• Title/Summary/Keyword: reversed phase

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Identification of Proteins in Egg White Using Ion Exchange Cartridge and RP-HPLC (이온교환 카트리지와 RP-HPLC를 이용한 난백 단백질의 확인)

  • Kim, Hyun Moon;Kim, Ah Reum;Lee, Chang Soo;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.713-717
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    • 2012
  • Approximately forty proteins in egg white have been widely studied for their functional properties. To develop a procedure of separation for pure and non-altered proteins from egg white, purification study was conducted to isolate lysozyme, ovotransferrin, and ovalbumin. Ion exchange cartridge can selectively separate proteins from egg white, and reversed-phase HPLC (RP-HPLC) could identify separated proteins. Proteins in egg white were purified by HI trap ion exchange cartridge SP and Q with buffers pH 8.0 and 5.2. C18 column (Phenomenex, USA) was used for RP-HPLC analysis and isocratic mobile phase was used with acetonitrile (ACN)/distilled water (DW)/trifluoroacetic acid (TFA) in the ratio of 50/50/0.1. Comparing the retention times of standards in RP-HPLC experiments showed that ovotransferrin, ovalbumin, and lysozyme in egg white were eluted successively in the RP-HPLC column after the pretreatment in SP and Q ion exchange cartridges.

A Study on the Analysis of Polycyclic Aromatic Hydrocarbons by RPLC/DAD (I) (RPLC/DAD를 이용한 Polycyclic Aromatic Hydrocarbon류의 분석에 관한 연구(I))

  • Lee, Won;Hong, Jee-Eun;Park, Song-Ja;Pyo, Hee Soo
    • Analytical Science and Technology
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    • v.10 no.5
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    • pp.315-324
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    • 1997
  • The retention behaviors of 16 PAHs and 4 nitro-PAHs were studied with several parameters involved numbers of carbon atoms, F factor, aqueous solubility, L/B ratio, and numbers of interfering hydrogen atom pairs on the chemical structures of PAHs by using reversed-phase liquid chromatography/diode array detection method (RPLC/DAD) and gradient elution method. It was obtain that the log k' for most of PAHs with increasing the number of carbon and the F factor in their molecules. Chromatographic retention of PAH isomers and nitro-PAHs were examined with aqueous solubility, L/B ratio and number of interfering hydrogen atom pairs. As a result of comparison with these factors and retention times, it was found that those solutes having larger aqueous solubilities and greater L/B ratios were retained longer on stationary phase. This tendency was also occured in the molecules having the more number of interfering hydrogen atom pairs. Detection limits of PAHs which were obtained with three times measurements by RPLC/DAD were in the range of 100~500ng/mL and method detection limit(MDL) for water sample were in the range of 0.1~0.5ng/mL.

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HPLC Analysis and Pharmacokinetics of KAL-1120, a Novel Anti-inflammation Agent, in Rats (흰쥐에서 항염증제 후보물질 KAL-1120의 HPLC 분석 및 약물동태)

  • Shin, Dae-Hwan;Lee, Jung-Yeol;Park, Seong-Hyeok;Lee, Gyeong-Bok;Han, Kun;Chung, Youn-Bok
    • Journal of Pharmaceutical Investigation
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    • v.40 no.2
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    • pp.101-107
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    • 2010
  • A rapid and sensitive reversed-phase high performance liquid chromatography (HPLC) method was developed for the determination of N-(-4-Chlorophenyl)-6-hydroxy-7-methoxy-2-chromanecarboxamide (KAL-1120), a novel anti-inflammation agent, in the rat plasma. The method was applied to analyze the compound in the biological fluids such as bile, urine and tissue homogenates. After liquid-liquid extraction, the compound was analyzed on an HPLC system with ultraviolet detection at 275 nm. HPLC was carried out using reversed-phase isocratic elution with a $C_{18}$ column, a mobile phase of a mixture of acetonitril (40 v/v%) at a flow rate of 1.0 mL/min. The chromatograms showed good resolution and sensitivity and no interference of plasma. The calibration curve for the drug in plasma was linear over the concentration range of 0.05-50 ${\mu}g$/mL. The intra- and inter-day assay accuracies of this method ranged from 0.06% to 9.33% of normal values and the precision did not exceed 6.28% of relative standard deviation. The plasma concentration of KAL-1120 decreased to below the quantifiable limit at 1.5 hr after the i.v. bolus administration of 2-10 mg/kg to rats ($t_{1/2,({\alpha})}$ and $t_{1/2,({\beta})$ of 2.15 and 26.7 min at a dose of 2 mg/kg, 3.91 and 33.0 min at a dose of 10 mg/kg, respectively). The steady-state volume of distribution ($V_{dss}$) and the total body clearance ($CL_t$) were not significantly altered in rats given doses from 2 to 10 mg/kg. Of the various tissues tested, KAL-1120 was mainly distributed in the lung and heart after i.v. bolus administration. KAL-1120 was detected in the bile by 30 min after its i.v. bolus administration. However, the concentration in the urine after i.v. bolus administration became too low to measure, suggesting that KAL-1120 is mostly excreted in the bile. In conclusion, this analytical method was suitable for the preclinical pharmacokinetic studies of KAL-1120 in rats.

