• 제목/요약/키워드: salting-out extraction

검색결과 20건 처리시간 0.034초

치아를 이용한 성별검사 및 D1S80 유전좌위의 검색시 4가지 DNA추출방법에 따른 비교 (Comparison of 4 Methods of DNA Extraction for Sex Determination and D1S80 Locus Detection in Teeth)

  • Woong Hur;Chang-Lyuk Yoon
    • Journal of Oral Medicine and Pain
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    • 제20권2호
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    • pp.497-513
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    • 1995
  • Human genomic Deoxyribonucleic acid(DNA) was extracted from teeth by boiling, salting-out, phenol, boiling-phenol methods. The author compared DNA concentration and its purity, the accuracy of sex determination and the results of the D1S80 locus detection among above 4 methods. The following results were obtained : 1. DNA concentration was the highest in pulp with salting-out method and DNA purity was higher in pulp with salting-out and phenol methods than other 2 methods. 2. Sex determination was possible using of the pulp and the dentin of the teeth with four methods but, it was impossible in the enamel and some pulp with boiling method. 3. Amplification of D1S80 locus occurred from pulp and dentin with salting-out, phenol, and boiling-phenol methods. 4. There are no differences among the amplification of X-Y homologus amelogenin gene by application of 4 methods and salting-out, phenol methods efficiently makes available to amplification of D1S80 locus. From the investigation DNA extraction, sex determination, amplification of D1S80 locus was successfully accomplished with salting-out, phenol, boiling-phenol methods Therefore above 3 methods are available and applicable as forensic odontology for individual identification.

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Salting-out Effects on the Partition of Proteins in Aqueous Two-phase Systems

  • KIM, CHAN-WHA;CHO KYUN RHA
    • Journal of Microbiology and Biotechnology
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    • 제6권5호
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    • pp.352-357
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    • 1996
  • The partition of proteins in the salt-rich phase of polyethylene glycol (pEG)/salt aqueous two-phase systems is limited by the salting-out effects of salt. The logarithm of the concentration of proteins partitioned in the salt-rich phase decreases linearly with increases in the concentration of salt in the salt-rich phase (salting-out). Therefore, the partition of a given protein in the salt-rich phase of aqueous two-phase systems can be estimated from the salting-out constant. The slope of the solubility line (salting-out con-stant) for a given protein is determined by the type of salt in the two-phase systems.

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Salting-out extraction of ginsenosides from the enzymatic hydrolysates of Panax quinquefolium based on ethanol/sodium carbonate system

  • Wei, Yingqin;Hou, Baojuan;Fang, Haiyan;Sun, Xinjie;Ma, Feng
    • Journal of Ginseng Research
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    • 제44권1호
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    • pp.44-49
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    • 2020
  • Background: Salting-out extraction (SOE) had been developed as a special branch of aqueous two-phase system recently. So far as we know, few reports involved in extracting ginsenosides with SOE because of the lower recovery caused by the unique solubility and surface activity of ginsenosides. A new SOE method for rapid pretreatment of ginsenosides from the enzymatic hydrolysates of Panax quinquefolium was established in this article. Methods: The SOE system comprising ethanol and sodium carbonate was selected to extract ginsenosides from the enzymatic hydrolysates of Panax quinquefolium, and HPLC was applied to analyze the ginsenosides. Results: The optimized extraction conditions were as follows: the aqueous two-phase extraction system comprising ethanol, sodium carbonate, ethanol concentration of 41.51%, and the mass percent of sodium carbonate of 7.9% in the extraction system under the experimental condition. Extraction time had minor influence on extraction efficiency of ginsenosides. The results also showed that the extraction efficiencies of three ginsenosides were all more than 90.0% only in a single step. Conclusion: The proposed method had been successfully applied to determine ginsenosides in enzymatic hydrolysate and demonstrated as a powerful technique for separating and purifying ginsenosides in complex samples.

