• Title/Summary/Keyword: chromatography test

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Simultaneous Analysis of three Marker Components in Hwangryunhaedok-tang by HPLC-DAD (황련해독탕 중 3종 생리활성 물질의 HPLC-DAD 동시 정량분석법 확립)

  • Yang, Hye-Jin;Weon, Jin-Bae;Ma, Jin-Yeul;Ma, Choong-Je
    • YAKHAK HOEJI
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    • v.55 no.1
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    • pp.64-68
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    • 2011
  • In this study, a high performance liquid chromatography-diode array detector method was established, for simultaneous determination of three compounds, berberine, palmatine and geniposide in Hwangryunhaedok-tang, To develop and validate method, $C_{18}$ column (5 ${\mu}M$, 4.6 mm${\times}$250 mm) was used with gradient mobile phase, water containing 0.1% trifluoroacetic acid (TFA) and MeOH at the column temperature of $30^{\circ}C$. UV wavelength was set at 230 and 280 nm. Validation of the chromatography method was evaluated by linearity, precision and accuracy test. Calibration curve of standard components showed good linearity ($R^2$ > 0.9999). The limits of detection (LOD) and limits of quantification (LOQ) varied from 0.05 to 0.17 ${\mu}g/ml$ and 0.15 to 0.53 ${\mu}g/ml$, respectively. The relative standard deviations (RSDs) data of intra-day and inter-day test were in less than 2.99% and 1.90%, respectively. The results of the accuracy test were in the range of 98.36 to 102.52% with RSDs values 0.32 to 1.98%. The results of validation indicated that this method was a very accurate and sensitive assay.

Studies on Quality Control of Domestic Leonurus japonicus Houttuyn (국내 재배산 익모초의 품질관리 연구)

  • Han, Min Woo;Park, Jae Sung;Kwak, Yi Seong;Ahn, Hee Jun;Seo, Jeong Bum;Lee, Young Jong;Park, Chae Kyu
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.6
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    • pp.451-457
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    • 2016
  • Background: Leonurine is a the aerial part of Leonurus japonicus Houttuyn, which has been used as a traditional medicines and is registered in the Korean Pharmacopoeia. Methods and Results: In the present study we performed a heavy metals and thin layer chromatography and measured loss on drying, as well as the contents of total ash, acid-insoluble ash, ethanol soluble compounds, and leonurine, using 15 domestically collected L. japonicus samples. The methods were performed according to the 'crude drugs test of the general test, processes and apparatus', published by MFDS, Korea (2014). The purity test (heavy metals) indicated that levels of Pb and Hg were 0.35 - 3.64, and 0.001 ppm, respectively, whereas the levels of As and Cd were undetectable, and stachydrine was identified by thin layer chromatography ($R_f$ : 0.15). We found that 5.93 - 10.62% (average: $8.58{\pm}1.8%$) of the sample mass was lost during drying, and the contents of total ash, acid-insoluble ash, ethanol soluble compounds, and leonurine were 7.87 - 10.84% (average: $9.62{\pm}0.82%$), 0.99 - 1.76% (average: $1.38{\pm}0.24%$), 16.70 - 23.11% (average: $19.49{\pm}2.14%$) and 0.04 - 0.17% (average: $0.11{\pm}0.04%$) respectively. In addition, HPLC profiling detected leonurine (5.94 min), rutin (16.43 min) and myricetin (26.78 min). Conclusions: We hope that this the rusult of the present study will contribute to the standardization and quality control of Korean herbal medicines.

Spot Test for Amins Acids with Alloxan (Alloxan 에 의한 Amino Acids 의 Spot Test)

  • Kim, Tae-Bong;Hahn, Bo-Sup
    • Journal of the Korean Chemical Society
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    • v.8 no.2
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    • pp.85-87
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    • 1964
  • In order to stabilze alloxan as a reagent for detection of amino acids by spot test, sugars and other reductants were added to the aqueous alloxan solution. It was found that lactose was the best for the purpose. The alloxan reagent containing lactose did not give color change on blank test and was very stable that there was no color change even it was allowed to stand in room temperature for several months. The color reaction with amino acids and some amines was not affected by lactose. This spot test for amino acids is in sensitivity as comparable to that of the previously reported methods and gave color reaction with proline and hydroxyproline to 1${\gamma}$ and 5${\gamma}$ respectively.

