparison of the Scrophulariae Radix of Gyeongbuk's with that of Chinese: Content of Harpagoside and Inhibitory Effect of Nitric Oxide (경북산 현삼과 중국산 현삼의 비교: Harpagoside 함량 및 Nitric Oxide 저해활성)

  • Zhang, Hai-Yan;Kim, Geum-Jin;Hwang, In-Hyun;Cho, Hi-Jae;Lee, Seung-Ho;Na, Min-Kyun
    • Korean Journal of Pharmacognosy
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    • v.42 no.1
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    • pp.15-21
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    • 2011
  • Scrophulariae Radix (Scrophulariaceae) has been traditionally used to treat pharyngitis, laryngitis, tonsillitis, fever, and edema. To compare the quality of the Scrophulariae Radix produced from Gyeongbuk with that from China, a quantitative analysis on the major component, E-harpagoside, was performed by reversed-phase HPLC, and a relative inhibitory activity of nitric oxide (NO) was assayed in the Raw 264.7 cells. Using a Luna $C_{18}$(2) column, we carried out quantitative analysis with a gradient of 0.1% phosphoric acid in water and acetonitrile. Our results demonstrated that the content of E-harpagoside in Gyeongbuk's Scrophulariae Radix were three times higher than that of Chinese. Moreover, the inhibitory activity of Scrophulariae Radix of Gyeongbuk's in the lipopolysaccharide (LPS)-induced NO production was higher than that of Chinese.

Isolation and Purification of Neuropeptides from the Tube Feet of the Starfish Asterias amurensis (아무르 불가사리(Asterias amurensis)의 관족으로부터 신경성 펩타이드의 분리 및 정제)

  • Jo, Mi Jeong;Go, Hye-Jin;Kim, Gun-Do;Park, Nam Gyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.2
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    • pp.129-134
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    • 2014
  • Two neuropeptides were purified from the acidified tube feet extract of the starfish Asterias amurensis by C18 reversed phase and size-exclusion high-performance liquid chromatography (HPLC). The tube feet extract and the purified peptides (AST-I and AST-II) showed potent contractile activity on dorsal retractor muscle (DRM) of the starfish Asterina pectinifera and intestine (smooth muscle) of the panther puffer Takifugu pardalis. Treatment of the purified peptides with dithiothreitol (DTT) for 60 min at $37^{\circ}C$ significantly altered their retention times, suggesting that these compounds contained disulfide bonds. The molecular weights of AST-I and AST-II were determined to be 2064.2 Da and 6137.2 Da, respectively, by MALDI-TOF mass spectrometry.

Antioxidant Activities of Acidic Ethanol Extract and the Anthocyanin Rich Fraction from Aronia melanocarpa (아로니아 추출물 및 아로니아 안토시아닌분획의 항산화 활성 효과)

  • Lim, Jung-Dae;Cha, Hwan-Soo;Choung, Myoung-Gun;Choi, Ri-Na;Choi, Duck-Joo;Youn, Aye-Ree
    • Korean journal of food and cookery science
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    • v.30 no.5
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    • pp.573-578
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    • 2014
  • The objective of this study was to investigate the antioxidant activities and anthocyanin profiles in the anthocyanin rich fraction (ARFAM) of Aronia melanocarpa, which are considered functional substances and are available as food coloring agents in Korea. Anthocyanins were identified by reversed-phase C18 column chromatography and HPLC-DAD-ESI/MS analysis. The antioxidative activity of the acidic ethanol extract (AME) and the anthocyanin-rich fraction (ARFAM) was determined by scavenging of the diphenylpicrylhydrazyl (DPPH) radical, the hydroxy radical, and the superoxide anion in addition to reducing power using a commercial antioxidant as a reference.

High-performance liquid chromatographic assay and oral pharmacokinetics of new anti-HIV uracil derivatives, KR-V analogues, in rats

  • Lee, Youngmi;Heeok Shim;Seoungryong Yu;Hochul Shin;Moonkoo Chung;Jungkoo Roh
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.123-123
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    • 1998
  • A number of uracil derivatives have been developed as anti-AIDS drugs having a mechanism of inhibiting cellular reverse transcriptase. A simple and rapid assay technique for recently synthesized KR-V analogues was developed using a high-performance liquid chromatography, and oral pharmacokinetics was examined for assessing their oral bioavailabilites. Plasma samples were analyzed by reversed-phase HPLC using an ODS column with an ultraviolet detection system. All the analogues were eluted within 12 min and the LOQ was 15-30 ng/$m\ell$. The extraction recoveries were higher than 85%, except KR-V1039, 1068 and 1720 having ester group. This chromatic method was well applied to the kinetic studies for KR-V analogues. Among 16 analogues tested in the present work, the 6 compounds including KR-V1123, 1122, 1784, 1783, 1736 and 1700 were found to be bioavailable for oral administration to rats.

