• Title/Summary/Keyword: reversed phase

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Involvement of $Cu^{++}$-Catalyzed Peroxidation in Degradation of Collagen and Protective Mechanism of Sodium Salicylate on this Peroxidative Reaction ($Cu^{++}$ 촉매작용에 의한 과산화 현상이 Collagen 손상에 관여함과 Sodium Salicylate에 의한 보호 작용)

  • Kim, Yong-Sik
    • The Korean Journal of Pharmacology
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    • v.23 no.1
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    • pp.25-31
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    • 1987
  • The present study examines firstly, the inhibition of collagen gelation to explore the possible involvement of $Cu^{++}$-catalyzed peroxidation in rheumatoid arthritis and secondly, the effect of sodium salicylate on this peroxidative reaction to provide a possible explanation for its mechanism of anti-inflammatory action. Incubation of collagen obtained from rat skin with $Cu^{++}$ and $H_2O_2$ resulted in the inhibition of gelation in terms of maximal turbidity and lag phase, but either $Cu^{++}$ or $H_2O_2$ alone essentially gave no effect in the collagen gelation. In the presence of sodium salicylate the inhibited gelation of collagen induced by $Cu^{++}$ and $H_2O_2$ was reversed with the dependency of the concentration of sodium salicylate. Moreover, the rate of $H_2O_2$ decomposition by $Cu^{++}$ was accelerated by sodium salicylate and this decomposition of $H_2O_2$ was found to be saturable in terms of concentration of this drugs. Thus it can be expected that $Cu^{++}$ -catalyzed peroxidation attacks collagen resulting in change of structural or functional integrity of collagen, and sodium salicylate may act on this peroxidative process, possibly through the enhancement of catalatic action of $Cu^{++}$. From these results $Cu^{++}$-catalyzed peroxidation can be in part responsible for degradation of joint tissue in rheumatoid arthritis and sodium salicylate may exert its anti-inflammatory action by this peroxidative reaction.

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Analysis of Cyclic Adenosine Monophosphate (cAMP) Separation via RP-HPLC (reversed-phase high-performance liquid chromatography) by the Moment Method and the van Deemter Equation (역상 크로마토그래피에서 모멘트 방법과 van Deemter 식을 이용한 고리형 아데노신 일인산의 분리특성 연구)

  • Lee, Il Song;Ko, Kwan Young;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.53 no.6
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    • pp.723-729
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    • 2015
  • The moment analysis of cyclic adenosine monophosphate (cAMP) was performed using chromatograms that were obtained with the pulse input method from an octadecyl silica (ODS) high-performance liquid chromatography (HPLC) column. The general rate (GR) model was employed to calculate the first absolute moment and the second central moment. Three important coefficients for moment analysis, which are molecular diffusivity ($D_m$), external mass transfer coefficient ($k_f$), and intra-particle diffusivity ($D_e$), were estimated by the Wilke-Chang equation, Wilson-Geankoplis equation, and comparing van Deemter equation to theoretical plate number equation, respectively. Experiments were conducted by various conditions of flow rates, methanol volume ratio of the mobile phase, and solute concentration. After the moment analysis, results were organized by van Deemter plots. Also van Deemter coefficients were compared each other to effect $H_{ax}$, $H_f$, and $H_d$ on height equivalent to a theoretical plate (HETP, $H_{total}$). The value of intraparticle diffusion ($H_d$) was the primary factor which makes for HETP whereas external mass transfer ($H_f$) was disregardable factor.

A study on the residues of fluoroquinolones in hen's eggs consumed in Incheon Metropolitan city (인천광역시에서 유통되는 계란의 플루오로퀴놀론계 항균물질 잔류 실태 조사)