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Purification and Characterization of Aminoglycoside-Resistant inhibitior from methylotrophic Actinomycetes (Methanol 자화 방선균으로부터 Aminoglycoside 내성 저해물질의 정제 및 특성)

  • 김현수;신재욱
    • Microbiology and Biotechnology Letters
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    • v.27 no.3
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    • pp.215-222
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    • 1999
  • Methylotrophic actinomycetes No. 155 produced an aminoglycoside antibiotics(AG)-resistant inhibitor. We have previously reported that the inhibitor shows strong inhibition to sisomicin-resistant strain. In order to understand the functions of inhibitor and sisomicin-resistance, characterizations and purification of inhibitor were investigated. Strain No. 155 was tentatively identified as Nocardiopsis sp. based on morphological and some physiological characteristics. In the antimicrobial activity test, the addition of inhibitor to sisomicin showed a reduction effect of MIC on the test strains such as Gram(+), Gram(-) bacteria and yeasts. The combination of the inhibitor and various antibiotics revealed synergistic against E. coli K-12 and B. subtilis PCI 219. The induced intracellular proteins from sisomicin-resistant strain exhibited the sisomicin inactivation by invitro test. And the induced intracellular proteins were inactivated by addition of the inhibitor. The inhibitor compound was purified by anion exchange chromatography(Dowex-1) and HPLC using Asahipak ES-502C column. The purified inhibitor compound was detected in a single peak(above 98.5% purity) through the HPLC analysis.

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A Study on the Methyl Salicylate Dispersion in the Vicinity of Obstacles by Wind Tunnel Test (아음속 풍동을 이용한 구조물 형상 변화에 따른 살리실산메틸 확산 유동 연구)

  • Hong, Chang-Ki;Uhm, Han-Sup;Choi, Seung-Ki;Kim, Youn-Jea
    • The KSFM Journal of Fluid Machinery
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    • v.17 no.6
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    • pp.69-73
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    • 2014
  • To predict flow fields and chemical agent dispersion in urban area, wind tunnel experiments was performed. The agent was adopted MS (methyl salicylate) because the real chemical agent is unsafe. The exact concentration of methyl salicylate was generated by the commercial gas generator (STI-2500) and three different obstacle shapes were applied (i.e., rectangular, cylinder and pyramid). The concentration was measured with the qualified ion mobility sensor and gas chromatography. The data necessary for virtual test method of the real chemical agent were obtained.

Synthesis and Thermal Stability of Polyol Esters (폴리올 에스테르의 합성 및 열 안정성)

  • Chung, Keun-Wo;Baek, Jin-Wook;Kim, Young-Wun;Seo, In-Ok;Han, Jeong-Sik
    • Tribology and Lubricants
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    • v.18 no.5
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    • pp.319-323
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    • 2002
  • Polyol esters were synthesized by condensation reaction of polyols (PE and DiPE) and linear acids such as valeric acid, caproic acid, heptylic acid and caprylic acid. The structures of polyol esters were confirmed by gas chromatography. Hot tube test was used test was used to measure thermal stability of polyol esters and its thermal properties depended on the structure of acid moiety contained in polyol esters.

Cross-reactivity of Amphetamine Analogues in Various Immunoassays (면역분석법에서 암페타민류의 교차 반응성)

  • 박미정;최화경;최상길;손행자;임미애;정희선
    • YAKHAK HOEJI
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    • v.47 no.5
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    • pp.266-270
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    • 2003
  • We evaluated four commercially available methamphetamine immunoassays for their relative cross-reactivities of amphetamine analogues in human urine: Abbott TDx, Vitalab Selectra and on-site test kits (Accusign MET, SD bioline MET). High cross-reactivities were shown at designer's drugs such as methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA) and methylenedioxyethylamphetamine (MDEA) in all of the tested immunoassays. Methoxyphenamine, fenfluramine and phentermine were positive in TDx and Selectra, but were not positive in on-site test kits. Pseudoephedrine, norpseudoephedrine, ephedrine, norephedrine, MDMA, MDA, fenfluramine and phentermine were detected by gas chromatography/mass spectrometry(GC/MS) in false positive urines. Since the overall specificity of any of the devices was not 100%, we found it is important to confirm any positive screening test result, so we developed simultaneous determination of amphetamine analogues in urines. After alkalinization of the urine samples with 6-N NaOH, the analytes were extracted using ethyl acetate, derivatized with pentafluoropropyl anhydride (PFPA) prior at GC/MS analysis.