Extraction of Water-Soluble Porphyrin and Metalloporphyrins into Acetonitrile by Salting-out

  • Tabata, Masaaki;Kumamoto, Midori
    • 분석과학
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    • 제8권4호
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    • pp.511-517
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    • 1995
  • A cationic water soluble porphyrin (5,10,15,20-tetrakis (l-methyl-pyridinium-4-yl)porphyrin, $H_2tmpyp^{4+}$) and its metalloporphyrins (MP) were easily extracted into acetonitrile separated by addition of sodium chloride ($4mol\;dm^{-3}$) in the presence of sodium perchlorate, where M denotes $Zn^{2+}$, $Cu^{2+}$, $Co^{3+}$, $Fe^{3+}$, and $Mn^{3+}$ and $P^{2-}$ is porphyrinate ion. The extracted ion-pair complexes were completely dissociated to $[MP(ClO_4)_3]^+$, and $[MP(ClO_4)_2]^{2+}$. The extraction and the dissociation constants were determined by taking into account of the partition constant of sodium perchlorate ($K_D=1.82{\pm}0.01$). The chemical properties of the separated acetonitrile phase as $E_{T(30)}$ and $D_{II,I}$ were determined and compared with other water miscible solvents (acetone, actonitrile, 1,4-dioxane, tetrahydrofuran, 1-propanol and 2-propanol). Furthermore, a sensitive and selective method was proposed for the determination of a subnanogram amount of copper(II) in natural water samples by using the present salting-out method and the porphyrins.

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5원자 헤테로고리 화합물을 이용한 $Mg^{2+}$의 정량 (Determination of $Mg^{2+}$ using 5-membered Heterocyclic Compound)

  • 서무룡;이심성;김재상;박태명
    • 분석과학
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    • 제6권3호
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    • pp.307-311
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    • 1993
  • 5원자 헤테로고리를 포함하는 열린고리형의 ionophore를 합성하였다. 이 때 합성한 ionophore를 착화제로 이용하여 수용액 중의 $Mg^{2+}$을 염석추출법으로 추출하여 원자 흡광 광도법으로 정량하였다. Acetate 완충용액으로 pH를 4.2로 조절한 $Mg^{2+}$를 포함하는 수용액에 ionophore-AN 용액을 넣어 $Mg^{2+}$-ionophore 착물을 형성시킨 다음, 염석제를 첨가하여 유기층으로 추출하고, 유기층의 착물의 흡광도를 원자 흡광 광도계로서 측정하여 $Mg^{2+}$를 정량하였다. 최적 추출 조건을 조사해 본 결과, 최적 pH는 2.5~5.0이었으며, 염석제인 황산암모늄[$(NH_4)_2SO_4$]의 양은 5g 이상이면 정량적으로 추출되었다. 또한 $Mg^{2+}$과 ionophore의 착물 조성비는 1:2이었다. $Mg^{2+}$의 정량범위는 0.24ppm~2.4ppm이었으며 $Ca^{2+}$이온 및 EDTA의 방해효과가 크게 나타났다.

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부타놀 추출법을 이용한 2-thiothiazolidine-4-carboxilic acid와 thiocarbamide의 동시정량에 관한 연구 (Simultaneous analysis for 2-thiothiazolidine-4-carboxilic acid and thiocarbamide using butanol extraction method)

