• Title/Summary/Keyword: chromatographic removal

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Solvent/Detergent Inactivation and Chromatographic Removal of Human Immunodeficiency Virus During the manufacturing of a High Purity Antihemophilic Factor VIII concentrate

  • Kim, In-Seop;Park, Yong-Woon;Woo, Hang-Sang;Chong E. Chang;Lee, Soungmin
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.187-191
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    • 2000
  • A validation study was conducted to determine the efficacy of solvent/Detergent (S/D) inactivation and Q-Sepharose column chromatographic removal of the human immunodeficiency virus (HIV) during the manufacturing of a high purity antihemopilic factor VIII (GreenMono) from human plasma. S/D treatment using the organic solvent, tri (n-butyl) phosphate, and the detergent, Trition X-100, was a robust and effective step in eliminating HIV-1. The HIV-1 titer was reduced from an initial titer of 8.3 log10 TCID50 to undetectable levels within one minute of S/D treatment, HIV-1 was effectively partitioned form factor VIII during Q-Sepharose column chromatography with the log reduction factor of 4.1 . These results strongly assure the safety of GreenMono From HIV.

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Separation of Radiostrontium from Environmental Sample Using Strontim Selective Chromatographic Resin$(Sr.\;Spec^{TM})$ (스트론튬 선택적이온교환수지$(Sr.\;Spec^{TM})$를 이용한 환경시료중의 방사성 스트론튬의 분리)

  • Hong, Kwang-Hee;Lee, Chang-Woo;Choi, Yong-Ho;Lee, Myung-Ho
    • Journal of Radiation Protection and Research
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    • v.20 no.1
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    • pp.1-7
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    • 1995
  • Strontium selective chromatographic material $(Sr-Spec^{TM})$ was investigated for separation of radiostrontium from environmental soil and water sample. This chromatographic material has great capacity of binding of strontium ion in nitric acid media, and has selectivity to permit the separation of stontium from bulk amount of calcium. But the extraction of strontium was reduced by the other interfering ions such as K and Ba. So, in order to apply this material to the soil sample, prior removal treatment of K and Ba was needed. But the Sr-Spec material could provides simple and effective methods for the separation and removal of radiostrontium from liquid sample.

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Membrane for the Removal of Volatile Organic Compounds from Air (대기중의 휘발성 유기물 제거용 분리막)

  • Deng, S.;Lang, K.;Wang, J.;Tremblay, A.;Matsuura, T.
    • Membrane Journal
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    • v.7 no.1
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    • pp.22-30
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    • 1997
  • Porous asymmetric membranes were prepared from polyetherimide polymer by the phase-inversion technique under different conditions. The performance of the membranes was tested for the removal of acetone vapour from nitrogen. A membrane which showed a high acetone permeability and a high selectivity was chosen and tested further for the separation of different organic vapours from nitrogen. The molecular structure of organic vapours and the selectivity were correlated. A strong correlation was also found between the chromatographic retention time of the organic vapour and the selectivity. These experimental results led to the conclusion that the sorption is the factor governing the separation of volatile organic compounds from nitrogen. A membrane was also prepared by coating the surface of a porous polyetherimide membrane with silicone rubber. The performance of membranes with and without silicone rubber coating was compared.

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Chemical coagulation and sonolysis for total aromatic amines removal from anaerobically pre-treated textile wastewater: A comparative study

  • Verma, Akshaya K.;Bhunia, Puspendu;Dash, Rajesh R.
    • Advances in environmental research
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    • v.3 no.4
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    • pp.293-306
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    • 2014
  • The present study primarily focuses on the evaluation of the comparative effect of chemical coagulation and ultrasonication for elimination of aromatic amines (AAs) present in anaerobically pretreated textile wastewater containing different types of dyes including azo dyes. Color and COD reduction was also monitored at the optimized conditions. The production of AAs was measured spectrophotometrically in the form of total aromatic amines (TAAs) and also verified with high performance liquid chromatography (HPLC) selectively. A composite coagulant, magnesium chloride (MC) aided with aluminium chlorohydrate (ACH) in an equal ratio (MC + ACH) was utilized during the coagulation process, which yielded 31% of TAAs removal along with 85% of color and 52% of COD reduction. At optimized power (200 W) and sonication time (5 h), an appreciable TAAs degradation efficiency (85%) was observed along with 51% color reduction and 62% COD removal using ultrasonication. The chromatographic data indicate that sulphanilic acid and benzidine types of aromatic amines were produced after the reductive cleavage of utilized textile dyes, which were effectively mineralized after ultrasonication. The degradation followed the first order kinetics with a correlation coefficient ($R^2$) of 0.89 and a first-order kinetic constant (k) of $0.0073min^{-1}$.