Identification of Antioxidative Components from Ethanol Extract of Rhus verniciflua STOKES (옻나무 에탄올 추출물로부터 항산화 활성 물질의 구조동정)

  • Kim, In-Won;Shin, Dong-Hwa;Baek, Nam-In
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1654-1660
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    • 1999
  • The free phenolic acid fraction of the chloroform extract from 75% ethanol extract of Rhus verniciflua STOKES (RCF) showed stronger antioxidative activity than BHT, BHA and ${\delta}-tocopherol$ at the same concentration. RCF components were isolated and identified by silica gel column chromatography, thin layer chromatography, mass spectrometer and $^1H-NMR\;and\;^{13}C-NMR$. The antioxidative activity was confirmed by electron donating activity, Rancimat method and thiobarbituric acid test in liposome system. RCF-11 could be further separated into three fractions. The antioxidative active compounds were purified and identified as gallic acid, butin and butein. The RCF-13 was purified and identified as sulfuretin.

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Evaluation on the Safety of ${\gamma}$-Irradiated Angelica gigas Nakai: Stability of Active Components and Safety in Genotoxicity Test (감마선 조사 당귀(Angelica gigas Nakai)의 유효성분 안정성 및 유전독성학적 안전성 연구)

  • 조성기;유영법
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.2
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    • pp.300-306
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    • 2000
  • In the present studies, we assessed the stability of active components and toxicological safety of irradiated Angelica gigas Nakai(Danggui). In order to confirm the stability of active components in the ${\gamma}$-irradiated roots of Danggui, the quantitative analysis of decursin and decursinol angelate of ${\gamma}$-irradiated sample was carried out by high performance liquid chromatographic (HPLC) methods using reverse phase columns and normal phase columns. From the root of Danggui, decursin and decursinol angelate were isolated by a silica gel column chromatography(toluene : ether (1 : 1), Hexane : EtOAc(15 : 1)). And then the structures were confirmed in the 1H and 13C-NMR analysis. The HPLC chromatograms of decursin and decursinol angelate in ${\gamma}$-irradiated Danggui were similar with those of non-irradiated sample. In the examination of in vitro genotoxicity of the water extract from ${\gamma}$-irradiated Danggui using Salmonella reversion assay(Ames test) and micronucleus test in Chinese hamster ovary (CHO) cells, mutagenicity was not exhibited in the two assays with or without metabolic activation. These resutls suggest that active components in the ${\gamma}$-irradiated Danggui should be stable and that the safety of ${\gamma}$-irradiated Danggui could be revealed in further test in vivo.

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Simultaneous Purification of Enterotoxin A and C by Fast Protein Liquid Chromatography (FPLC에 의한 Staphylococcal Enterotoxin A와 C의 동시분리)

  • Lee, Jung-Hee;Kim, Jong-Bae;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.20 no.6
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    • pp.856-861
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    • 1988
  • A new method developed for simultaneous purification of enterotoxin A and C from Staphylococcus aureus strain L 350/1 consisted of chromatography on carboxymethyl (CM)-cellulose using a buffer of variable pH, gel filtration on Ultro gel, and fast protein liquid chromatography(FPLC) using a buffer of variable pH. The enterotoxin A and C were purified by three steps: batchwise adsorption from culture supernatant on Amberlite CG-50; chromatography on CM-cellulose using a buffer of constant pH and molarity; and gel filtration on Sephadex G-75. The purified enterotoxin appeared homogeneous by gel diffusion and polyacrylamide gel electrophoresis. Upon treatment with CM-cellulose using a elution of variable pH, enterotoxin A and C were so close that they were not separated completely. After elution from gels, the enterotoxins appeared as a single peak at the same position. Gel filtration gave a reaction of complete identity to enterotoxin A and C in Ouchterlony immunodiffusion. In FPLC using a CM-cellulose, enterotoxin A and C were simultaneously separated at pH 8.6 and 6.8. When each fraction was performed to gel immunodiffusion, at peak of enterotoxin A and C were not detected each other. In a method of elution by pH-gradient was to be more efficient as a simultaneous separation method in terms of speed, yields and simplicity. The purified toxin A and C were identical to type A and C reference enterotoxin on both disc electrophoresis and Ouchterlony gel diffusion.

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