• 제목/요약/키워드: Elution chromatography

검색결과 415건 처리시간 0.025초

도축혈액으로부터 면역단백질의 효과적인 분리법 (An Effective Method of Isolating Immunoglobulins from Bovine Plasma Proteins)

  • 최인욱;이현정
    • 한국식품과학회지
    • /
    • 제29권5호
    • /
    • pp.1067-1070
    • /
    • 1997
  • Chelating sepharose fast flow gel에 $Cu^{2+}$을 고정화시키고 이들과 단백질과의 친화력 정도에 의해서 용출용매를 통해 단백질을 분획하는 방식의 IMAC법을 이용하여 소의 혈장단백질로부터 IgG의 분리를 시도하였다. 대부분의 혈장단백질은 1차(0.01 M $Na_2HPO_4$, 0.5 M NaCl, pH 4.0)와 2차 용출용매(0.01 M imidazol)에 의하여 용출되었으며 역상 chromatography를 이용하여 각 분획의 단백질조성을 조사한 결과, 1차 용출용매에 의해서는 주로 albumin이, 그리고 2차 용출용매에 의해서는 IgG와 transferrin 등이 IMAC column으로부터 용출되었다. 2차 용출용매에 의해 얻어진 분획으로부터 분자량 100 kD이상의 단백질을 한외여과장치를 이용하여 농축하였을 때, IgG가 효과적으로 분리 정제 되었다.

  • PDF

Elution profiles of metronidazole from calcium sulfate beads

  • Burasarin Ithisariyanont;Saranya Poapolathep;Amnart Poapolathep;Pareeya Udomkusonsri
    • Journal of Veterinary Science
    • /
    • 제24권6호
    • /
    • pp.74.1-74.15
    • /
    • 2023
  • Background: Antibiotic beads are used to treat local bacterial infections by delivering high drug concentrations to infected tissue. Objectives: This study examined the elution characteristics of metronidazole from metronidazole-calcium sulfate (MCa) and metronidazole-calcium-potassium sulfate (MCaK) beads over 20 days and the antibacterial efficacy of the beads after storage. Methods: The MCa and MCaK beads were prepared by mixing 250 mg of metronidazole and 10 g of calcium sulfate hemihydrate with water and a 3% potassium sulfate solution, respectively. The beads were placed in phosphate-buffered saline for the elution study. The metronidazole eluents were determined using high-performance liquid chromatography. The microstructures were examined by scanning electron microscopy (SEM), and the antimicrobial activity was evaluated by a microbioassay. Results: For the 20-day study, the total amount of metronidazole released was greater in the MCa beads than in the MCaK beads by 6.61 ± 0.48 mg (89.11% ± 3.04%) and 4.65 ± 0.36 mg (73.11% ± 4.38%), respectively. The amounts of eluted drugs from the MCa and MCaK beads were higher than the minimum inhibitory concentration at 0.5 ㎍/mL against anaerobic bacteria at both 20 days and 14 days. SEM showed that calcium crystals on the outer surface had dissolved after elution, and thinner calcium crystals were prominent in the MCaK beads. The MCa and MCaK beads exhibited antibacterial activity after setting, followed by storage at room temperature or 4℃ for 21 days. Conclusions: The MCa beads could release more drug than the MCaK beads, but all eluted metronidazole amounts were effective in controlling bacterial infections. Both metronidazole beads could be stored at ambient temperature or in a refrigerator.

환경 오염물질의 정량법 개발과 거동에 관한 연구 ( I ) : 미셀 역상 액체 크로마토그래피에서 페놀과 벤젠 일치환체들의 용리거동 (A Study on the Development of Analytical Methods and Behaviors of Environmental Pollutants ( I ) : Elution Behavior of Monosubstituted Phenols and Benzenes by Micellar Reversed-Phase Liquid Chromatography)

