• Title/Summary/Keyword: salting-out extraction

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

  • Woong Hur;Chang-Lyuk Yoon
    • Journal of Oral Medicine and Pain
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    • v.20 no.2
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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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    • v.6 no.5
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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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    • v.44 no.1
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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
    • Analytical Science and Technology
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    • v.8 no.4
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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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Determination of $Mg^{2+}$ using 5-membered Heterocyclic Compound (5원자 헤테로고리 화합물을 이용한 $Mg^{2+}$의 정량)

  • Seo, Moo Lyong;Lee, Shim Sung;Kim, Jae Sang;Park, Tae Myung
    • Analytical Science and Technology
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    • v.6 no.3
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    • pp.307-311
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    • 1993
  • Ionophore, which contains 5-membered heterocyclic compound, was prepared. $Mg^{2+}$ was determined by salting-out technique using ionophore as a chelating reagent. After $Mg^{2+}$ was extracted into the acetonitrile layer as a Mg-$(Ionophore)_2$ complex from acetate buffered aqueous solution by salting-out extraction technique, absorbance of complex was recorded by atomic absorption spectrophotometry. Optimum pH was between 2.5 and 5.0 for extraction and 1:2([$Mg^{2+}$]/[ionophore]) complex were formed. The range of detection was 0.24ppm~2.4ppm and $Ca^{2+}$ and EDTA were interfered in the determination of $Mg^{2+}$.

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

  • Lee, Sanghoi;Song, Jaesok;Yoon, youngshik;Kim, Chinyon;Won, Jonguk;Roh, Jaehoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.10 no.1
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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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Determination of Mn, Co, Ni and Cu in Iron Oxide Ore by Atomic Absorption spectroscopy. Utilization of APDC-MIBK Extraction System (원자흡수 분광법에 의한 철광석중의 Mn, Co, Ni 및 Cu 의 정량. APDC-MIBK 추출계의 이용)

  • Misun Park;Youn-Doo Kim;Kwanghee Koh Park
    • Journal of the Korean Chemical Society
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    • v.33 no.3
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    • pp.315-320
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    • 1989
  • A method was presented for the analysis of trace metals in iron oxide ore. The method utilized ammonium pyrrolidinedithiocarbamate (APDC)-methyl isobutyl ketone (MIBK) extraction procedure and analysis by atomic absorption spectroscopy (AA). Citrate at pH $8{\sim}10$ for the determination of Co, Ni and Cu or tiron at pH $6{\sim}7$for the determination of Mn and Cu was added as a masking agent to prevent extraction of Fe(III) into the organic phase. Reduction of solubility of MIBK in water was achieved by addition of NaCl as a salting-out agent. Back extraction of the MIBK extracts with aqueous $HNO_3$ was also studied to increase the stability of metal extracts.

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

  • Hong, Yeon Ki
    • Korean Chemical Engineering Research
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    • v.51 no.5
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    • pp.575-579
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    • 2013
  • 1,3-Propandiol is a promising chemical which can be produced from fermentation of glycerol because the application of 1,3-propanediol is mainly in the production of bio-polytrimethylene terephthalate (bio-PTT). However, the cost of downstream processes in the biological production of 1,3-propanediol can make a high portion in the total production cost due to the large amount of water and the by-produced carboxylic acids such as succinic, lactic and acetic acids in 1,3-propanediol fermentation broth. In this study, aqueous two-phases systems composed of hydrophilic alcohols and phosphate salts were applied to the recovery of 1,3-propanediol from its artificial aqueous solution. Formation of aqueous biphases in hydrophilic alcohols and phosphate salts was due to the salting-out effect of salts in bottom phase, thereby 1,3-propanediol in bottom phase was moved into top phase. Extraction efficiency for 1,3-propanediol was proportional to the polarity of hydrophilic alcohols and the basicity of salts and the maximum value of extraction efficiency was more than 98%. In the aqueous two-phases systems after extraction, there was no carboxylic acid in top phase. Therefore, it was concluded that the aqueous two-phases systems composed of hydrophilic alcohols and phosphate salts were effective for the selective recovery of 1,3-propanediol without any coextraction of carboxylic acids.

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

  • Kang, Tae-Woo;Pyo, Hee-Soo;Hong, Jong-Ki
    • Analytical Science and Technology
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    • v.21 no.4
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    • pp.259-271
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    • 2008
  • The analytical method of extracting compounds from human blood to examine accumulated organochlorine pesticides (OCPs) has been widely used the traditional liquid-liquid extraction (LLE) method and solid-phase extraction (SPE) method, yet these methods have certain limitations in purification and usafe of a large amount of sample. In order to overcome the se problems reside in these, solid-phase microextraction (SPME), known as a highly efficient extration method with less samples and relatively simple, was employed to collect 18 different kinds of OCPs in blood as extraction method in this study. To optimize extraction method, we examine various experimental SPME-parameters such as adsorption (fiber type, adsorption time, adsorption temperature, salting out effect), and desorption (desorption time, desorption temperature etc.). From the experimental results, the optimal conditions are as follows: fiber was polyacrylate with $85{\mu}m$, adsorption time was for 5 min, adsorption optimum temperature was at $280^{\circ}C$, and salting out effect was NaCl with 0.1 g. MDL, precision and accuracy was in the ranges of 0.05~0.20 ng/mL, 5.59~13.39%, respedively, and accuracy was -0.5% ~24.5% for all OCPs.

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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    • v.34 no.11
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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.