Purification and Characterization of Recombinant Human Follicle Stimulating Hormone Produced by Chinese Hamster Ovary Cells

  • NA KYU HEUM;KIM SEUNG CHUL;SEO KWANG SEOK;LEE SUNG HEE;KIM WON BAE;LEE KANG CHOON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.395-402
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    • 2005
  • Biologically active recombinant human follicle stimulating hormone (rhFSH) was produced in Chinese hamster ovary cells and purified by a series of chromatographic steps. The chromatographic steps included anion-exchange chromatography (DEAE Sepharose F/F, Q Sepharose F/F), hydrophobic interaction chromatography (Source 15 PHE), and hydroxyapatite chromatography (Macro-Prep ceramic hydroxyapatite type I). A distinctive step of the purification process developed was the use of ZnCl$_2$ for the removal of non-glycosylated or lowly-glycosylated FSH and impurities through co-precipitation with Zn$^{2+}$. Purified rhFSH was identified and characterized by several physicochemical and biological methods such as gel electrophoresis, high-performance liquid chromatography, amino acid analysis, carbohydrate analysis, and biological activity. The overall yield of the purification was ~$30\%$. The rhFSH preparation obtained showed high purity (>$99\%$) and high in vivo potency (>16,000 IU/mg). Carbohydrate analysis suggested that the purified rhFSH contained approximately $40\%$ (w/w) carbohydrate with di­or tri-antennary structure on average, which is somewhat more heavily sialylated than commercially available rhFSH. In conclusion, the results of these analyses established an identity of the purified rhFSH with natural FSH from human pituitary glands, and furthermore, the purified rhFSH preparation showed higher in vivo potency and was slightly more heavily sialylated than commercially available rhFSH.

Kinetic studies using a linear regression analysis for a sorption phenomenon of 17a-methyltestosterone by Salvinia cucullata in an active plant reactor

  • Adnan, Fahrizal;Thanasupsin, Sudtida Pliankarom
    • Environmental Engineering Research
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    • v.21 no.4
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    • pp.384-392
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    • 2016
  • The aim of this study was to investigate the removal efficiency of $17{\alpha}$-methyltestosterone (MT) from aqueous solution by Salvinia cucullata Roxb. ex Bory in an active plant-based reactor with a specific focus on linear regression analysis for the sorption phenomena of MT onto the plant roots. A high performance liquid chromatographic method using UV detection (245 nm) was used to analyse the samples. The batch experiments of the active plant reactor (APR) were established to investigate the ability of Salvinia cucullata to remove MT from the liquid phase. The results revealed that 40% and 60% removal of MT from the liquid phase was observed at 5 min. and at 4 h, respectively. Salvinia cucullata can effectively remove MT from the aqueous solution in APRs. Kinetic studies revealed that the sorption phenomena of MT by Salvinia is best described using a linearized pseudo - second order model. Based on the kinetic parameters, it is likely that during the first 4 h of the contact (t = 0 to t = 4 h) sorption is the major driving mechanism of the disappearance of MT from aqueous solutions. However, at higher MT concentrations, diffusivity of MT has a significant effect on the migration of MT from the bulk stream to the root surface. The isotherm analysis revealed that the sorption kinetics favourably followed pseudo second-order. The results of isotherm analysis have indicated that the sorption of MT onto the root surfaces of Salvinia cucullata was favourable and almost irreversible.

Effect of HPLC Analytical Procedure upon Determining Drug Content in PLGA Microspheres

  • Heo, Sun-Ju;Lee, Hong-Hwa;Lee, Min-Jung;Sah, Hong-Kee
    • Journal of Pharmaceutical Investigation
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    • v.40 no.3
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    • pp.193-200
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    • 2010
  • The objective of this study was to investigate the effects of sample preparation, HPLC conditions and peak measurement methods upon determining progesterone content of poly-d,l-lactide-co-glycolide microspheres. A series of the microspheres with different formulations was first prepared. To determine their actual drug contents, the microspheres were dissolved in tetrahydrofuran and diluted with various amounts of methanol to precipitate the polymer. After removal of polymeric precipitates, the filtrates were subject to HPLC analysis under versatile experimental conditions. Interestingly, the composition of a sample solution (e.g., the ratio of methanol to tetrahydrofuran) affected the magnitudes of both peak fronting and peak broadening of progesterone. Its peak became broader and more asymmetrical at lower methanol:tetrahydrofuran ratios. Furthermore, its peak height was influenced by the proportion of tetrahydrofuran in a sample solution. Such problems encountered with tetrahydrofuran were exacerbated when a larger volume of the sample solution was injected onto an analytical column. Under our experimental conditions a peak area measurement provided more accurate and reliable determination of progesterone content in various microspheres than a peak height determination. Optimizing the composition of a sample solution, HPLC chromatographic conditions and peak analysis methods was a prerequisite to an accurate determination of progesterone encapsulated within microspheres.

