• 제목/요약/키워드: HPLC Chromatography

검색결과 2,212건 처리시간 0.028초

Ginsenoside의 새로운 분리.정제 방법 (New Efficient Method for Isolation and Purification of Ginsenosides)

  • 김세원;황석연
    • Journal of Ginseng Research
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    • 제22권4호
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    • pp.284-288
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    • 1998
  • This study was carried out to establish a new efficient method for isolation and purification of ginsenosides. Silica gel column chromatography, having been used for the isolation of ginsenosides, is advantageous to obtain a large amount of ginsenosides. However, it has a disadvantage to isolate ginsenosides to their highest purity. In addition, normal-or reverse-phase HPLC method thus far reported is confined to quantitative analysis. Especially, it has not been possible to isolate racemic 20(S)- and 20(R)-ginsenoside Rg2. In this experiment, isolation and purification of ginsenosides were accomplished by Diaion HP-20 adsorption chromatography, silica gel column chromatography, recrystalization and Prep. HPLC with or without Prep. TLC. From this study, we could establish a new efficient method for isolation and purification of 9 major and/or minor ginsenosides.

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셀레늄이 강화된 이스트에서 셀레늄 단백질의 분리 및 확인 (Separation and identification of selenoproteins in selenium-enriched yeast)

  • 김경미;박용남
    • 분석과학
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    • 제26권6호
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    • pp.357-363
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    • 2013
  • Selenium-containing proteins were separated from selenium-enriched yeast (SEY) using Trizol$^{(R)}$ reagent followed by anion exchange (AE) chromatography. This method is simpler and less time consuming than electrophoresis. Five selenium containing proteins were identified by on-line AE HPLC-ICP/MS (high performance liquid chromatography-inductively coupled plasma/mass spectrometry). Each protein was enzymatically hydrolyzed to seleno-amino acids and separated with RP (reverse phase) HPLC for the identification of selenoproteins.

한국산 호박의 Carotenoid 색소 (Carotenoid pigment of Pumpukin Culitivated in Korea)

  • 박용곤
    • 한국식품영양학회지
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    • 제12권5호
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    • pp.508-512
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    • 1999
  • In order to qualify and quantify carotenoid pigment of pumpukin were analyzed by HPLC column chromatography and spectrophotometry. Eightyseven percent of total carotenoids in the pumpkin was in the fiber amounting 65.33mg%. however flesh and skin contained 6.61 and 3.31mg% respectvely. A silica gel column chromatography of the carotenoids produced 2 different peaks. The components eluted in peak II showed blue color when treated with hydrochloric acid. The fact suggested it contained epoxide and the component in peak II was a monol. By the separation with HPLC. $\alpha$ and $\beta$-carotene were the major compounds in the carotenoids. In fiber the total $\beta$-carotene content was 1.6 times higher than in the flesh and skin. The fiber contained less unidentified compounds than other part which contained 27.3~32.8%.

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Microcystin Detection Characteristics of Fluorescence Immunochromatography and High Performance Liquid Chromatography

  • Pyo, Dong-Jin;Park, Geun-Young;Choi, Jong-Chon;Oh, Chang-Suk
    • Bulletin of the Korean Chemical Society
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    • 제26권2호
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    • pp.268-272
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    • 2005
  • Different detection characteristics of fluorescence immunochromatography method and high performance liquid chromatography (HPLC) method for the analysis of cyanobacterial toxins were studied. In particular, low and high limits of detection, detection time and reproducibility and detectable microcystin species were compared when fluorescence immunochromatography method and high performance liquid chromatography method were applied for the detection of microcystin (MC), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa. A Fluorescence immunochromatography assay system has the unique advantages of short detection time and low detection limit, and high performance liquid chromatography detection method has the strong advantage of individual quantifications of several species of microcystins.

