• Title/Summary/Keyword: RP HPLC

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Antimicrobial Constituents from Fruits of Ailanthus Altissima SWINGLE

  • Zhao Chun-Chao;Shao Jian-Hua;Li Xian;Xu Jing;Zhang Peng
    • Archives of Pharmacal Research
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    • v.28 no.10
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    • pp.1147-1151
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    • 2005
  • A new naturally occurring sterol, compound 5, and six known stigmasterols were isolated from fruits of Ailanthus altissima Swingle by repeated column chromatography and RP-HPLC. Their structures were identified as, 5${\alpha}$-stigmastane-3,6-dione (1), 3${\beta}$-hydroxystigmast-5-en-7-one (2), stigmast-5-ene-3${\beta}$, 7${\alpha}$-diol (3), 6${\alpha}$-hydroxystigmast-4-en-3-one (4), 5${\alpha}$-stigmastane-3${\beta}$, 6${\beta}$-diol (5), stigmast-4-ene-3${\beta}$, 6${\alpha}$-diol (6), stigmast-5-ene-3${\beta}$, 7${\alpha}$, 20$\xi$-triol (7) by spectral analysis and comparison with the published data. These compounds have not been reported from genus Ailanthus, whereas compound 7 was identified by NMR for the first time. In addition, the $95\%$ ethanol extract and compounds from the fruits of Ailanthus altissima SWINGLE were assayed for in vitro antimicrobial activity. The extract was potent active against the assayed bacteria while compounds 3 and 7 exhibited moderate activity.

An Antioxidant Homo-Flavoyadorinin-B from Korean Mistletoe (Viscum album var. coloratum) (한국산 겨우살이(Viscum album var. coloratum)로부터 분리한 homo-flavoyadorinin-B의 항산화 활성)

  • Choi, Seung-Young;Chung, Shin-Kyo;Kim, Suk-Kyung;Yoo, Yung-Choon;Lee, Kyung-Bok;Kim, Jong-Bae;Kim, Ja-Young;Song, Kyung-Sik
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.279-282
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    • 2004
  • An antioxidant was isolated from Korean mistletoe (Viscum album var. coloratum) by consecutive purification using silica gel, Sephadex LH-20, and RP-HPLC. The active principle was identified as homo-flavoyadorinin-B (3',7-dimethoxyluteolin-4'-O-[apiosyl $(1{\rightarrow}2)$ glucoside]) by spectral analyses. It inhibited 74.6% of hydroxyl radical and 30.6% of superoxide anion radical at 0.01 mM; however, th~compound did not show any scavenging activity against hydrogen peroxide radical. At 0.1 mM, above compound scavenged superoxide anion radical about twice as effective as positive controls, BHT and ${\alpha}-tocopherol$. Radical scavenging activities of homo-flavoyadorinin-B on DPPH, hydroxyl, and hydrogen peroxide radicals were almost same with those of positive controls.

Dyeing Behavior of Silk Dyed with Indigo Leaf Powder Using Reduction and Nonreduction Dyeing and Its Relationship with the Amount of Indigotin and Indirubin Adsorbed in Silk

  • Yoo, Wansong;Ahn, Cheunsoon
    • Journal of the Korean Society of Clothing and Textiles
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    • v.43 no.5
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    • pp.753-767
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    • 2019
  • Dyeing behavior of indigo leaf powder was examined in regards to the effect of the amount of pigments on color and dye adsorption for silk dyed by reduction and nonreduction dyeing. The amount of indigotin and indirubin pigments adsorbed in dyed silk was examined by HPLC-DAD analysis. The color of dyed silk showed 7.7BG - 2.7B hue when silk was dyed at $50^{\circ}C$, and 3.5G - 4.9BG when dyed at $70^{\circ}C$. Blue ($b^*$) and green ($a^*$) color decreased as the pH of dyebath increased. When silk was dyed using nonreduction, R (red) and RP (red purple) hue and R hue was more apparent in samples dyed at $90^{\circ}C$. In reduction dyeing, amount of indigotin detected from silk exceeded the amount that was initially contained in the input dye. The amount of indirubin was lower than indirubin that was initially in the powder. In nonreduction dyeing, silk showed a higher amount of indirubin adsorption compared to silk dyed by reduction. The amount of indigotin adsorbed in silk was lower than the amount initially contained in the input dye. The amount of indigotin and indirubin adsorption was primarily dependent upon the dyeing method-reduction or nonreduction along with dyeing temperature and the pH of dyebath.