A Study on the Use of Human Antibacterial Peptide LL-37-derived FK-13 as a Cosmetic Preservative (인간 항균펩타이드인 LL-37 유래의 FK-13의 화장품보존제로 활용에 대한 연구)

  • Yun, Hyo-Suk;Choe, Yong-Joon;Yang, Jae-Chan;Min, Hye-Jung
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.6
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    • pp.1568-1576
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    • 2021
  • Here, we conducted the study on the possibility of using FK-13, a short analog of human-derived antibacterial peptide LL-37, as a cosmetic preservative to discover a natural cosmetic preservative that is safe for human body. For the purpose, FK-13 composed of 13 amino acids was synthesized by solid-phase peptide synthesis, and purified using reversed phase-high performance liquid chromatography (RP-HPLC). The purity and molecular weight were confirmed by liquid chromatography-mass spectrometry (LC-MS) analysis. FK-13 showed high antimicrobial activity on the three gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis, and Staphylococcus epidermidis), the three gram-negative bacteria (Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa), and also even the fungus Candida glabrata. FK-13 had a broad spectrum of antibacterial activity, showing a suitability as a cosmetic preservative. In addition, FK-13 showed high thermostability and higher antibacterial activity in a comparative test with existing natural herbal cosmetic and chemical preservatives. Therefore, as FK-13 is a safe material and has high antibacterial activity at a low concentration, it is likely to be applied as a peptide natural cosmetic preservative that can replace existing chemical preservatives.

Theoretical Analysis of Chromatographic Peak Asymmetry and Sharpness by the Moment Method Using Two Peptides

  • Choi Du Young;Row Kyung Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.495-499
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    • 2004
  • The analyses of peak shapes in chromatography are useful in operating chroma­tographic system. The asymmetry and sharpness of a chromatographic peak are estimated by the reversed-phase adsorption of two standard peptides (angiotensin II bradykinin) on $C_{18}$ In this work, the average particle diameters of $C_{18}$ were 5 and 15 $\mu$m, while the pore sizes were 100 and 300 A. The composition of the mobile phase was $50/50\;vol.\;{\%}$ of a binary mixture of acetonitrile and water with $0.1\%$ TFA, and the particles were packed in a stainless column ($4.6{\times}150$ mm). The third and the fourth central movement were calculated from the chromatographic elution curves by moment analysis. The peak asymmetry was determined by two theoretical calculations: the asymmetry factor by elution peak analysis and skewness with moment analysis. The sharpness was estimated by the fourth central moment. In this work, the most acceptable skewness was calculated when the pore size was 300 A. The large excess was observed on small pore size.

Determination of blood concentration of higenamine by high pressure liquid chromatography

  • Park, Sun-Oak;Hong, Chang-Yee;Paik, Seung-Whan;YunChoi, Hye-Sook
    • Archives of Pharmacal Research
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    • v.10 no.1
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    • pp.60-66
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    • 1987
  • A procedure utilizing high pressure liquid chroatography coupled with UV detection is described for the determination of blood concentration of higenamine. Deproteinized serum was pretreated with$C_{18}$(Sep-pak $C_{18}$ cartridge) and the 70% EtOH eluent was applied onto a reversed-phase column ($\mu$ Bondapak $C_{18}]$) with a 15% acetonitrile in 0.05 N $NaH_2$$PO_4$-trichloroacetic acid mixed buffer (pH 2.8) as a mobile phase. With the UV detection at 232 nm, the retention times of higenamine and 1, 2, 3, 4-tetrahydropapaveroline, an internal standard, were 5.2 min and 3.9 min respectively. The blood concentration of higenamine was meausred at regular intervals after i. v. injection of higenamine to rabbit. A drastic decrease in higenamine concentration to 30% of the maximum value obtained immediately after the injection, was observed during the first 1-2 min period and thereafter the rate of decrease was slowed down. The analytical result seemed to coincide with the pharmacological effect of higenamine exerting the maximum chronotropic and hypotensive effect at the completion of the injections which were progressively recovered.

