• Title/Summary/Keyword: column chromatography

Search Result 3,267, Processing Time 0.028 seconds

Characterization and Purification of Anti-Complement Polysaccharide from Spirodela polyrhiza

  • Min, Jin-Gi;Lee, Doo-Seog;Park, Jeong-Heum;Heo, Moon-Soo;Kim, Tae-Jin;Shim, Kil-Bo;Cho, Young-Je
    • Preventive Nutrition and Food Science
    • /
    • v.9 no.1
    • /
    • pp.21-28
    • /
    • 2004
  • We purified and characterized a crude polysaccharide from Spirodela polyrhiza with anti-complement activities. The crude polysaccharide fraction (SP-0) which had potential anti-complement activity was extracted in hot water for 4 hrs at 10$0^{\circ}C$. The ethanol-precipitate, the crude polysaccharide traction (SP-1), showed a potent anti-complement activity. Further purification of the crude polysaccharide (SP-1) was carried out by cetavlon, ion exchange chromatography and gel column chromatography. Among cetavlon fractions, SP-4 showed the most potent anti-complement activity. When 100 $\mu\textrm{g}$/mL of SP-4 was incubated with an equal volume of normal human serum (NHS), the TCH$_{50}$ was reduced by about 78%. When the SP-4 fraction was further purified by DEAE-Sepharose (Cl$^{[-10]}$ ), the SP-4IIa, SP-4IIb and SP-4IIc, absorbed fractions, were almost the same as the anti-complement activities of SP-4. SP-4IIc, having the greatest potential activation and the highest yield by ion exchange chromatography, was further purified by gel column chromatography on a Sepharose CL-6B column. Four polysaccharide fractions of SP-4IIc-1, SP-4IIc-2, SP-4IIc-3 and SP-4IIc-4 were obtained, consisting mainly of arabinose, rhamnose, galactose and glucose, with approximate molecular weights of about 305,000, 132,000, 64,000 and 12,000, respectively. Among these subfractions, SP-4IIc-1 had the most potent anti-complement activity. When the SP-4IIc-1 aggregate was applied to a gel column chromatography in 10 mM and 50 mM NaCl solution, the position of the peak fractions shifted to a low molecular weight region, and the molecular weight of SP-4IIc-1 decreased with increased NaCl concentration in the gel column chromatography. It was found that the self-aggregation formed spontaneously in void volume by gel column chromatography using Sepharose CL-6B in water and the self-aggregation significantly affected the anti-complement function.

Fractionation and Physicochemical Characteristics of Caprine Casein by DEAE-Cellulose (DEAE-Cellulose에 의한 산양유 Casein의 분별 및 이화학적 성질에 관한 연구)

  • Im, Dong-Hyeon;Jeon, U-Min;Han, Gyeong-Sik;Kim, Byeong-Cheol;Hwang, Gwang-Yeon;Kim, Se-Heon
    • Journal of Dairy Science and Biotechnology
    • /
    • v.18 no.1
    • /
    • pp.1-8
    • /
    • 2000
  • This experiment was carried out to study fractionation and physicochemical characteristics of caprine casein. Acid caseins obtained from caprine colostral and normal milk were analyzed by chymosin treatment and fractionated by DEAE-cellulose column chromatography with linear gradient and electrophoresis. Protein, fat and lactose of caprine normal milk were 2.70${\pm}$0.27%, 3.82${\pm}$0.51%, and 4.10${\pm}$0.29%, respectively. More non-protein nitrogen(NPN) was released by chymosin treatment from caprine colostral casein than normal casein. The electrophoretic pattern of caprine casein was not similar to that of bovine casein, Caprine normal casein was fractionated by DEAE-cellulose column chromatography with a 0.08${\sim}$0.18 M NaCl linear gradient into 5 pes with the proportion of 5.27%, 26.07%, 25.97%, 30.40% and 12.29%, respectively. In order to identify the pure fraction, the chymosin-treated caprine normal casein was fractionated by DEAE-cellulosecolumn chromatography with a 0.08${\sim}$0.18 M NaCl linear gradient into 6 peaks with the proportion of 17.06%, 9.10%, 17.85%, 20.11%, 27.03% and 8.85%, respectively.