키랄 크로마토그래피와 NMR 분광법에 의한 N-acetyl-1-naphthylethylamine의 광학순도 측정 (1Determination of optical purity of N-acetyl-1-naphthylethylamine by chiral chromatography and NMR spectroscopy)

  • 정영한;류재정
    • 분석과학
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    • 제23권1호
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    • pp.97-101
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    • 2010
  • (R)-N-3,5-dinitrobenzoyl (DNB) phenylglycinol로 부터 만들어진 키랄 선택제가 라세미 Nacylnaphthylalkylamines의 분리에 HPLC 키랄 정지상으로 이용된 바 있다. 본 연구에서는 (R)-phenylglycinol 유도체 키랄 선택제를 이용하여 키랄 크로마토그래피와 NMR 분광법에 의한 광학순도를 측정하였다. NMR과 HPLC 실험결과를 참값과 비교하여 각 광학순도 측정값의 정확도와 정밀도를 계산하였다. NMR 방법의 오차는 +2.2%, 평균 RSD는 4.54% 이었고, HPLC 방법의 오차는 -3.5%, 평균 상대표준편차는 3.23% 이었다.

A single-step isolation of useful antioxidant compounds from Ishige okamurae by using centrifugal partition chromatography

  • Kim, Hyung-Ho;Kim, Hyun-Soo;Ko, Ju-Young;Kim, Chul-Young;Lee, Ji-Hyeok;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • 제19권4호
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    • pp.22.1-22.7
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    • 2016
  • One of the main compounds in Ishige okamurae, diphlorethohydroxycarmalol (DPHC), is known to exhibit antiviral and anti-inflammatory effects. However, it has not been investigated extensively. In this study, preparative centrifugal partition chromatography (CPC) coupled with 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) online HPLC was employed for effectively separating considerable amounts of antioxidant compounds from marine algae. Two main antioxidant compounds, DPHC and octaphlorethol A (OPA), respectively, were confirmed and isolated from the ethyl acetate (EtOAc) fraction of I. okamurae by $ABTS^+$ online HPLC and preparative CPC systems. The presence of DPHC and OPA was confirmed in the EtOAc fraction of I. okamurae by both liquid chromatography with diode array detection and electrospray ionization mass spectrometry (LC-DAD-ESI/MS) and $ABTS^+$ online HPLC systems: DPHC (39 mg) and OPA (23 mg) were successfully isolated from I. okamurae (500 mg) with optimum solvent composition (0.5:10:4:6; n-hexane/EtOAc/MeOH/water, v/v) with corresponding partition coefficients (K) of 1.62 and 2.71, respectively, by preparative CPC. Hence, CPC coupled with $ABTS^+$ online HPLC is convenient for the efficient and simple isolation of these antioxidant compounds from I. okamurae.

전통된장으로부터 Angiotensin Converting Enzyme 저해물질의 분리 (Isolation of Angiotensin Converting Enzyme Inhibitor from Doenjang)

  • 김승호;이윤진;권대영
    • 한국식품과학회지
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    • 제31권3호
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    • pp.848-854
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    • 1999
  • 전통된장으로부터 안지오텐신전환효소(angiotensin converting enzyme; ACE)를 저해하는 물질을 추출하여 그 구조를 밝혀냈다. ACE 저해물질을 열수추출한 다음 gel permeation chromatography (GPC)를 통하여 ACE 저해작용이 큰 두 개의 큰 획분을 받았다. 앞획분은 90%와 70%의 ACE 저해효과를 나타내었으나 단일물질로 분리되지 않아 계속하여 역상 HPLC를 통하여 순수 분리를 하였다. 그러나 앞획분은 순수분리되지 않아 결국 2차원 전기영동/TLC를 통하여 분리한 결과 분자량이 759.63인 아미노기를 갖고 있는 비펩타이드 물질임이 밝혀졌다. 뒷획분은 다른 조건의 HPLC(reverse column과 $NH_2$, column)를 이용하여 순수분리에 성공하였다. 이중 ACE 저해효과가 큰 물질은 분자량 271.33인 dipeptide인 arginine-proline임을 밝혀냈다. 이물질의 ACE $IC_{50}$$92\;{\mu}M$이었다. 본 연구 결과는 대부분 ACE 저해물질이 3개 내지 7개 등의 긴 펩타이드임을 감안할 때, 짧은 dipeptide로 ACE 저해펩타이드가 한국의 전통된장에서 생산할 수 있음을 보여주고 있다.

