• Title/Summary/Keyword: Salting-out

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A New Microassay for the Determination of Serum TBA (Thiobarbituric Acid) Value Enhanced by $FeCl_{3}$ under Anaerobic Condition (무산소상태에서 $FeCl_{3}$로 촉진된 Thiobarbituric Acid 반응을 이용한 혈청중 TBA 반응물의 새로운 정량법)

  • Lee, Joung-Won;Mo, Su-Mi;Lee, Tae-Young
    • Journal of Nutrition and Health
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    • v.17 no.1
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    • pp.31-40
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    • 1984
  • A new microassay was proposed for the determination of serum thiobarbituric acid(TBA) value greatly enhanced by ferric ion under anaerobic condition. One ${\mu}mole$ of $FeCl_{3}$ per $10{\mu}l$ of serum was added to the TBA reaction mixture containing serum protein precipitate. The reaction mixture was heated on boiling water-bath for 50min under $N_{2}$ flushing. The sensitivity of this assay was greatly enhanced by 40 times comparing with that of Yagi's method (1976). In favour of the enhancement, this test could be measured by colorimetry or spectrophotometry with the sample size of $10-20{\mu}l$ serum. The sensitivity and reproducibility were also improved by means of partial dehydration of the butanol extract with $(NH_{4})_{2}SO_{4}$ salting-out. Serum TBA values of healthy human at different age groups were determined by this proposed method.

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Study on purification and extraction of nitrate salts from waste scrubbing liquid of de-SOx/de-NOx (탈질/탈황 폐 세정액으로부터 질산염 추출 및 정제 연구)

  • Kim, Woo-Ram;Jo, Young-Min;Lee, Heon-Seok;Oh, Soo-Kwan
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.1
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    • pp.48-55
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    • 2015
  • IMO to issue some restricted maritime legislation for reducing the adverse environmental impacts arising from such ship exhaust emissions. According to the IMO policy, every ship entering the Baltic SECAs has to equip the gas cleaning scrubber. The discharged waste solution by gas cleaning scrubber contains many types of salts, which to recover some valuable materials before disposal. This study try to achieve valuable salts including AN and AS throughout a few process such as selective organic solvents salting out, low temperature extraction and thermal evaporation. Amongst them, Thermal evaporation with repetition extraction using inorganic solvent was the most optimum to purify the extracted AN. This valuable salt was evaluated by Elemental analysis and Differential scanning calorimetry.

Study on the Fouling Reduction of the RO Membrane by the Coating with an Anionic Polymer (역삼투막 표면에 음이온 고분자 코팅을 통한 파울링 현상 감소연구)

  • Cho, Eun Hye;Cheong, Seong Ihl;Rhim, Ji Won
    • Membrane Journal
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    • v.22 no.6
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    • pp.481-488
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    • 2012
  • The anionic exchange polymer, poly(vinyl amine)(PVAm), was coated onto polyamide (PA) reverse osmosis (RO) membranes by using 'salting-out' method. The effects of the fouling phenomena for these PVAm coated membranes were investigated using the model foulants, bovine serum albumin (BSA), humic acid (HA), and sodium alginate (SA). The surface coating and the fouling phenomena were observed by the scanning electron microscopy. And the flux was measured for each 100 ppm of above foulant aqueous solution at the operating pressure, 2, 4, 8 bar. The PVAm-coated PA membranes showed somewhat fouling improvements and the fouling reduction was shown in the order of HA > SA > BSA, particularly HA case was distinct.