  • Park, Eun-Jeong;Lim, Ji-Huen;Lee, Sung-Mo;Im, Jeong-Soo;Oh, Dae-Kyu
    • Korean Journal of Veterinary Service
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    • v.31 no.3
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    • pp.385-395
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    • 2008
  • This study was carried out to investigate the residue level of fluoroquinolones in hen's general eggs and specific eggs by microbiological assay method and high performance liquid chromatography (HPLC) method. HPLC separation was carried out by reversed phase chromatography on a Symmetry $C_{18}$ (250${\times}$4.6 mm, $5{\mu}m$ particle size) with a phase composed of distilled water (containing 0.4% triethylamine and phosphoric acid) : Methanol (780 : 220, v/v), pumped isocratically at a flow rate of 1.0ml/min. A fluorescence detector was utilized with an excitation wavelength of 278nm and an emission wavelength of 456nm. The calibration curves were linear $({\gamma}^2{\geq}0.999)$ over a concentration range of $0.025{\sim}0.4{\mu}g/ml$. Average recoveries of the five fluoroquinolones in whole eggs at fortified levels of $0.05{\sim}0.2{\mu}g/g$ were ranged mean $78.1{\sim}91.7%$ and low coefficient of variation was less than 10% for all analysed samples. The limits of detection and limits of quantification for whole eggs were $1.2{\sim}6.0ng/g$ and $2.3{\sim}9.1ng/g$, respectively. Only one hen's general eggfrom chicken farm in Incheon was detected with the residual fluoroquinolones (Microbiological assay method; 1 of 47 general eggs) ; the range of residual concentration enrofloxacin was 0.12ppm. Those in food stores were detected with the residual fluoroquinolones (Microbiological assay method; 4 of 88 general eggs) ; the ranges of residual concentration enrofloxacin were $0.15{\sim}2.2 ppm$, ciprofloxacin $0.01{\sim}0.06ppm$, and hen's specific eggs (40) in food stores were not detected. For the microbiological assay method of fluoroquinolones in hen's eggs, as the results of comparative analysis, the disc diffusion method with E coli may be a little highly detected for the residual fluoroquinolones.

Quantitative Determination and Stability of Curcuminoid Pigments from Turmeric (Curcuma longa L.) Root (강황 (Curcuma longa L.) 색소의 정량 및 안정성)

  • Kim Kwan-Su;Choung Myoung-Gun;Park Si-Hyung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.spc1
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    • pp.211-215
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    • 2005
  • Three curcuminoids [curcumin (CUR), demethoxycurcumin(DEM), bisdemethoxycurcu in (BIS)] are major yellow pigments in turmleric (Curcuma longa L.) root. Contents of curcuminoids in turmeric roots collected from 6 locations were analyzed using, high performance liquid chromatography (HPLC) equipped with reversed-phase column, an UV-Vis detector at 420nm, and eluted with a mixture of acetonitrile: $0.1\%$ acetic acid in water (50 : 50, v/v) as mobile phase. The stability of curcuminoid pigments in $80\%$ methanol extract solution were investigated during storage in a freezer at $-20^{\circ}C$, room temperature in the dark, and light condition. Calibration curves for the determination of curcuminoids were made with significant linearity $(r^2=0.999**)$. Average content of total curcuminoids was 171.5 mg/g, with 91.6 mg/g of CUR, 56.9 mg/g of DEM, and 23.0 mg/g of BIS. Amount of curcuminoids during storage in a freezer was almost not changed while those in room temperature wert reduced and rapid degradation appeared after 60 days. Within 90 days, about $50\%$ curcuminoid decreased in the dark and about $70\%$ in the light condition, indicating the decomposition of curcuminoid pigments followed under light and heat.

Retention Behavior and Separation of Phenol Derivatives through Cyclodextrin Complexes in Reversed-Phase Liquid Chromatography (역상 액체 크로마토그래피에서 씨클로덱스트린 착물을 이용한 페놀유도체들의 머무름 거동 및 분리)

  • Moon, Young-Ja;Kang, Sam-Woo
    • Analytical Science and Technology
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    • v.11 no.3
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    • pp.179-188
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    • 1998
  • The capacity factor of fifteen phenol derivatives was determined with respect to the concentration of ${\alpha}$- or ${\beta}$-cyclodextrin [CD], the type as well as the content of organic solvent in the mobile phase, and the temperature. The effect of the inclusion complex formation between solutes and ${\alpha}$- or ${\beta}$-cyclodextrin on their retention and selectivity has been investigated. The inclusion effect of ${\beta}$-cyclodextrin was the most effective in aqueous methanol, whereas only a poor effect was observed in aqueous tetrahydrofuran and aqueous acetonitrile. A plot of the reciprocal of the capacity factor against $[CD]_T$ gives a straight line and the dissociation constant, $K_D$ of the inclusion complex can be calculated from the slope. It was possible to estimate the $k_D$ values in 100% water from a linear plot of $pK_D$ vs. water content in the solution by extrapolation. The separation factor, ${\alpha}$, of two compounds has been found to be affected not only by the $[CD]_T$ but also by their $K_D$ values. Under optimum conditions, some mixtures of phenol derivatives were able to separate successfully.