  • PDF

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

  • 박용곤
    • The Korean Journal of Food And Nutrition
    • /
    • v.12 no.5
    • /
    • pp.508-512
    • /
    • 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%.

  • PDF

Purification of the Vacuolar Arginine Transporter from Neurospora crassa (Neurospora crassa로부터 arginine transporter의 순수분리)

  • ;Weiss, R. L.
    • Korean Journal of Microbiology
    • /
    • v.27 no.2
    • /
    • pp.117-123
    • /
    • 1989
  • Radioactive N-$\alpha$-p-nitrobenzoxycarbonyl (NBZ)-L-[2,$3-^{3}$H] arginyl diazomethane was used as an affinity label for the vacuolar arginine transporter in Neurospora crassa. Vacuolar matrix proteins were removed by fracturing the membranes with freeze-thaw method in dry ice/ethanol bath. Vacuolar membrane proteins were then wasged with 500mM NaCl to remove ionically bound derivatives and peripheral membrane proteins from vacuolar membranes. After dissolved in 1% Titon X-100, dissolved vacuolar memvrane proteins were separated with molecular sieve column chromatography, anion and cation exchange chromatographies. The arginine transporter was purified giving the purification factor of 1136.

  • PDF

UHPLC System Shutdown and Reactivation Advice (UHPLC 시스템 종료 및 재가동 시 가이드)

  • Mark Fever;Gemma Lo
    • FOCUS: LIFE SCIENCE
    • /
    • no.1
    • /
    • pp.8.1-8.3
    • /
    • 2024
  • Ultra-high performance liquid chromatography (UHPLC) systems are integral to modern analytical laboratories, necessitating careful maintenance and operation protocols to ensure optimal performance. This document provides comprehensive guidelines for the proper shutdown and reactivation of UHPLC systems to prevent damage and maintain operational efficiency. • Shutdown: Remove the column and replace it with a union to avoid blockages. Flush the system with a compatible solvent mix, clean mobile phase reservoirs to prevent microbial growth, flush the pump with storage solvent, and clean the autosampler, including the needle and injection port. • Reactivation: Inspect the system for wear or damage, gradually reintroduce mobile phases starting with a weak solvent, reinstall the column securely, and perform system checks on baseline stability, pressure consistency, and detector performance. By adhering to these guidelines, laboratories can ensure the longevity and reliability of their UHPLC systems, maintaining high analytical performance and minimizing downtime. These procedures help prevent common issues such as blockages, contamination, and component wear, thereby supporting efficient and accurate analytical operations.

  • PDF

A NUMERICAL STUDY ON THE FLOW CHARACTERISTICS OF GAS CHROMATOGRAPHIC COLUMN (가스 크로마토그래픽 컬럼의 유동특성에 대한 수치적 연구)

  • Kim T.-A.;Kim Youn J.
    • 한국전산유체공학회:학술대회논문집
    • /
    • 2005.10a
    • /
    • pp.21-26
    • /
    • 2005
  • Gas Chromatography (GC) is a wisely technique used for the separation and analysis of liquid and gas sample. Separation of the sample vapors is achieved via their differential migration through a capillary column with an insert carrier gas. The identity and quantity of each vapor in the mixer can be determined from its retention time in the column and a particular property of the gas, such as thermal conductivity, which can be related to the concentration of sample vapor in the carrier gas. Therefore, the flow characteristics in the spiral gas chromatographic column are numerically investigated in this study. Especially, different pressure drop between the front and the rear of GC column with various flow rates is estimated the governing equations are derived from making using of three-dimensional Naver-Stokes equation with incompressible and laminar model due to the nature of low Reynolds number flow. Using a commercial code, FLUENT, the pressure and flow fields in GC column are calculated with various flow rates. The characteristics of thermal cycling which is one of the most important factors affecting the column efficiency and analysis time is also estimated. Furthermore, numerical analyses are also carried out by using commercial code, ANSYS, with various values of power, which is applied to the heating element located at lower GC column.

