• Title/Summary/Keyword: reversed phase

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Sensitive Determination of Pinaverium Bromide in Human Plasma by LC-ESI-MS/MS : Applicability to Oral Bioavailability Determination (LC-ESI-MS/MS를 이용한 생체시료 중 브롬화피나베리움의 고감도 분석 및 이를 이용한 생체이용률 평가)

  • Park, Seok;Lee, Ye-Rie;Kim, Ho-Hyun;Lee, Hee-Joo;Kim, Yoon-Gyoon;Youm, Jeong-Rok;Han, Sang-Beom
    • Journal of Pharmaceutical Investigation
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    • v.34 no.6
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    • pp.513-519
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    • 2004
  • A sensitive method for quantification of pinaverium bromide in human plasma was established using liquid chromatography-electrospray ionization tandem mass spectrometry(LC-ESI-MS/MS). Glimepiride was used as internal standard. Pinaverium bromide and internal standard in plasma sample were extracted using tert-butylmethylether(TBME). A centrifuged upper layer was then evaporated and reconstituted with mobile phase of acetonitrile-5 mM ammonium formate (80/20, pH 3.0). The reconstituted samples were injected into a $C_{18}$ reversed-phase column. Using MS/MS with multiple reaction monitoring (MRM) mode, pinaverium and glimepirde were detected without severe interference from human plasma matrix. Pinaverium produced a protonated precursor ion $([M+H]^+)$ at m/z 510.3 and a corresponding product ion at m/z 228.9. Internal standard produced a protonated precursor ion $([M+H]^+)$ at m/z 491.5 and a corresponding product ion at m/z 352.0. Detection of pinaverium bromide in human plasma was accurate and precise, with limit of quantitation at 0.5 ng/ml. The method has been successfully applied to bioavailability study of pinaverium bromide tablet in Korean healthy male volunteers. Pharmacokinetic parameters such as $AUC_t,\;C_{max},\;T_{max},\;K_{el}\;and\;t_{1/2}$ were calculated.

HPLC Analysis and Extraction Methods of Decursin and Decursinol Angelate in Angelica gigas Roots (당귀(Angelica gigas) 중 Decursin 및 Decursinol Angelate 추출 방법과 HPLC 분석)

  • Lee, Jang-Hoon;Chae, Hee-Jeong;Kim, Dong-Hyun;Lee, Sang-Hyun;Park, Sang-Yong;Kang, Young-Goo
    • Korean Journal of Pharmacognosy
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    • v.34 no.3 s.134
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    • pp.201-205
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    • 2003
  • This paper is intended as an investigation of the analysis of high-performance liquid chromatography and the method of extraction of decursin and decursinol angelate in Angelica gigas roots. There are three kinds of extraction methods: distilled water, 50% EtOH and 100% EtOH. The condition of HPLC was obtained on a reversed-phase column $(Polarity\;dC_{18},\;4.6{\times}250 mm,\;5\;{\mu}m)$ using a phosphate buffer-acetonitrile-sodium lauryl sulfate as the mobile phase. Under these chromatographic conditions, UV detector was 230 nm, column temperature $30^{\circ}C$ and the speed of a current 1.0 ml/min, respectively. The results of extraction with distilled water, 50% EtOH and 100% EtOH in Angelica gigas roots were as follows. The concentrations of decursin and decursinol angelate were 182 and 153 ppm (distilled water), 3,142 and 2,547 ppm (50% EtOH) and 3,341 and 2,778 ppm (100% EtOH). There were high positive correlations between the concentrations of decursin and EtOH (r=0.8928, p<0.01) and decursinol angelate and EtOH (r=0.9009, p<0.01).

Screening and Purification of a Novel Antibacterial Peptide, cgCAFLP, Against Skin Pathogens from the Extract of the Pacific Oyster Crassostrea gigas from Buan in Korea (부안산 참굴(Crassostrea gigas) 추출물로부터 피부 상재균에 대한 새로운 항균 펩타이드, cgCAFLP의 탐색 및 정제)

  • Lee, Ji-Eun;Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.6
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    • pp.927-937
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    • 2021
  • This study was performed to screen the antimicrobial activities of the extract from the Pacific oyster Crassostrea gigas against skin pathogens and to purify the relevant antibacterial peptide. The acidified extract showed potent antibacterial activities against gram-positive and gram-negative bacteria but showed no activity against Candida albicans and no significant cell toxicity. Among acne-causing pathogens, the acidified extract showed potent antibacterial activity only against Staphylococcus aureus, and its antibacterial activity was completely abolished by treatment with trypsin or chymotrypsin, and was inhibited by salt treatment. The acidified extract showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. Based on antimicrobial activity screening and cytotoxic effects, a novel antibacterial peptide was purified from the acidified gill extract using solid-phase extraction, cation-exchange, and reversed-phase HPLC. The resulting peptide had a molecular weight of 4800.8 Da and showed partial sequence homology with the carbonic anhydrase 4 (CA4) protein in the hard-shelled mussel. Overall, we purified a novel antibacterial peptide, named cgCAFLP, which is related to carbonic anhydrase 4 (CA4) protein, against skin pathogens. Our results suggest that the Pacific oyster extract could be used as an additive to control some acne-related skin pathogens (S. aureus).