  • 이대운;방은정;조병연
    • 분석과학
    • /
    • 제6권1호
    • /
    • pp.1-8
    • /
    • 1993
  • 음이온 계면활성제인 sodium dodecyl sulfate(SDS)를 이동상으로 한 미셀 액체 크로마토그래피(micellar liquid chromatography : MLC)에서, 시료로 22종의 페놀과 벤젠 일치환체들을 선택하여 이 시료들의 용리거동을 조사하여 MLC계에서의 소수성 효과를 알아 보았다. MLC에서의 머무름과 미셀 농도간의 상관관계를 통해 시료들이 미셀-물간, 변형된 정지상-물 사이에서 분배될 때의 분배계수를 구하였고, 또 이를 이용하여 물-미셀간의 전이 자유 에너지를 구하였다. 이러한 MLC에서 얻은 소수성 파라미터들은 옥탄올-물계와 연관시켜 본 결과 좋은 상관관계를 보였다. 따라서 이 두 계에서 시료의 소수성이 머무름에 중요한 영향을 미침을 알 수 있었다. 한편, MLC를 이용하면 quantitative structure activity relationships(QSAR)에서의 소수성 정량 연구에 적용이 가능하다. 또한 알킬 치환체들에서 탄소 수의 증가에 따른 머무름 관계를 통하여 소수성의 선택성을 알아 보았고, 유기 변형제로 n-프로판올을 첨가하였을 때에도 마찬가지로 MLC계에서의 소수성 파라미터와 옥탄올-물계 사이에 상관관계가 있음을 알았다.

  • PDF

이온교환 칼럼 크로마토그래피를 이용한 난백에서 Lysozyme의 연속추출 (Continuous Separation of Lysozyme from Egg White by Ion Exchange Column Chromatography)

  • 박성준;김현석;김현위;안태회;박기문;최춘언
    • 한국식품과학회지
    • /
    • 제22권6호
    • /
    • pp.711-715
    • /
    • 1990
  • 식품 및 의약품 원료로 이용되고 있는 Iysozyme을 난백으로부터 이온교환 크로마토그래프를 이용하여 연속추출하였다. 사용된 수지는 지금까지 수율, 순도, 작동용이성에서 가장 효과적이라고 밝혀져 있는 Duolite C-464 양이온교환수지를 선택하여 $Na^+$형으로 $pH\;7.9{\pm}0.1$로 평형화하여 사용하였다. 조제용 자동 Liquid Chromatography(LC) system(column size ; i.d. 50 mm, bed volumn ; 1020 ml)에서 수지 평형화, 난백접촉, rinse, lysozyme용출 순으로 cycle을 연속적으로 반복하면서 시행하고 그 재현성을 관찰하기 위해 각 cycle별 rinse끝점과 용출끝점의 UV level을 비교하였다. 그 결과 rinse끝점은 19 cycle까지 30% 이하를 유지하였다. 용출 끝점은 17 cycle까지는 30% 이하에서 비교적 안정하였으나 18 cycle 이후 부터는 50% 이상으로 용출능력이 현저하게 감소하였다. 또한, 회수율 비교에서 17cycle까지는 90% 이상을 유지하다가 18cycle에서는 72%, 19 cycle에서는 65%로 격감하였다. 본 추출 정제 lysozyme은 전기영동상 단일 band로 나타났고, densitometer로 측정한 순도는 99% 이상이었다.

  • PDF

손바닥선인장의 항염증 활성 (Anti-inflammatory Activity of Opuntia ficus-indica)

  • 박은희;황성은;강자훈
    • 약학회지
    • /
    • 제42권6호
    • /
    • pp.621-626
    • /
    • 1998
  • Cactus (Opuntia ficus-indica var saboten Makino) is a tropical or subtropical plant, which is widely used as folk medicine for burned wound, edema and indigestion. We previously found that the ethanol extract of cactus stem showed anti-inflanunatory action. This investigation was designed to isolate the active fraction of anti-inflanimatory action from cactus stem by solvent extraction and colunm chromatography. Carrageenan-induced paw edema in rats and acetic acid-induced writhing test in mice were used as animal models to search anti-inflammatory and analgesic activities. respectively. The ethanol extract of cactus stem was consecutively extracted with hexane, ethyl acetate, and n-butanol. The hexane fraction was the most effective in carrageenan-induced paw edema, and then was separated in colunm chromatography of silica gel by the elution with hexane/ethyl acetate mixture. The most effective fraction 1 was separated in a second colunm chromatography by eluting with hexane/diethyl ether mixture. The most effective fraction 1-5 was obtained, and separated in a third column chromatography by eluting with hexane/chloroform mixture. It produced the most effective fraction 1-5-1. Moreover, fraction 1-5-1 showed an inhibitory effect on acetic acid-induced writhing in the doses of 30mg/kg and 60mg/kg,p.o.,indicating that it also contained analgesic activity.