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Purification of Two Novel Antimicrobial Peptides from Pyloric Caeca of the Starfish Asterina pectinifera (별불가사리 Asterina pectinifera의 유문맹낭 추출물로부터 새로운 2종류의 항균활성 펩타이드의 정제)

  • Go, Hye-Jin;Bae, Yun Jung;Park, Nam Gyu
    • Journal of Life Science
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    • v.24 no.8
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    • pp.860-864
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    • 2014
  • PAP-1, a novel antimicrobial peptide isolated from pyloric caeca extract of the starfish Asterina pectinifera was purified and characterized. First, the acidified pyloric caeca extract was put through Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient. Among the eluents, RM 60 (retained materials at 60% methanol) showed good antimicrobial activity against Bacillus subtilis and Escherichia coli D31 and was purified in C18 reversed-phase and ion-exchange high-performance liquid chromatography columns. The purification steps yielded two novel peptides showing strong antimicrobial activities. These peptides were named pyloric caeca A. pectinifera peptide 1 and 2 (PAP-1 and PAP-2). For the characterization of the purified peptides, the molecular weights and amino acid sequences were determined by MALDI-TOF MS and Edman degradation. The molecular weights of PAP-1 and PAP-2 were about 2951.54 Da and 2980.15 Da respectively. The amino acid sequences of PAP-1 and PAP-2 were partially determined: AIQNAGES and AIQNAAES, respectively. PAP-2 is an isoform of PAP-1, differing merely by a single residue at position 6 (glycine or alanine). The comparison of the N-terminal amino acid sequences and molecular weights of the peptides with those of other known antimicrobial peptides revealed that PAP-1 and PAP-2 have no homology with any known peptides. These findings suggest that PAP-1 and PAP-2 play a significant role in the innate defense system of starfish pyloric caeca.

Detection of Undeclared Betamethasone Derivatives in Cosmetic Products Labeled to Contain Zinc Pyrithione as the Active Ingredient (아연피리치온을 유효성분으로 표기한 화장품류에서 미표기 성분인 베타메타손 유도체의 검출)

  • Lee, Jeong-Pyo;Park, Sung-Hwan;Yang, Seong-Jun;Kim, Sun-Mi;Son, Kyung-Hun;Yun, Mi-Ok;Choi, Sang-Sook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.11-17
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    • 2009
  • Betamethasone propionate, an anti-inflammatory glucocorticosteroid, was detected in cosmetics with no indication on the label of this compound as an ingredient. The product was formulated as a topical spray or shampoo and labeled to contain zinc pyrithione as the active ingredient. A thin-layer chromatographic analysis was carried out on silica gel plates to provide a first indication about the presence of a compound with steroid structure and reactivity; then high-performance liquid chromatography (HPLC) separation allowed the identification of the corticosteroid agent and its quantification. To identify the corticosteroid agent from these commercial samples we collected the fractions suspected to have ketol steroids by prep HPLC and identified the compound as betamethasone propionate by NMR and MS spectrometry. Then we synthesized the standard for the betamethasone 17-propionate and 21-propionate and quantitate the corticosteroids from the sample by HPLC with that standards. By this method we identified the corticosteroid compounds from some commercial cosmetics such as zinc pyrithione sprays. The finding of betamethasone propionate in the products was shown by comparison to an authenticated standard of betamethasone propionate by retention time on reverse-phase HPLC. Two of the tested products contained betamethasone propionate at the levels of 0.005 ${\sim}$ 0.02% and the others were free of betamethasone propionate.