Purification and Properties of Wheat Fructan Exohydrolase (밀 Frucdtan Exohydrolase의 분리 및 특성)

  • Byeong Ryong, Jeong;Thomas L, Housley
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.4
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    • pp.456-464
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    • 1996
  • Fructans are the major vegetative storage carbohydrate in wheat(Triticum aestivum L.). The depolymerization of fructans occurs by the sequential removal of terminal fructosyl residues by a specific fructan exohydrolase(FEH). The objective of this study was to isolate and characterize this enzyme in wheat. From stems and sheaths of field-grown wheat(cv. Clark), FEH was purified 356-fold using salt precipitation and a series of chromatographic procedures including size exclusion, anion exchange, and affinity chromatography. FEH had a molecular weight of 63.7 kD and an optima at pH 5.5 and 3$0^{\circ}C$. The $K_{m}$s for $\beta$(2 longrightarrow1) linked oligofructans varied, from 10 to 37mM, with the lowest $K_{m}$ for tetrasaccharide. The $V_{max}$ increased as degree of polymerization (DP) increased. Wheat FEH hydrolyzed only, $\beta$(2 longrightarrow1) linked fructans but not, $\beta$(2 longrightarrow6) linked timothy fructan or sucrose. The role of this FEH in fructan metabolism in wheat is discussed.sed.

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Isolation of Sorbic Acid from the Mountain Ash Berries (I) (국산(國産) 건조(乾燥)마가목 열매로 부터 Sorbic Acid의 분리(分離)에 관한 연구(硏究) (I))

  • Lee, S.M.;Kim, C.E.;Joe, Y.I.;Tahk, H.K.
    • Korean Journal of Food Science and Technology
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    • v.4 no.1
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    • pp.1-5
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    • 1972
  • This investigation was primarily carried out to establish the optimum conditions for the separation of sorbic acid, as an enzyme inhibitor, from dry mountain ash berries. The berries were crushed to $35{\sim}40\;mesh$, and leached with water to produce a reddish juice, which was concentrated to syrup after evaporation. Parasorbic acid was separated from the syrup by steam distillation in the presence of acid. In this study, the optimum experimental results for separation of parasorbic acid were obtained as follows : (1) the most applicable leaching time of the dry ash berries was about 6 hours at room temperature and less than 4 hours at $40^{\circ}C$ and (2) in steam distillation, after removal of malic acid, addition of 30 ml of sulfuric acid per 50 ml of syrup extracted from 100 g of the ash berries was very adequate. The purity of sorbic acid obtained experimentally through the isomerization technique was qualitatively examined by spectrophotometeric, paper chromatographic, and melting point measurements.

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Sphingolipid Metabolic Changes during Chiral C2-Ceramides Induced Apoptosis in Human Leukemia Cells

  • Baek, Mi-Young;Yoo, Hwan-Soo;Kazuyasu Nakaya;Moon, Dong-Cheul;Lee, Yong-Moon
    • Archives of Pharmacal Research
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    • v.24 no.2
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    • pp.144-149
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    • 2001
  • N-acetylsphingosine (C2-ceramide) is a synthetic water-soluble ceramide mimicking the activity of natural ceramides. By fixing chiral conformation on carbon numbers 2 and 3 in the ceramide structure, four chiral C2-ceramides naming d-erythro-, I-erythro-, d-threo-and 1-three C2-ceramide were synthesized. We have investigated the chiral effects of these C2-ceramides on the sphingolipid metabolism, particularly on both the sphingolipid bio- synthetic pathway and on the degradation pathway. In both HL-60 and U937 cells, the chiral C2-ceramide ($10{\mu}\textrm{m}$) showed sphingosine accumulation monitored fluoromatrically by a high performance liquid chromatographic separation of the sphingoid bases. Most importantly, in HL-60 cells, l-erythro C2-ceramide induced a 50 fold increase in sphingosine as compared to the control, while l-threo C2-ceramide exhibited a minimal 7-fold in-crease. In contrast, sphinganine, another sphingoid base, showed less accumulation by any chiral C2-ceramide tested under the same conditions. These results suggested that chiral C2-ceramide primarilyacts on the sphingolipid degradation pathway rather than on the sphingolipid biosynthetic route. The strong $C_0/G_1$ phase arrest in the cell cycle by treatment of I-erythro C2-ceramide indicates that the blockade of the sphingolipid degradation pathway might be concomitantly involved in the dysfunction of the cell cycle. On the other hand, the fact that all chiral C2-ceramides tested failed to inhibit the activity of sphingosine kinase acting on the removal of sphingosine by producing sphingosine-1 -phosphate demonstrates that chiral C2- ceramides may increase sphingosine by activating various ceramidases by which natural ceramides are divided into sphingosine and free fatty acids. However, the precise steps involved in this interaction are still unknown.

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