  • PDF

Determination of Persistent Organochlorine Pollutants in Rat Hair by Gas Chromatography-Mass Spectrometry

  • Shin, Ho-Sang;Pyo, Hee-Soo;Park, Song-Ja
    • Bulletin of the Korean Chemical Society
    • /
    • 제25권10호
    • /
    • pp.1489-1494
    • /
    • 2004
  • A gas chromatography/mass spectrometric assay method was developed for the determination of persistent organochlorine pollutants (POPs) in hair. For the exact extraction study was used hair of rat exposed with POPs. Sonication of the hair matrix with 3 M HCl solution in methylene chloride of the extraction methods studied was the most efficient and rapid sample preparation method. After sonication of rat hair was achieved clean up with a solid phase extraction procedure using silica gel-florisil. Elution was performed with 8 mL of methylene chloride. The eluate was concentrated to approximately 100 ${\mu}L$ and analyzed by gas chromatography-mass spectrometry (GC-MS). Detection limits of POPs were in the concentration range of 0.6-1.2 ng/g in rat hair. Aldrin, dieldrin, p,p-DDT and mirex were dosed rat for 4 weeks at concentration of 0.01 mg/L in drinking water and detected in rat hair at concentration of 2.8, 11.3, 7.9 and 15.6 ng/g, respectively. Aldrin and p,p-DDT were metabolized to dieldrin and p,p-DDE, which were detected in concentration of 9.7 and 2.9 ng/g in rat hair, respectively. The developed method may be valuable to be used to analyze POPs in human hair.

수수 줄기에 함유(含有)된 타감물질(他感物質)의 분리(分離) 및 특성(特性) 구명(究明) (Isolation and Characterization of Allelopathic Substances from Sorghum Stem)

  • 김상열;에스 케이 디다타;알 피이 로블레스;김길웅;이상철;신동현
    • 한국잡초학회지
    • /
    • 제14권2호
    • /
    • pp.156-162
    • /
    • 1994
  • 수수 줄기에 함유된 타감물질(他感物質)중 가장 강한 활성을 나타내는 물질을 rapid chromatography, flash flow column chromatography로 분리하고 thin layer chromatography와 HPLC로 정제하여 GC-MS로 분석하였다. Butanol, acetic acid 및 water의 용매 조합을 달리하여 타감물질을 분리한 결과 butanol (8) : actic acid (1) : water (1) 분획에서 억제효과가 가장 크게 나타났으며, flash flow column chronatography와 TLC로 분리 한 결과 가장 활성을 나타내는 물질은 Rf 0.71에서 나타났으며 HPLC로 순수분리한 결과 Rt 20.40min에서 elution되었다. 이 물질은 주황색을 띄며 methanol에 용해성이 있었다. 정제된 물질을 GC-MS로 분석한 결과 예상되는 물질은 1-methyl-1-(2-propinonyl)-hydrazine, 1-aziridineethanol, 5-chloro-2-pentanone, 2-(methylseleno)-ethanamine 중한 물질일 것으로 추측되었다.

  • PDF

쑥(Artemisia princeps Pampan) 추출 성분의 암세포증식 억제효과 (Inhibitory Effect of Artemisia princeps Pampan.. Extract on Growth of Cancer Cell Lines)

  • 황윤경
    • Journal of Nutrition and Health
    • /
    • 제31권4호
    • /
    • pp.799-808
    • /
    • 1998
  • To investigate the antitumor activity of mugwort (Artemisia princeps Pampan), petroleum ether extract of mugwork was partially purified by a silica gel chromatography. Among several fractions, the fraction which was obtained under the elution with acetone, showed potent cytotoxicity against mouse leukemia cell line(Ll210), human colon cancer cell line (HCT-48) and human hepatoma cell line (Hep G2) , but was less effective with normal cell line(mouse embryo cell). Acetone fraction appeared to be glycolipid by Benedict test and the major fatty acids of the lipid were C16 ; 0 , C 18: 3by GC/MS analysis.

  • PDF

녹두로부터 렉틴 성분의 분리 정제 (Isolation and Purification of Lectin from Phaseolus radiatus)

  • 정시련;홍승수;전경희
    • 약학회지
    • /
    • 제27권3호
    • /
    • pp.221-227
    • /
    • 1983
  • New lectins, lymphoagglutinating lectins from mung beans (MBLA) have been isolated and purified. Mung beans crude extracts were made with 0.15M NaCl and these were purified through anionic exchange chromatography. Four fractions were obtained from DEAE Sephadex A-50 by salt gradients elution. Lectin activity, enzyme activity, protein assay, identification of purity by polyacrylamide gel electrphoresis and immunochemical studies were carried out with these four fractions. Through these results, it can be suggested that 0.2M fraction is newly found potent MBLA. There were some relationships with MBLA and L-PHA but no similarities were observed between MBLA and E-PHA.

  • PDF