Isolation and production of soymilk-clotting enzymes from Bacillus sp. K-324-7 (대두유 응고효소 생산에 관한 연구)

  • Lee, Gi-Soung;Han, Myun-Soo;Shim, Sang-Kook;Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.33 no.2
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    • pp.154-160
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    • 1990
  • A bacterial strain which was capable of producing extracellular soymilk-clotting enzyme was isolated from soil samples during the course of screening test. The characteristics of the isolated strain K-324-7, indicated that the strain belonged to species of Bacillus cereus. The crude purification of this enzyme was precipitated by salting out with ammonium sulfate of 0.8 saturation. The optimal pH for the enzyme activity was at $6.1{\sim}7.0$ and below $50^{\circ}C$. The optimal culture medium for the production of soymilk-clotting enzyme were consisted of 0.2% glucose, 0.2% peptone, and 0.5% $KH_2PO_4$ with initial pH value of 6.5. The activity of enzyme was maximum when the microbe was cultured for 3 days at $35^{\circ}C$.

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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.

Purification of Intracellular $\beta$-Galactosidase from Lactobacillus sporogenes in an Aqueous Poly(ethylene glycol)- Potassium Phosphate Two-Phase System (Poly(ethylene glycol)/인산염 용액 2상계를 이용한 Lactobacillus sporogenes가 생산하는 균체내 $\beta$-Galactosidase의 추출 분리에 관한 연구)

  • 이삼빈;김영만;이철호
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.84-88
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    • 1987
  • Poly(ethylene glycol)-PPB two phase system was used tot the purification of $\beta$-galactosidase from Lactobacillus sporogenes. The smaller the molecular weight of concentration of PEG phase in-creased, proteins as well as $\beta$-galactosidase was partitioned into the top phase. All cell debris were confined to the potassium phosphate phase (bottom phase), approached to the binodial line. The purification ratio increased by changing the polymer-salt composition of the tie line towards higher salt concentrations. It was also possible to obtain higher purification of the enzyme after two-step extraction using PEG 1000 and PEG 300. The top phase contained 74% of the total $\beta$-galactosidase with a purification factor of 2.1.

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Receptor activator of nuclear factor kappa-B gene polymorphisms in Iranian periodontitis and peri-implantitis patients

  • Kadkhodazadeh, Mahdi;Baghani, Zahra;Ebadian, Ahmad Reza;Kaghazchi, Zahra;Amid, Reza
    • Journal of Periodontal and Implant Science
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    • v.44 no.3
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    • pp.141-146
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    • 2014
  • Purpose: Peri-implantitis and periodontitis are inflammatory and infectious diseases of implant and tooth-supporting tissues. Recently, the role of gene polymorphisms of immune response components in the relevant pathogenesis has been investigated. The present study was the first to evaluate the relationship between two known single nucleotide polymorphisms (SNPs) of the receptor activator of nuclear factor kappa-${\beta}$ (RANK) gene (rs3018362 and rs35211496) in chronic periodontitis and peri-implantitis patients in an Iranian population. Methods: Eighty-one periodontally healthy patients, 38 patients with peri-implantitis, and 74 patients with chronic periodontitis were enrolled in this study. DNA was extracted from blood arm vein samples by using Miller's salting out technique according to the manufacturer's instructions given in the extraction kit. The concentration of DNA samples was measured using a spectrophotometer. The genetic polymorphisms of the RANK gene were evaluated using a competitive allele specific polymerase chain reaction (KBioscience allele specific PCR) technique. Differences in the frequencies of genotypes and alleles in the diseased and healthy groups were analyzed using chi-squared statistical tests (P<0.05). Results: Analysis of rs35211496 revealed statistically significant differences in the expression of the TT, TC, and CC genotypes among the three groups (P=0.00). No statistically significant difference was detected in this respect between the control group and the chronic periodontitis group. The expression of the GG, GA, and AA genotypes and allele frequencies (rs3018362) showed no statistically significant difference among the three groups (P=0.21). Conclusions: The results of this study indicate that the CC genotype of the rs35211496 RANK gene polymorphism was significantly associated with peri-implantitis and may be considered a genetic determinant for peri-implantitis, but this needs to be confirmed by further studies in other populations.