Molecular Mechanism of the Antiproliferative Effect by Ginseng Panaxynol on a Human Malignant Melanoma Cell Line, SK-MEL-1 (인체 흑색종 세포주 SK-MEL-1에 대한 인삼 panaxynol의 항증식 효과 기전)

  • Cho Hongkeun;Yu Su-Jin;Roh Joo Young;;Hwang Woo-Ik;Sohn Jeongwon
    • Journal of Ginseng Research
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    • v.23 no.3 s.55
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    • pp.190-197
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    • 1999
  • In this study, the molecular mechanism of the growth inhibitory effect of panaxynol was investigated in a human malignant melanoma cell line, SK-MEL-l. In the cell cycle analysis, panaxynol arrested cell cycle progression of SK-MEL-I at the G1 phase. Immunoblot analysis demonstrated that panaxynol increased $p21^{WAF1}$ and decreased cdc2 expression. Protein levels of pl6, p27, E2F-1, Rb, and p53 were not changed. Thus, the changes in expression levels of $p21^{WAF1}$ and cdc2 apparently mediate the cell cycle arrest caused by panaxynol. In addition, cycloheximide (CHX) partially reversed the growth inhibition by panaxynol, which suggested that new protein synthesis was required. On the other hand, LLnL, a proteasome inhibitor, increased antiproliferative effect of panaxynol. This may be due to stabilization of the protein(s) responsible for the growth inhibition such as $p21^{WAF1}$. In summary, these results demonstrate that panaxynol inhibits proliferation of SK-MEL-I by inducing cell cycle arrest at the G1 phase and the inhibitory effect is mediated by the increased level of $p21^{WAF1}$ as well as decreased cdc2 expression.

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Analysis and Enzymatic Production of Structured Lipids Containing DHA Using a Stirred-Batch Type Reactor (회분식 반응기를 이용한 DHA 함유 재구성지질의 효소적 합성 및 이화학적 특성 분석)

  • Kim, Nam-Sook;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.7
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    • pp.1052-1058
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    • 2005
  • Structured lipid (SL) was synthesized by enzymatic interesterification with algae oil containing docosahexaenoic acid (DHA) and soybean oil in the stirred-batch type reactor. The reaction was performed for 15hr at $65^{\circ}C$ with 300 rpm catalyzed by sn-1,3 specific Lipozyme RM 1M lipase from Rhizomucor miehei ($11\%$ by weight of total substrates) in the absent organic solvent. SL contained $87.1\;area\%$ triacylglycerol (TAG), $12.1\;area\%$ diacylglycerol (DAG), $0.6\;area\%$ monoacylglycerol (MAG), and $0.2\;area\%$ free fatty acid (FFA). Major fatty acid profiles of SL were DHA $(15.7\;mol\%)$, linoleic $(31.1\;mol\%)$, palmitic $(20.2\;mol\%)$, oleic $(13.5\;mol\%)$ and eicosapentaenoic acid $(EPA,\;6.6 mol\%)$. SL contained the newly synthesized several peaks. Iodine and saponification of SL were 206.7 and 183.8. SL color showed darker and redder than soybean oil, and appeared the most yellowish color among SL, soybean, and algae oil.

Study on the Positional Distribution of Fatty Acids, and Triacylglycerol Separation, of Seed Oils (종실유(seeds oil)의 위치별 지방산 및 트리아실글리세롤의 조성 연구)

  • Moon, Jun-Hee;Hwang, Yun-Ik;Lee, Ki-Teak
    • Food Science and Preservation
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    • v.16 no.5
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    • pp.726-733
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    • 2009
  • Six types of oil were extracted from pomegranate seed, mung bean, pepper seed, safflower seed, seeds of Cassia tora Linnaeus, and perilla seed. The extracted seed oils were analyzed for total and positional fatty acid composition, triacylglycerol (TAG) level, and tocopherol content. Crude fat levels measured by the Folch method were 21.64% in perilla seed, 13.85% in safflower seed, 9.60% in pepper seed, 8.85% in pomegranate seed, 2.25% in mung bean, and 2.00% in C. tora,respectively (all w/w). Linoleic acid (C18:2) was the most abundant fatty acid at the sn-2 position of triacylglycerols (TAGs), ranging from 15.99-88.3 wt%. The composition of TAGs was analyzed by reverse-phase HPLC, and TAGs of seed oils showed partition numbers of 36-48. The highest content (377.74 mg/100 g) of total tocopherol was found in pomegranate seed whereas the total tocopherol content of mung bean, C. tora, pepper seed, perilla seed, and safflower seed were 141.16, 107.23, 33.88, 30.05, and 29.80 mg/100 g, respectively.