  • 이상회;송재석;윤영식;김치년;원종욱;노재훈
    • 한국산업보건학회지
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    • 제10권1호
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    • pp.208-222
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    • 2000
  • This study was conducted to supplement limit of previous study, The objectives of this study were to select optimal conditions of high performance liquid chromatography(HPLC) operation for detecting urinary 2-thiothiazolicline-4-carboxylic acid(TTCA) and thiocarbamide simultaneously, and to evaluate recovery rates for various liquid-liquid extration method of these metabolites, The results are as follows : 1. The urinary TTCA and thiocarbamide were separate sharply when flow rate is $0.7m{\ell}/min$, using a series $C_8$ and $C_{18}$ column, 50 mM $KH_2PO_4$ : acetonitrile (93.5 : 6.5) and pH 3.5 as a mobile phase. The retention time was TTCA, $12.07{\pm}0.11$(mean${\pm}$SD, n=06), thiocarbamide, $7.85{\pm}0.01$ (mean${\pm}$SD, n=6), respectively. The calibration curve for TTCA and thiocarbamide was linear within the range 0.05 to $30{\mu}g/m{\ell}$. 2. By the liquid-liquid extration, butanol extration with $(NH_4)_2$ as a salting-out reagent was used as a simultaneous extration method for these metabolites in acid state, and recovery rates of this method are urinary TTCA, $49.6{\pm}17.7$ (mean${\pm}$SD, n=16), thiocarbamide, $43,9{\pm}5.50$ (mean${\pm}$SD, n=16), respectively 3. The precision(pooled coefficients of variation for 4 concentration) of the urinary thiocarbamide analysis was 0.03754 by butanol liquid-liquid extraction with $(NH_4)_2$ as a salting-out reagent, and TTCA was 0.04082 by ethyl acetate liquid-liquid extration with $(NH_4)_2$ as a salting out reagent The above results show that the butanol liquid-liquid extraction with $(NH_4)_2$ as a salting-out reagent in acid state, and using a series $C_8$ and $C_{18}$ column, 50 mM $KH_2PO_4$ : acetonitrile (93.5 : 6.5) and pH 3.5 as a mobile phase are suitable for the analysis of urinary TTCA and thiocarbamide simultaneously. The detection limit of TTCA and thiocarbamide was about $0.17{\mu}g/m{\ell}$, $0.07{\mu}g/m{\ell}$.

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원자흡수 분광법에 의한 철광석중의 Mn, Co, Ni 및 Cu 의 정량. APDC-MIBK 추출계의 이용 (Determination of Mn, Co, Ni and Cu in Iron Oxide Ore by Atomic Absorption spectroscopy. Utilization of APDC-MIBK Extraction System)

  • 박미선;김연두;고광희
    • 대한화학회지
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    • 제33권3호
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    • pp.315-320
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    • 1989
  • 철광석 중에 존재하는 미량 금속원소들을 분석하는 방법을 제시하였다. 분석방법은 ammonium pyrrolidinedithiocarbamate(APDC)-methyl isobutyl ketone(MIBK) 추출계와 원자흡수 분광법(AA)를 이용하였다. 철이 유기층으로 추출되는 것을 막기 위하여 가리움제로서 코발트, 니켈 및 구리의 분석시에는 pH $8{\sim}10$에서 citrate를, 그리고 망간 및 구리의 분석시에는 pH 6{\sim}7$에서 tiron을 가하였다. MIBK의 물에 대한 용해도를 줄이기 위해 NaCl을 염석제로 가하였다. 금속 추출용액의 안정도를 증가시키기 위한 방안으로 MIBK 금속 추출물을 질산 수용액으로 역추출하는 것을 검토하였다.

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수상이성분계에 의한 1,3-프로판디올 추출특성에 대한 알콜과 카르복실산의 영향 (Effect of Alcohols and Carboxylic Acids on Extraction Characteristics for 1,3-Propanediol by Aqueous Two Phases Systems)

  • 홍연기
    • Korean Chemical Engineering Research
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    • 제51권5호
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    • pp.575-579
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    • 2013
  • 최근 바이오 플라스틱인 Polytrimethylene terephthalate (PTT)의 원료물질인 1,3-프로판디올의 생물학적 생산이 주목받고 있다. 발효를 통한 1,3-프로판디올의 생산에 있어 부산물로 생성되는 젖산, 숙신산을 포함한 카르복실산들은 1,3-프로판디올의 경제적인 생산을 어렵게 한다. 본 연구에서는 카르복실산 부산물이 포함된 1,3-프로판디올 수용액으로부터 1,3-프로판디올의 효율적 분리정제를 위해 수용성 알콜과 염으로 구성된 수상이성분계를 이용한 추출을 적용하였다. 알콜과 염으로 구성된 수상이성분계 형성은 하부상에 존재하는 염의 염출효과에 의한 것이며 이로 인해 1,3-프로판디올이 상부상으로 이동된다. 추출효율을 알콜의 사슬길이가 짧을수록, 염의 세기가 높을수록 높아졌으며 최대 98%의 회수율을 얻을 수 있었다. 또한 발효 시에 부산물로 형성되는 카르복실산은 1,3-프로판디올과 동반추출 되지 않았으며 1,3-프로판디올의 추출효율에도 영향이 없는 것으로 나타났다. 그러므로 본 연구에서 적용된 친수성 알콜과 염으로 구성된 수상이성분계는 카르복실산 부산물로부터 1,3-프로판디올을 선택적으로 분리하는데 효과적임을 알 수 있다.

