• Title/Summary/Keyword: Reversed phase chromatography

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Separation and Detection of Nonchromophore Aliphatic Compounds by Reversed-Phase Liquid Chromatography using Ultraviolet-Absorbing Reagent (자외선 흡수물질을 이용한 역상 액체 크로마토그라피에 의한 비흡수 지방족 화합물들의 검출과 분리)

  • Lee Seung-Seok;Kang Sam-Woo;Oh Hae-Beom
    • Journal of the Korean Chemical Society
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    • v.35 no.4
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    • pp.397-404
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    • 1991
  • Nonchromophore compounds such as aliphatic acids, alcohols and tetraalkylammonium salts could be detected by indirect photometric detection on the revered-phase liquid chromatography. Benzyltriethylammonium bromide(BTEAB) was used as a detection reagent. Also, the retention mechanism and response of samples were investigated to the several factors such as pH, temperature, and concentration of MeOH as well as concentration of detection reagents in mobile phase. And some mixture of samples were able to be separated under optimum condition.

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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.

Retention Behavior of Organic Compounds on Reversed-Phase Column expected by Van der Waals Volume (Van der Waals Volume을 이용한 역상 컬럼에서의 유기화합물들의 용출거동)

  • Park, Wun-Kyu;Lee, Yong-Moon;Moon, Dong-Cheul;Kang, Jong-Seong
    • Analytical Science and Technology
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    • v.6 no.4
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    • pp.383-390
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    • 1993
  • Retention behaviors of aliphatic and aromatic compounds were investigated with Van der Waals volume which represented the molecular size. The organic solvents, methanol, acetonitrile and tetrahydrofuran, were mixed with water at various ratio, respectively. The selectivity of organic solvents were tested by change of column temperature. The capacity factor was increased linearly according to the enlargement of molecular size. Therefore, Van der Waals volume was useful to predict the elution of organic molecules in reversed-phase column. The order of elution capacity of solvents was methanol

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Influence of pH, Temperature, Ionic Strength and Metal Ions on the Degradation of an Iridoid Glucoside, Aucubin, in Buffered Aqueous Solutions (완충 수용액중 pH, 온도, 이온강도 및 금속이온이 Aucubin의 분해에 미치는 영향)

  • Chun, In-Koo;Cho, Young-Mee
    • Journal of Pharmaceutical Investigation
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    • v.25 no.3
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    • pp.239-247
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    • 1995
  • The physico-chemical stability of aucubin, a hepatoprotective iridoid glucoside, in buffered aqueous solutions was studied using a stability-indicating reversed-phase high performance liquid chromatography. The degradation of aucubin followed the pseudo-first-order kinetics. In strong acidic regions, aucubin was rapidly degraded by the specific acid catalysis, forming dark brown precipitates. From the rate-pH profiles, it was found that aucubin was most stable at the pH of about 10. From the temperature dependence of degradation, activation energies for aucubin at pH 2.1 and 4.9 were calculated to be 22.0 and 24.3 kcal/mole, respectively. The shelf-life $(t_{90%})$ for aucubin at pH 9.07 and $20^{\circ}C$ was predicted to be about 603 days. A higher ionic strength accelerated the degradation of aucubin at pH 4.01. The effect of metal ions on the degradation rate of aucubin at pH 7.16 was in the rank order of $Cu^{2+}\;>\;Fe^{3+}\;>\;Co^{2+}\;>\;Fe^{2+}\;>\;Mg^{2+}$. On the other hand, $Mn^{2+}\;and\;Ba^{2+}$ slowed the degradation rate.

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Determination of Rosmarinic Acid and Caffeic Acid from Perilla frutescens var. japonica and var. acuta by Reversed-Phase HPLC (RP-HPLC를 이용한 백소엽(白蘇葉)과 자소엽(紫蘇葉)의 카페익산과 로즈마린산 분석)

  • Kim, Byung-Youn;Jeong, Ji-Seon;Kwon, Ha-Jeong;Lee, Je-Hyun;Hong, Seon-Pyo
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.67-72
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    • 2008
  • Objectives : This study presents a high performance liquid chromatography methods for the quantitative and qualitative analysis of rosmarinic acid (RA) and caffeic acid (CA) in Perilla frutescens var. japonica and var. acuta. Methods : Chromatographic separation was performed using a mixture of methanol, water and formic acid (35 : 64.2 : 0.8) with a reversed-phase column (Gemini C18, 4.6 ${\times}$ 150 mm, 3 ${\mu}m$). The analyses were detected at UV (280 nm). Results : The samples were extracted with 50% EtOH under reflux for 1 h, and simultaneous determination for RA and CA in hyang-so-san and haeng-so-san was possible without interference peaks Conclusions : According the results, we developed a determination method for RA and CA in Perillae Folium. Owing to Perilla frutescens var. japonica and var. acuta did not show significant difference in contents of RA and CA, both Perilla frutescens could be available as herbal medicine.