Determination of diallyl disulfide in garlic by reversed-phase high performance liquid chromatography (역상 고성능 액체 크로마토그래피를 이용한 마늘에서 diallyl disulfide의 분석)

  • Wan, Xiaolong;Polyakova, Yulia;Row, Kyung Ho
    • Analytical Science and Technology
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    • v.20 no.5
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    • pp.442-447
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    • 2007
  • The properties of garlic (Allium sativum L.) are attributed to organosulfur compounds. In this paper, an analytical technique with a rapid and simple sample preparation procedures for determination of diallyl disulfide (DADS) in garlic was reported. The DADS was simply extracted with various solvents (methanol, benzene or tetrahydrofuran) from garlic and prepared for HPLC analysis. From the results, the methanol was select as an optimal extraction solvent. The mobile phase was composed from methanol and water, and the gradient elution mode was applied. 0.61 mg of DADS per g garlic powder can be extracted with methanol. This work offers some advantages over the currently accepted techniques and would be useful for chemical and biological studies of garlic and its products.

1,1-Diphenyl-2-picrylhydrazyl Radical Scavenging Compounds of Fraxini Cortex

  • Kim, Hyun-Chul;An, Ren-Bo;Jeong, Gil-Saeng;Oh, Seung-Hwan;Kim, Youn-Chul
    • Natural Product Sciences
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    • v.11 no.3
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    • pp.150-154
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    • 2005
  • The radical scavenging effect of the MeOH extract of Fraxini Cortex on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical was examined. The $CH_2Cl_2$-and n-BuOH-soluble fractions of MeOH extract showed the promising DPPH radical scavenging effects, and further purified by silica gel, Sephadex LH-20 column chromatography, and reversed-phase C-18 MPLC to yield five coumarins, esculetin (1), fraxidin (2), fraxetin (3), fraxidin $8-O-{\beta}-D-glucopyranoside$ (fraxin methyl ether) (5), esculin (6), and a secoiridoid oleuropein (4), and a coumarin-secoiridoid escuside (7). Compounds 1, 3, and 4 showed potent DPPH radical scavenging effects, exhibiting $IC_{50}$ values of 14.68, 9.64, and $22.03\;{\mu}M$, respectively. Compounds 6 and 7 also showed moderate effects with $IC_{50}$ values of 147.79 and $72.73\;{\mu}M$, respectively. L-Ascorbic acid was used as a positive control and exhibited the $IC_{50}$ value of $50.31\;{\mu}M$.

Separation of Cations on the Hydrophobic Reagent Coated Column (Hydrophobic Reagent를 흡착시킨 분리관에서의 양이온 분리)

  • Tae Yoon Eom;Kih Soo Cho;Youn Doo Kim
    • Journal of the Korean Chemical Society
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    • v.31 no.4
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    • pp.328-333
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    • 1987
  • 1-eicosylsulfate($C_{20}H_{41}SO_4^-$) was coated onto 5 ${\mu}m$ C18 reversed phase column. Adsorption equilibrium was reached when 1.2l of $1.46{\times}10^{-4}M$ in 25% acetonitrile solution passed through the column. This coated column proved to be stable up to 10l of 0.05 M ${\alpha}$-HiBA eluent used. The column efficiencies of Eu, Nd, Ce and La measured were about 0.04 mm of HETP using 0.15M ${\alpha}$-HiBA eluent at a flow rate of 1.0ml/min. This column was used to separate some lanthanides and transition metal ions, andmshowed an excellent resolution, respectively.