Production ani Some Properties of Milk Clotting Enzyme from Mucor sp. (Mucor sp. 에 의한 응유효소생산(凝乳酵素生産)과 그의 성질(性質)에 관하여)

  • Yeum, Dong Kil;Kim, Chan Jo;Lee, Jong Soo
    • Korean Journal of Agricultural Science
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    • v.14 no.1
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    • pp.144-155
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    • 1987
  • A potent fungus producing milk clotting enzyme with fairly weak proteolytic activity was isolated from various soil and sewage, which the selected strain, SA-101, was identified as Mucor sp. with microbiological characteristics. Its milk clotting enzyme production was maximized when grown on 10g of wheat bran media added to 8ml of tap water containing 0.1M HCl for 60hrs at $30^{\circ}C$. This enzyme production was stimulated by addition of 6% lactose, 0.05% NaCl and reached a maximal level of 9810 unit/g wheat bran. The crude enzyme product could be produced effectively by salting out with ammonium sulfate fractionation and lyophilization. The ratio of milk clotting activity to proteolytic activity of crude enzyme product was lower than Hansen rennet, but resembled to Meito rennet. The optimal temperature of milk clotting activity of crude enzyme product was abound $60^{\circ}C$ on a substrate of 10% reconstituted skim milk containing 1/100M $CaCl_2$.

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Studies on the Lipolytic Enzyme of Molds -Part 3. Purification of Rhizopus japonicus lipase- (사상균(絲狀菌)의 지방분해효소(脂肪分解酵素)에 관(關)한 연구(硏究) -제3보(第3報) 분리사상균(分離絲狀菌) Rhizopus japonicus lipase의 정제(精製)에 관(關)하여)

  • Chung, Man-Jae
    • Korean Journal of Food Science and Technology
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    • v.8 no.3
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    • pp.141-146
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    • 1976
  • 1) The purified enzyme was obtained with the specific activity 126.5 u/mg protein (about 45 times of the original activity) and the yield of 4.2%, by means of salting out with ammonium sulfate (0.5 saturation) of the crude enzyme solution, desalting by Sephadex G 25, CM cellulose columm chromatography, concentration by Sephadex G 25, and gel filtration by Sephadex G 75. 2) In the acrylamide gel disc electrophoresis of the purified enzyme, the main band and two obscure ones on the both side of the main band appeared, which indicated that the enzyme was considerably purified compared with its crude enzyme solution, even if it is not referred to as a pure protein.

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Further induction of amylase producing mutants from a highly proteolytic mutant strain of asppergillus flavus (돌연변이에 의한 Aspergillus flavus의 아밀라아제 생성능의 개량)

  • 이영록;고상균;김봉수
    • Korean Journal of Microbiology
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    • v.18 no.4
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    • pp.161-171
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    • 1980
  • A mutant strain having increased productivity of both enzymes, protease and amylase, was obtained from A. flavus KU 153, isolatd from South Korea for its high protease production by successive ultra-violet light irradiation, Two glucoamylases from the mutant strain selected were purified from wheat branculture by successive salting out, followed by dialysis and column chromatography, and their characteristics were compared with those of the wild strain. Glucoamylase production of the mutant selected was increased about 3.3 times compared with the wild strain, and 2.1 times compared with the parental strain, ${\alpha}-amylase$ activity of the mutant selected was about 2 times hugher than that of the wild strain or the parental strain. Protease and cellulase productivities of the muant selected were all alike compared with those of the highly proteolytic mutant, the parental strain. Therefore, it was considered that the back mutation on the protease production did not occurred in the formation process of the glucoamylase producing mutant. Total activities of glucoamylase I and II from the mutant selected were 2.86 and 3.65 times higher compared with those from the wild strain, respectively. Considering the optimal pH-thermal stability and Km-Vmax value of glucoamylase I and II from both strains, wild and mutant, it was deduced that the characteristics of glucoamylase I and II from the wild strain did not altered during the mutation process. Therefore, it was concluded that the selected mutant did not induce the formation of another glucoamylase isozyme, or the changes in the characteristics of the glucoamylase, but induce the productivity of the same glucoamylase I and II by the action of regulatory gene.

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