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일반계 벼의 brassinosteroid활성물질 검색 (Investigation of brassinosteroid substances in the shoots of japonica type Korean rice)

  • 현규환;박근형
    • Applied Biological Chemistry
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    • 제33권4호
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    • pp.307-314
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    • 1990
  • Brassinosteroid는 특이한 생리작용을 갖는 식물의 신 생장조절물질로 주목을 받고 있으나, 아직 우리나라의 가장 중요한 작물인 벼의 brassinosteroid에 대해 연구된 바 없어, 일반계 품종의 벼를 대상으로 brassinosteroid활성물질의 검색을 시도하였다. 동진벼와 영산벼를 최고분얼기까지 재배하여 지상부를 수확하고, 유기용매로 추출하여 얻어진 추출물에서 rice lamina inclination test에 의한 생물검정법으로 활성이 인정되어, 활성본체의 구명과 정제의 목적으로 각각의 활성획분을 silica gel 흡착 chromatography, Sephadex LH-20 chromatography, charcoal 흡착 chromatography, preparative TLC, Bondesil chromatography 그리고 normal, reverse phase의 HPLC 등적 방법으로 분획하여 검정한 결과 2품종 모두 brassinosteroid활성물질을 생산하고 있으며, 또 2품종간의 endogenous brassinosteroid의 유사성이 인정되었다.

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밤생율(生栗)에 함유된 Peroxidase의 정제 및 특성에 관한 연구 (Purification and Properties of the Peroxidase in Castanea Semen)

  • 오석홍;김용휘;이서나
    • 한국식품과학회지
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    • 제19권6호
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    • pp.506-514
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    • 1987
  • 밤(생율(生栗))으로 부터 peroxidase를 $(NH_4)_2SO_4$에 의한 염석 및 DEAE-cellulose column chromatography, sephadex G-100 column chromatography, HPLC방법으로 정제하였으며 정제도는 조효소에 비하여 65.3배였고, HPLC로 측정한 밤 peroxidase 분자량은 35,000으로 추정되었다. sephadexG-100 column chromatography 후 얻은 밤 peroxidase의 작용최적 pH는 5.0이었고, 작용최적 온도는 $50^{\circ}C$이었으며 $80^{\circ}C$에서 1.73분 열처리할 때 90%의 효소가 불활성화되었다. 본 효소는 OPDA 및 PPDA와 같은 방향족 amine류에 높은 활성을 나타내었다. OPDA와 $H_2O_2$에 대한 Km치는 각각 2.6mM과 10mM이었고, 저해작용은 L-ascorbic acid 와 sodium L-ascobate가 가장 컸으며, $Ca^{++}$$Ba^{++}$은 1mM과 5mM에서 현저히 효소활성을 증가시켰다.

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Development of Pretreatment Method for Analysis of Vitamin B12 in Cereal Infant Formula using Immunoaffinity Chromatography and High-Performance Liquid Chromatography

  • Park, Jung Min;Koh, Jong Ho;Kim, Jin Man
    • 한국축산식품학회지
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    • 제41권2호
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    • pp.335-342
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    • 2021
  • Vitamin B12 deficiency may lead to serious health issues in both infants and adults. A simple analytical method involving sample pretreatment with enzyme, followed by cyanide addition under acidic conditions; separation on an immunoaffinity column; and high-performance liquid chromatography (HPLC) was developed for the rapid detection and quantitation of vitamin B12 in powdered milk. Detection limit and powdered milk recovery were determined by quantitative analysis. The limits of detection and quantitation were 2.71 and 8.21 ㎍/L, respectively. Relative standard deviations of the intra-day and inter-day precisions varied in the ranges of 0.98%-5.31% and 2.16%-3.90%, respectively. Recovery of the analysis varied in the range of 83.41%-106.57%, suggesting that the values were acceptable. Additionally, vitamin B12 content and recovery in SRM 1849a were 54.10 ㎍/kg and 112.24%, respectively. Our results suggested that the analytical method, including the sample pretreatment step, was valid. This analytical method can be implemented in many laboratory-scale experiments that seek to save time and labor. Therefore, this study shows that immunoaffinity-HPLC/ultraviolet is an acceptable technique for constructing a reliable database on vitamin B12 in powdered milk containing starch as well as protein and/or fat in high amounts.