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A New Flavonol Glycoside from Tristemma hirtum (Melastomataceae)

  • Kenfack, Joseph Nandjou;Ponou, Beaudelaire Kemvoufo;Kuhlborn, Jonas;Teponno, Remy Bertrand;Nono, Raymond Ngansop;Fouedjou, Romuald Tematio;Opatz, Till;Park, Hee Juhn;Tapondjou, Leon Azefack
    • Natural Product Sciences
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    • v.24 no.3
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    • pp.213-218
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    • 2018
  • Chemical investigation of the plant Tristemma hirtum P. Beauv (Melastomataceae) resulted to the isolation of a new flavonol glycoside named quercetin-7-O-${\alpha}$-D-arabinofuranoside (1), together with nine known compounds including 3'-hexadecanoyl-2'-(9aZ)-tetradecanoyl-glycerol 1'-O-[${\beta}$-D-galactopyranosyl-(1'' ${\rightarrow}$ 6'')-${\alpha}$-D-galactopyranoside] (2), arjunolic acid (3), ${\beta}$-sitosterol-3-O-${\beta}$-D-glucopyranoside (4), terminolic acid (5), quercetin (6), asiatic acid (7), maslinic acid (8), $1{\beta}$-O-galloylpedunculagin (9) and 6-hydroxyapigenin 7-O-${\beta}$-D-glucopyranoside (10) from the methanol extract using normal and reversed phase column chromatography. The structures of these compounds were determined by comprehensive interpretation of their spectral data mainly including 1D- 2D-NMR ($^1H-^1H$ COSY, HSQC, and HMBC) spectroscopic and ESI-TOF-MS mass spectrometric analysis.

Determination of Nucleobases and Nucleosides in Panax and Acanthopanax genus (인삼속(人蔘屬)과 오가피속(五加皮屬)의 Nucleobases 및 Nucleosides의 분석(分析))

  • Cho, Yung Hyun;Hong, Soon Keun;Park, Nae Jung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.1
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    • pp.1-6
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    • 1983
  • Nucleobases and nucleosides in Panax and Acanthopanax genus were determined by high-performance liquid chromatography. Chromatography was performed on a reversed-phase system with ${\mu}$ Bondapak $C_{18}$ column using phosphate buffer and 80% methanol gradient. Content of each nucleobase in two genera was about 0-2mg/100g. Panax was contained guanosine and/or adenosine ca. 15-22mg/100g;and Acanthopanax guanosine ca. 3-8mg/100g and adenosine ca. 2-7mg/100g. Considerable amounts of cytidine, uridine, inosine, and thymidine were also detected in two genera.

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Molecular Identification of Four Different α-amylase Inhibitors from Baru (Dipteryx alata) Seeds with Activity Toward Insect Enzymes

  • Bonavides, Krishna B.;Pelegrini, Patricia B.;Laumann, Raul A.;Grossi-De-Sa, Maria F.;Bloch, Carlos Jr.;Melo, Jorge A.T.;Quirino, Betania F.;Noronha, Eliane F.;Franco, Octavio L.
    • BMB Reports
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    • v.40 no.4
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    • pp.494-500
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    • 2007
  • The endophytic bruchid pest Callosobruchus maculatus causes severe damage to storage cowpea seeds, leading to economical losses. For this reason the use of $\alpha$-amylase inhibitors to interfere with the pest digestion process has been an interesting alternative to control bruchids. With this aim, $\alpha$-amylase inhibitors from baru seeds (Dipteryx alata) were isolated by affinity chromatographic procedures, causing enhanced inhibition of C. maculatus and Anthonomus grandis $\alpha$-amylases. To attempt further purification, this fraction was applied onto a reversed-phase HPLC column, generating four peaks with remarkable inhibition toward C. maculatus $\alpha$-amylases. SDS-PAGE and MALDI-ToF analysis identified major proteins of approximately 5.0, 11.0, 20.0 and 55 kDa that showed $\alpha$-amylase inhibition. Results of in vivo bioassays using artificial seeds containing 1.0% (w/w) of baru crude extract revealed 40% cowpea weevil larvae mortality. These results provide evidence that several $\alpha$-amylase inhibitors classes, with biotechnological potential, can be isolated from a single plant species.

A Study on the Extraction Efficiency of Berberine in Coptidis Rhizoma (황련 중 berberine의 추출효율에 관한 연구)

  • Rhee, Jae Seong;Park, Ho Koon;Woo, Eun Ran;Lee, Eun Ju;Ahn, Dug Kyun;Youn, Won Sik;Shin, Kwang Ho
    • Analytical Science and Technology
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    • v.10 no.2
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    • pp.83-90
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    • 1997
  • In this paper, optimum extraction condition for berberine which is the indan companent of Coptidis rhizoma was examined at various conditions in the aspect of temperature, solvent and time followed by quantitation with reversed phase HPLC. The efficiency for extraction has been shown the best at $60^{\circ}C$ with 2~3 hours reflux time in methanol or acetic acid. Any significant effect can not be shown from the addition of SLS to extraction solvent.

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