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Antitumor Effects of Fucoidan on Human Colon Cancer Cells via Activation of Akt Signaling

  • Han, Yong-Seok;Lee, Jun Hee;Lee, Sang Hun
    • Biomolecules & Therapeutics
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    • v.23 no.3
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    • pp.225-232
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    • 2015
  • We identified a novel Akt signaling mechanism that mediates fucoidan-induced suppression of human colon cancer cell (HT29) proliferation and anticancer effects. Fucoidan treatment significantly inhibited growth, induced G1-phase-associated upregulation of p21WAF1 expression, and suppressed cyclin and cyclin-dependent kinase expression in HT29 colon cancer cells. Additionally, fucoidan treatment activated the Akt signaling pathway, which was inhibited by treatment with an Akt inhibitor. The inhibition of Akt activation reversed the fucoidan-induced decrease in cell proliferation, the induction of G1-phase-associated p21WAF1 expression, and the reduction in cell cycle regulatory protein expression. Intraperitoneal injection of fucoidan reduced tumor volume; this enhanced antitumor efficacy was associated with induction of apoptosis and decreased angiogenesis. These data suggest that the activation of Akt signaling is involved in the growth inhibition of colon cancer cells treated with fucoidan. Thus, fucoidan may serve as a potential therapeutic agent for colon cancer.

The study of Reversed Two Station Method on Korea-Japan Seismic Data to Obtain Lg Attenuation of Korean Peninsula (한반도 Lg파 감쇠상수 도출을 위한 한일 지진자료의 RTSM 적용에 대한 연구)

  • Chung, Tae-Woong
    • Geophysics and Geophysical Exploration
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    • v.11 no.4
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    • pp.310-317
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    • 2008
  • Lg amplitude decrease $Q^{-1}$ is very important because the Lg phase is the largest seismic phase in Korea if hazardous earthquakes occurred at neighbor nations such as Japan and China. Since reliable $Q^{-1}$ from two station method can be obtained by long pair distance of stations or events, we used seismic stations both Korea and Japans recording Korean events. The obtained $Q^{-1}$ values, however, were anomalously high; these misguided values, inconsistent with tectonic activity of Korean Peninsula, seems to be related with high $Q^{-1}$ values of Japan region.

A Modified Single-Phase Transformerless Z-Source Photovoltaic Grid-Connected Inverter

  • Liu, Hongpeng;Liu, Guihua;Ran, Yan;Wang, Gaolin;Wang, Wei;Xu, Dianguo
    • Journal of Power Electronics
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    • v.15 no.5
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    • pp.1217-1226
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    • 2015
  • In a grid-connected photovoltaic (PV) system, the traditional Z-source inverter uses a low frequency transformer to ensure galvanic isolation between the grid and the PV system. In order to combine the advantages of both Z-source inverters and transformerless PV inverters, this paper presents a modified single-phase transformerless Z-source PV grid-connected inverter and a corresponding PWM strategy to eliminate the ground leakage current. By utilizing two reversed-biased diodes, the path for the leakage current is blocked during the shoot-through state. Meanwhile, by turning off an additional switch, the PV array is decoupled from the grid during the freewheeling state. In this paper, the operation principle, PWM strategy and common-mode (CM) characteristic of the modified transformerless Z-source inverter are illustrated. Furthermore, the influence of the junction capacitances of the power switches is analyzed in detail. The total losses of the main electrical components are evaluated and compared. Finally, a theoretical analysis is presented and corroborated by experimental results from a 1-kW laboratory prototype.

Isolation and Purification of Antimicrobial Substance from the Giant Snail, Achatina fulica (식용 달팽이 [Achatina fulica]로부터 항균성 물질의 분리 및 정제)

  • Kim In-Hae;Hyun Jin-Won;Lee Jae-Hwa
    • Environmental Mutagens and Carcinogens
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    • v.26 no.1
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    • pp.20-24
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    • 2006
  • An antimicrobial substance was purified from the giant snail body extract (Achatina fulica) using solid-phase extraction and separation on HPLC reversed-phase chromatography. The primary structure were determined by a combination of an automated amino acid sequence and MALDI-TOF Mass. Its molecular mass was found to be 1392.64 Da. This result was in excellent agrement with the theoretical molecular mass calculated from the amino acid sequence. purified peptide showed antimicrobial activity in vitro against Escherichia coli D31. This result indicate that giant snail whole body was potentially antimicrobial.

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