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Comparison of 2-D RP-RP MS/MS with 1-D RP MS/MS for Proteomic Analysis (단백체 분석을 위한 일차원 및 이차원 역상크로마토그래피의 비교)

  • Moon, Pyong-Gon;Cho, Young-Eun;Baek, Moon-Chang
    • YAKHAK HOEJI
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    • v.54 no.5
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    • pp.377-386
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    • 2010
  • Single-dimensional (1-D) and two-dimensional (2-D) LC methods were utilized to separate peptides from various sources followed by MS/MS analysis. Two-dimensional ultra-high performance liquid chromatography is a useful tool for proteome analysis, providing a greater peak capacity than 1-D LC. The most popular 2-D LC approach used today for proteomic research combines strong cation exchange and reversed-phase LC. We have evaluated an alternative mode for 2-D LC of peptides using 2-D RP-RP nano UPLC Q-TOF Mass Spectrometry, employing reversed-phase columns in both separation dimensions. As control experiments, we identified 129 proteins in 1-D LC and 322 proteins in 2-D LC from E. coli extract peptides. Furthermore, we applied this method to rat primary hepatocyte and a total of 170 proteins were identified from 1-D LC, and 527 proteins were identified from all 2-D LC system. The in-depth protein profiling established by this 2-D LC MS/MS from rat primary hepatocyte could be a very useful reference for future applications in regards to drug induced liver toxicity.

Isolation of Iron and Calcium-Binding Peptides from Cottonseed Meal Protein Hydrolysates (면실박 단백질로부터 가수분해물 제조 및 철분, 칼슘 결합 펩타이드의 분리)

  • Choi, Dong-Won;Kim, Nam-Ho;Song, Kyung Bin
    • Journal of Applied Biological Chemistry
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    • v.55 no.4
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    • pp.263-266
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    • 2012
  • Isolation of iron and calcium-binding peptides derived from cottonseed meal protein (CMP) hydrolysates was investigated. The degree of hydrolysis of CMP by Flavourzyme was monitored using trinitrobenzenesulfonic acid method and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Enzymatic hydrolysis of CMP for 12 h was sufficient for the preparation of CMP hydrolysates, and the hydrolysates were membrane-filtered under 3 kDa as a molecular weight. The filtered solution was fractionated using Q-Sepharose fast flow, Sephadex G-15, and reversed phase-high performance liquid chromatography for iron and calcium-binding peptides. As a result, F51 fraction was obtained as the best candidate for calcium and iron chelation, and the isolated iron and calcium-binding peptides can be used as functional food additives, similar to iron and calcium supplements.

Simultaneous Determination of Chlorogenic Acid and Linarin in Chrysanthemum Sibiricum Fisher by Reversed-Phase High Performance Liquid Chromatography (역상 액체 크로마토그래피에 의한 구절초 중 Chlorogenic Acids와 Linarin의 동시 정량분석)

  • Kim, Taek-Jae;Lee, Tae-Ryong;Park, Ho-Koon
    • Journal of the Korean Chemical Society
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    • v.35 no.6
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    • pp.720-724
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    • 1991
  • Simultaneous determination of chloragenic acid (CA), 3,4-o-dicafeoyl quinic acid(3,4-DCQA), 4,5-o-dicaffeoyl quinic acid (4,5-DCQA) and linarin in Chrysanthemum sibiricum Fisher was newly established by a reversed-phase high performance liquid chromatography (HPLC). Sample was extracted with 20 ml methanol for 4 hrs. The extract was cleaned up by using Sep-Pak $C_18$ cartridge and 4 ml methanol-$H_2$O(1 : 1) as eluent. Their determination was performed by means of RP-HPLC with Bondapak $C_18$ column (30 cm ${\times}$ 3.9 mm i.d., 10 ${\mu}m$ and gradient elution mode as methanol-5 mM $H_3PO_4$ solution (30 : 70). The established method was applied to various samples purchased. As a result, their content ranges showed to be 0.35~0.55% for CA, 0.46~0.76% for 3,5-DCQA, 0.077~0.23% for 4,5-DCQA and 0.16~2.72% for linarin, respectively.

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Determination of Aloesin in Plasma by High-Performance Liquid Chromatography as Fluorescent 9-Anthroyl Derivative

  • Kim, Kyeong-Ho;Lee, Jin-Gyun;Park, Jeong-Hill;Shin, Young-Geun;Lee, Seung-Ki;Cho, Tae-Hyung
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.651-656
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    • 1998
  • A sensitive high-performance liquid chromatographic (HPLC) method for the determination of aloesin in plasma was developed. After solid-phase extraction from plasma and derivatization of aloesin and compound AD-1, which was prepared from aloesin as a internal standard, with 9-anthroylnitrile in the presence of quinuclidine, the derivatives were separated on a Inertsil ODS-3 column using acetonitrile/methanol/water (3:1:6) as a mobile phase, and detected fluorimetrically at 460nm with excitation at 360nm. The detection limit of aloesin was 3.2ng/ml in plasma (S/N=3).

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Resolution of Salbutamol Enantiomers in Human Urine by Reversed-phase High Performance Liquid Chromatography after Derivatization with (S)-(-)-${\alpha}$-methylbenzyl isocyanate

  • Kim, Kyeong-Ho;Kim, Tae-Kyun;Kwon, Young-Hee;Sohn, Young-Teak
    • Archives of Pharmacal Research
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    • v.20 no.5
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    • pp.486-490
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    • 1997
  • A stereospecific HPLC method has been developed for the resolution of the enantiomers of salbutamol in human urine. After solid-phase extraction and derivatization with (S)-(-)-${\alpha}$-methyl-benzyl isocyanate, the diastereomeric derivatives were resolved $(R_s=1.59)$ on $5{\mu}M$ octadecylsilan column using 47% methanol as a mobile phase with fluorescence detection. The detection limit of each enantiomer was 10 ng/ml (S/N=3).

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