Relationship of Phenolic Compounds and Free-radical Scavenging Activity in Black and Red Rice Extract

  • Hu, Gao-Sheng;Xu, Kai-Xiu;Jeong, Soon Jae;Kim, Doh-Hoon
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.129-138
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    • 2010
  • Total phenolic content, total anthocyanin content and antioxidant activity were analyzed from rice samples collected in Korea, Japan and China. The results showed that the total phenolic content and free-radical scavenging activity differed significantly in these rice lines. The correlation between content and activity was subsequently investigated. The results showed that in black rice, anthocyanin was the major phenolic component and that both phenolic content and anthocyanin content were closely correlated with free-radical scavenging activity. Reverse Phase High Performance Liquid Chromatography (RP-HPLC) data showed that cyanidin-3-O-glucoside and peonidin-3-O-glucoside composed about 90% of the total anthocyanin content in black rice and in red rice. In the red rice extract, the total phenolic content produced a high correlation coefficient with antioxidant activity but correlated very poorly with the total anthocyanin content. The $OD_{458}$ and the $OD_{500}$ values which represent the proanthocyanidin content of the rice extract, produced high correlation coefficients with antioxidant activity and total phenolic content. These results suggest that the $OD_{458}$ and the $OD_{500}$ values can be used to evaluate the quality of red rice. In addition, based on the data obtained, a competitive accumulation model of anthocyanin and proanthocyanidin in black and red rice was proposed.

ELUTION OF RESIDUAL MONOMER ACCORDING TO VARIOUS LIGHT SOURCES AND CURING TIME ON THE POLYMERIZATION OF PHOTOACTIVATED PIT AND FISSURE SEALANTS (광중합 광원의 종류와 조사시간에 따른 치면열구전색제의 미반응 모노머 용출)

  • Oh, You-Hyang;Park, Yoon-Kyung;Lee, Nan-Young;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.3
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    • pp.421-430
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    • 2004
  • The purpose of this study was to measure and compare the amount of unreacted TEGDMA from pit and fissure sealants cured with three different light sources; conventional halogen light curing unit, plasma arc light curing unit and argon laser. The specimens were eluted in distilled water for different time intervals. The time-related release of TEGDMA were analyzed by reverse-phase high performance liquid chromatography(HPLC). The result of present study can be summarized as follows: 1. The time-related release of TEGDMA decreased with increasing curing time in conventional halogen light, however, that not statistically significant difference(p>0.05). 2. The elution from the specimens cured for 6 and 9 seconds with plasma arc light was similar results corresponding with the time-related TEGBMA release, and was significantly lower than that cured for 3 seconds(p<0.05). 3. The elution of TEGDMA from the specimens cured with argon laser was significantly higher than that cured with halogen and plasma arc light(p<0.05). 4. The elution of TEGDMA from under recommended time of three different light sources were showed to be no statistically significant difference(p>0.05). 5. In time-related release of TEGDMA from recommended time of each light sources, the results correspond to 40 seconds of halogen light and 6 seconds of plasma arc light were similar(p>0.05). 6. The elution of TEGDMA, from over recommended time of three different light sources were showed to be no statistically significant difference(p>0.05). In this study, I suggest that curing time of plasma arc light is 6 and/or 9 seconds in the field of clinical pediatric dentistry claiming its effectiveness in optimal polymerization and reduced chair time.

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Enzymatic Interesterification and Melting Characteristic for Asymmetric 1,2-Distearoyl-3-Oleoyl-rac-Glycerol Triacylglycerol Enriched Product (효소적 반응을 이용한 비대칭형 1,2-Distearoyl-3-Oleoyl-rac-Glycerol 혼합물의 생성 및 융점 특성)