HS SPME-GC/MS를 이용한 혈액 중 유기염소계 농약의 분석법 개발 (Development of an analytical method of organochlorine pesticides in human bloods using head space-solid phase microextraction coupled with gas chromatography/mass spectrometry)

  • 강태우;표희수;홍종기
    • 분석과학
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    • 제21권4호
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    • pp.259-271
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    • 2008
  • 혈액 내에 축적되어 있는 유기염소계 농약을 분석하기 위한 방법으로 기존에는 액체-액체 추출법과 고체상 추출법을 많이 사용하였으나 정제 및 많은 양의 시료 처리에 한계가 있다. 이를 극복하기 위한 방법으로 시료양이 적으며 간편한 고체상미량추출법을 사용하여 혈액 중 18종의 유기염소계 농약의 최적 추출법을 연구하였다. 흡착조건(fiber type, 흡착시간, 흡착온도, 염석효과)과 탈착조건(탈착시간, 탈착온도) 등 최적의 추출을 위해 여러 가지의 실험조건 등을 검토한 결과, fiber는 polyacrylate $85{\mu}m$, 흡착시간은 50분, 흡착온도는 $80^{\circ}C$, 염석효과는 NaCl 0.1 g, 탈착시간은 5분, 탈착온도는 $280^{\circ}C$에서 최적임을 확인하였다. 정확도, 정밀도 및 검출한계에 대한 타당성을 최적 실험조건에서 조사한 결과, 검출한계는 0.05~0.20 ng/mL, 정밀도는 5.59~13.39%, 정확도는 -0.5%~24.5%의 범위인 것으로 확인되었다.

Determination of Non-Steroidal Anti-Inflammatory Drugs in Human Urine Sample using HPLC/UV and Three Phase Hollow Fiber-Liquid Phase Microextraction (HF-LPME)

  • Cha, Yong Byoung;Myung, Seung-Woon
    • Bulletin of the Korean Chemical Society
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    • 제34권11호
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    • pp.3444-3450
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    • 2013
  • Three phase hollow fiber-liquid phase microextraction (HF-LPME), which is faster, simpler and uses a more environmentally friendly sample-preparation technique, was developed for the analysis of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) in human urine. For the effective simultaneous extraction/concentration of NSAIDs by three phase HF-LPME, parameters (such as extraction organic solvent, pH of donor/acceptor phase, stirring speed, salting-out effect, sample temperature, and extraction time) which influence the extraction efficiency were optimized. NSAIDs were extracted and concentrated from 4 mL of aqueous solution at pH 3 (donor phase) into dihexyl ether immobilized in the wall pores of a porous hollow fiber, and then extracted into the acceptor phase at pH 13 located in the lumen of the hollow fiber. After the extraction, 5 ${\mu}L$ of the acceptor phase was directly injected into the HPLC/UV system. Simultaneous chromatographic separation of seven NSAIDs was achieved on an Eclipse XDB-C18 (4.6 mm i.d. ${\times}$ 150 mm length, 5 ${\mu}m$ particle size) column using isocratic elution with 0.1% formic acid and methanol (30:70) at a HPLC-UV/Vis system. Under optimized conditions (extraction solvent, dihexyl ether; $pH_{donor}$, 3; $pH_{acceptor}$, 13; stirring speed, 1500 rpm; NaCl salt, 10%; sample temperature, $60^{\circ}C$; and extraction time, 45 min), enrichment factors (EF) were between 59 and 260. The limit of detection (LOD) and limit of quantitation (LOQ) in the spiked urine matrix were in the concentration range of 5-15 ng/mL and 15-45 ng/mL, respectively. The relative recovery and precision obtained were between 58 and 136% and below 15.7% RSD, respectively. The calibration curve was linear within the range of 0.015-0.96 ng/mL with the square of the correlation coefficient being more than 0.997. The established method can be used to analyse of NSAIDs of low concentration (ng/mL) in urine.