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Facile Synthesis and Radioiodine Labeling of Hypericin

  • Kim, Sang-Wook;Park, Jeong-Hoon;Yang, Seung-Dae;Hur, Min-Goo;Kim, Yu-Seok;Chai, Jong-Seo;Kim, Young-Soon;Yu, Kook-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.25 no.8
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    • pp.1147-1150
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    • 2004
  • Hypericin (1,3,4,6,8,13-hexahydroxy-10,11-dimethylphenanthro[1,10,9,8-opqra]perylene-7,14-dione), an antidepressant which is also known to be a potent protein kinase C (PKC) inhibitor was synthesized as a precursor for radioiodine labeling via two step reactions. Malignant glioma cells express higher PKC activity compared to untransformed glial cell. Here we report the synthesis and radioiodine labeling of hypericin as a potential brain tumor imaging radiopharmaceutical. The reference compound, 2-iodohypericin, and its radiolabelled analogues, 2-[$^{123}I$]iodohypericin and 2-[$^{124}I$]iodohypericin have been prepared by the reaction of hypericin with NaI or [$^{123}I$]NaI or [$^{124}I$]NaI. The labeling yield was 60-65% for each analogue and the optimal reaction time was 10 min. The purification and isolation of the labelled products were achieved by a reversed-phase HPLC.

Gamnamoside, a Phenylpropanoid Glycoside from Persimmon Leaves (Diospyros kaki) with an Inhibitory Effect against an Alcohol Metabolizing Enzyme

  • Varughese, Titto;Rahaman, Mozahidur;Kim, No-Soo;Cho, Soon-Chang;Moon, Surk-Sik
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1035-1038
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    • 2009
  • Phytochemical investigation of the methanolic extract of Diospyros kaki leaves led to the isolation of osmanthuside H (1) and a new phenol glycoside, named gamnamoside [4-(3-hydroxypropyl)-2-methoxyphenol $\beta$-D-apiofuranosyl( 1 $\rightarrow$ 6)$\beta$-D-glucopyranoside] (2) along with (-) catechin (3) through a series of reversed phase column chromatography and preparative C18 HPLC. The structures of the isolates were determined by spectroscopic methods including IR, UV, HRTOFMS, and 2D NMR. Compounds 1, 2, and 3, showed good inhibitory activities ($IC_{50}$) of 175.4, 94.4, and 126.6 ${\mu}g/mL$ respectively, whereas a reversible ADH inhibitor, 4-methylpyrazole, showed the $IC_{50}$ of 326.6 ${\mu}g/mL$ against alcohol dehydrogenase (ADH).

Effects of Climatic Factors and Tapping Date on Yield and Quality of Lactree (Rhus verniciflua) Sap (기상요인과 채취시기가 옻나무 칠액채취량 및 칠액의 質에 미치는 영향)

  • 김만조
    • Korean Journal of Plant Resources
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    • v.11 no.1
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    • pp.70-79
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    • 1998
  • This study was conducted to understand the effects of weather and tapping date on yield and quality of lactree(Rhus verniciflua) sap yield showed a significant positive correlation with the minimum temperature of one day before sap collection at 1% level and with theminimum humidity of theday of sap collection at 5% level. However, the differences between the maximum and the minimum temperatures and humidities of the day of sap collection were negatively correlated with the sap yield at 5% level. Multiple regression analysis indicated that the minimum temperature of one day bofore sap collection and the minumum humidity of the day sap collection were important factors for increasing sap yield. The high sap yield of lactree by Japanese tapping method was recorded during mid-July and early August. Seasonal variation in lactree sap constituents was observed. The sap collected on 15th of August contained the highest urushiol content (68.3%) and the lowest water content resulting in high quality of lactree sap. By reversed-phase HPLC analysis, fove urushiol components were separated from each other depending on the number of doulbe bonds in the side-chain , and seaxonal variation of urushiol composition was noticed. The 3-C15 triene content fo the sap collected on 5th of July was the highest(77.56%) indicating the major component of urushiol which affects lactree sap quality.

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Inhibitory Effect of Kaurane Type Diterpenoids from Acanthopanax koreanum on TNF-$\alpha$ Secretion from Trypsin-Stimulated HMC-1 Cells

  • Cai, Xing-Fu;Shen, Guanghai;Dat, Nguyen-Tien;Kang, Ok-Hwa;Lee, Young-Mi;Lee, Jung-Joon;Kim, Young-Ho
    • Archives of Pharmacal Research
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    • v.26 no.9
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    • pp.731-734
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    • 2003
  • Five known kaurane type diterpenoids, 16$\alpha$H, 17-isovaleryloxy-ent-kauran-19-oic acid (1), 16$\alpha$-hydroxy-17-isovaleryloxy-ent-kauran-19-oic acid (2), paniculoside-IV (3), 16$\alpha$-hydroxy-ent-kauran-19-oic acid (4), and ent-kaur-16-en-19-oic acid (5) were isolated from the root of Acanthopanax koreanum by repeated column chromatography and reversed phase preparative HPLC. The structures of these compounds were established from physicochemical and spectral data. Among the isolated compounds 16$\alpha$H, 17-isovaleryloxy-ent-kauran-19-oic acid (1) showed potent inhibitory activity ($IC_50$ value, 16.2 $\mu$ M) on TNF-$\alpha$ secretion from HMC-1, a trypsin-stimulated human leukemic mast cell line.