  • Kim, Jin Young;Lee, Ki Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.93-101
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    • 2014
  • Asymmetric 1,2-distearoyl-3-oleoyl-rac-glycerol (SSO) triacylglycerol (TAG) is used as a cocoa butter replacer (CBR). In this study, it was produced by lipase-catalyzed interesterification of fully hydrogenated soybean oil (FHSBO) and oleic ethyl ester (OEE) in a batch type reactor at $75^{\circ}C$, 250 rpm. Different molar ratios (FHSBO : OEE=1:1, 1:2 and 1:3, w/w) and various reaction times (1, 2, 3, 4, and 5 hr) were also tested. The optimized condition for SSO was a FHSBO : OEE molar ratio of =1:1 at reaction times of 2, 3, 4, and 5 hr. Enzymatic synthesis generated SSO/SOS, as well as the other TAGs (e.g., PSO/POS, SOO/OSO, SSS), ethyl esters, monoacylglycerol (MAG), and diacylglycerol (DAG). After scale-up, fractionation by solvent (methanol and acetone) fractionation and column chromatography was applied. To reduce ethyl esters, high-melting TAGs (e.g., SSS), and SOO/OSO in reactants, solvent fractionation was applied. Using a silica gel column (sample : silica gel=2:1, wt%), MAG and DAG were removed at $25^{\circ}C$. The major fatty acid composition of the final products (with a high SSO/SOS content) was palmitic acid (C16:0, 10.9~12.9 area%), stearic acid (C18:0, 52.2~54.9 area%), and oleic acid (C18:1, 34.2~35.5 area%). In reversed-phase HPLC analysis, the major TAG species of the final product (FHSBO : OEE=1:1, 2 hr) were SSO/SOS (82.31 area%) and PSO/POS (14.51 area%). Based on the $[SS]^+$ : $[SO]^+$ ratio obtained by RP-HPLC/APCI-MS, the final product had a higher SSO (AAB type TAG) content than cocoa butter (CB). The solid fat index (SFI) of CB and the final product obtained were similar with a narrow melting point range around ~32 to $35^{\circ}C$.

Secondary Metabolites Production and Plant Growth Promotion by Pseudomonas chlororaphis and P. aurantiaca Strains Isolated from Cactus, Cotton, and Para Grass

  • Shahid, Izzah;Rizwan, Muhammad;Baig, Deeba Noreen;Saleem, Rahman Shahzaib;Malik, Kauser A.;Mehnaz, Samina
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.480-491
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    • 2017
  • Fluorescent pseudomonads have been isolated from halophytes, mesophytes, and xerophytes of Pakistan. Among these, eight isolates, GS-1, GS-3, GS-4, GS-6, GS-7, FS-2 (cactus), ARS-38 (cotton), and RP-4 (para grass), showed antifungal activity and were selected for detailed study. Based on biochemical tests and 16S rRNA gene sequences, these were identified as strains of P. chlororaphis subsp. chlororaphis and aurantiaca. Secondary metabolites of these strains were analyzed by LC-MS. Phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine, Cyclic Lipopeptide (white line-inducing principle (WLIP)), and lahorenoic acid A were detected in variable amounts in these strains. P. aurantiaca PB-St2 was used as a reference as it is known for the production of these compounds. The phzO and PCA genes were amplified to assure that production of these compounds is not an artifact. Indole acetic acid production was confirmed and quantified by HPLC. HCN and siderophore production by all strains was observed by plate assays. These strains did not solubilize phosphate, but five strains were positive for zinc solubilization. Wheat seedlings were inoculated with these strains to observe their effect on plant growth. P. aurantiaca strains PB-St2 and GS-6 and P. chlororaphis RP-4 significantly increased both root and shoot dry weights, as compared with uninoculated plants. However, P. aurantiaca strains FS-2 and ARS-38 significantly increased root and shoot dry weights, respectively. All strains except PB-St2 and ARS-38 significantly increased the root length. This is the first report of the isolation of P. aurantiaca from cotton and cactus, P. chlororaphis from para grass, WLIP and lahorenoic acid A production by P. chlororaphis, and zinc solubilization by P. chlororaphis and P. aurantiaca.

Purification of a Protease Produced by Bacillus subtilis PCA 20-3 Isolated from Korean Traditional Meju (전통 메주로부터 분리한 Bacillus subtilis PCA 20-3 유래 Protease 의 정제)

  • Lim, Seong-Il;Yoo, Jin-Young
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1635-1641
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    • 1999
  • Bacillus subtilis PCA20-3 was isolated from meju and was found to produce a protease. The strain produced the maximum amount of enzyme in the medium containing soytone (0.2%), soluble starch (2%), $(NH_4)_2SO_4\;(0.1%),\;CaCl_2(0.1%),\;yeast\;extract\;(0.01%),\;K_2HPO_4\;(0.1%),\;and\;KH_2PO_4\;(0.1%)$. Protease was first concentrated by ammonium sulfate (80% saturation, w/v) precipitation of culture supernatant. Then the enzyme was purified by column chromatography using CM Sephadex C-50. The collected proteins were rechromatographed using Sephadex G-100 gel filtration column. The fraction with protease active from Sephadex G-100 gel chromatography was found to be pure when examined by SDS-polyacrylamide gel electrophoresis and YMC-pak reverse phase chromatography. Specific activity, yield and purity were 76 U/mg. 2.7%, and 7.6 fold, respectively. The molecular weight of the enzyme was estimated to be 31.5 kDa by SDS-PAGE. The number of amino acids calculated from molecular weight was evaluated about 321 residues. N-terminal sequence of the enzyme was $Val^1-Pro^2-Tyr^3-Gly^4-Val^5-Ser^6-Gln^7-Gly^8-Lys^9-Ala^{10}$.

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Optimization of Enzymatic Synthesis Condition of Structured Lipids by Response Surface Methodology (반응표면분석에 의한 기능성 유지의 효소적 합성 조건 최적화)

  • Cho, Eun-Jin;Lee, Jong-Ho;Lee, Ki-Teak
    • Korean Journal of Food Science and Technology
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    • v.36 no.4
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    • pp.531-536
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    • 2004
  • Synthesis conditions were optimized using response surface methodology for producing structured lipids (SL) by interesterification of DHA-enriched algae oil derived from microalgae, Schizochytrium sp. and corn oil. Reaction was performed fer 24 hr at $55^{\circ}C$ catalyzed by immobilized lipase from Rhizomucor miehei (RM IM) in shaking water bath. Major fatty acids of SL were palmitic (21.70 mol%), oleic (20.20 mol%), and linoleic (27.34 mol%) acids, and DHA (15.06 mol%). To separate newly synthesized SL-triglycerides (TG) species, HPLC with evaporative light scatting detector (ELSD) was used. Production conditions were optimized using central composite design with reaction temperature $(35-75^{\circ}C,\;X_1)$, reaction time $(2-42\;hr,\;X_2)$, and enzyme concentration $(2-14%,\;X_3)$ as variables. When variables were $70.28^{\circ}C\;(X_1),\;28.74\;hr\;(X_2),\;and\;11.30%\;(X_3)$, maximum content of selected three peaks of synthesized SL-TG species was predicted as 6.97 area%.

Synthesis Characterization and Biodistribution of $^{99m}Tc$-Ethyl-3-Isocyanobutyrate as a New Myocardial Perfusion Agent (새로운 심관관류 영상 화합물로서 $^{99m}Tc$-Ethyl-3-Isocyano-butyrate의 합성, 표지 및 체내동태에 대한 연구)

  • Lee, Myung-Chul;Cho, Jung-Hyuck;Lee, Dong-Soo;Lim, Sang-Moo;Oh, Seung-Joon;Chung, Soo-Wook;Lee, Kyung-Han;Jeong, Jae-Min;Chung, June-Key;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.27 no.2
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    • pp.223-232
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    • 1993
  • Technetium labeled isonitrile analogues are widely used as myocardial perfusion imaging agents. We synthesized and characterized a new isonitrile compound, ethyl 3-isocyanobutyrate(EIB). Proton and $^{13}C$ NMR spectroscopy and thin layer chromatography with a $C_{18}$ coat was performed. EIB was easily labeled with $^{99m}TcO_4^-$- with sodium dithionite. The labeling efficiency measured by RP-HPLC was over 95%. The labeled product was stable with dilution in normal saline and with prolonged incubation at room temperature. There was no formation of secondary products or free $^{99m}TcO_4^-$. In vivo kinetics study of $^{99m}Tc$ (I) labeled EIB in rabbits showed adequate myocardial uptake, good contrast against lung background, and relatively rapid liver clearance. The heart to lung ratio was over 2.5 and the heart to liver ratio was approximately from 0.4 to 5 at 60 minutes post injection. Hepatic clearance of $^{99m}Tc-MIBI$ was faster ($t_{1/2}$=6 minutes) than that of $^{99m}Tc-MIBI$. In vivo kinetics observed in dog was similar to that in rabbit but there was faster gallbladder filling, and thus lower liver background. SPECT imaging of the canine myocardium showed favorable imaging characteristics. However, biodistribution in mice demonstrated a myocardial % injected dose/organ of less than 0.1%. This was thought to be due to interspecies difference in plasma esterase activity. In human plasma, $^{99m}Tc$ ( I ) labeled EIB was stable for at least 2 hours, without production of secondary products by HPLC. We conclude that ethyl 3-isocyanobutyrate may be a potential new myocardial perfusion imaging agent and deserves further investigation as to its usefulness for clinical use.

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