  • PDF

Coupled Column Chromatography in Chiral Separation of Salmeterol

  • Kim, Kyeong-Ho;Yun, Hyeong-Won;Kim, Hyun-Ju;Park, Hyun-Ji;Choi, Pok-Wha
    • Archives of Pharmacal Research
    • /
    • v.21 no.2
    • /
    • pp.212-216
    • /
    • 1998
  • A coupled achiral-chiral high-performance liquid chromatographic system has been developed for the determination of the enantiomers of salmeterol, S-(+)-salmeterol and R-(-)-salmeterol in urine. THe salmeterol was separated from the interfering components in urine and quantified on the silica column, and the enantiomeric composition was determined on a Sumichiral OA-4700 chiral stationary phase. The two columns were connected by a switching valve equipped with a silica precolumn. The two columns wer connected by a switching valve equipped with a silica precolumn. The precolumn was used to concentrate the salmeterol in the eluent from the achiral column before backflushing onto the chiral phase. The coupled system was validated.

  • PDF

Purification of Inositol Triphosphate Kinase from Bovine Brain (소의 뇌로부터 Inositol Triphosphate Kinase의 정제)

  • Kim, Jung-Hye;Lee, Jae-Tae
    • Journal of Yeungnam Medical Science
    • /
    • v.13 no.1
    • /
    • pp.46-58
    • /
    • 1996
  • Inositol 1,4,5-triphosphate($InsP_3$) is a second messenger for mobilizing intracellular $Ca^{2+}$. It can be dephosphorylated by soluble and particulate forms on $InsP_3$ 5-phosphatase, or phosphorylated to produce inositol 1,3,4,5-tetrakisphosphate($InsP_3$) by $InsP_3$ 3-kinase. These enzymes represent possible targets for the regulation of the $InsP_3/InsP_4$ signal. $InsP_3$ 3-kinase which catalyses th ATP-dependent phosphorylation of $InsP_3$ was purified from bovine brain tissue. All operation were carried out at $4^{\circ}C$. Fresh tissure was homogenized and centrifuged. The supernatant was pooled. Proteins were precipitated from 10% polyethylene glycol, and suspended solution was applied to DEAE cellulose column for chromatography. As the result of above procedure, two isozymes of $InsP_3$ 3-kinase, I and II were obtained. Each isozyme was applied to Matriz green gel, Calmodulin-Affigel 15 column and subsequent phenyl-TSK HPLC column. Specific activites(SA) and fold of puriety were observed at each purification step of chromatography. At DEAE cellulose chromatography, SA were I, 0.6 and II, 4.8 nM/min/mg, and folds were I, 17.2 and II, 16.6. At Matrix green gel chromatography, SA were I, 18 and II, 11 nM/min/mg, folds were I, 62.1 and II, 38.0. At calmodulin-Affigel 15 column chromatography, SA were I, 19 and II, 13 nM/min/mg, folds were I, 65.5 and II, 44.8. Finally $InsP_3$ kinase I and II were purified 3,103-fold and 2,310-fold, and SA were I, 900 and II, 670 nM/min/mg, respectively. SDS-polyacrylamide gel electrophoresis elucidated 3 distinct fractions of Mr of 145,000, 85,000 and 69,500 from isozyme I, and 2 distinct fractions of Mr of 79,000 and 57,000 from isozyme II.

  • PDF

Determination of terbutaline in human plasma by coupled column chromatography (커플드칼럼크로마토그래피에 의한 사람 혈장 중 테르부탈린의 정량)

  • Ko, Mi Young;Jeon, Sang-Seol;Kim, Kyeong Ho
    • Analytical Science and Technology
    • /
    • v.28 no.2
    • /
    • pp.125-131
    • /
    • 2015
  • A method was developed and fully validated for the determination of terbutaline, a β2-receptor agonist, in human plasma. Plasma samples were prepared by solid-phase extraction with Sep-Pak silica, followed by high-performance liquid chromatography (HPLC). The terbutaline was pre-separated from the interfering components in plasma on a Luna C18 (2) column, and terbutaline and salbutamol as an internal standard were resolved and determined on a Luna Silica column. The two columns were connected by a switching valve equipped with silica pre-column. The pre-column was used to concentrate the terbutaline in the eluent from the C18 column before back-flushing onto the silica column with fluorescence detection at an excitation/emission wavelength of 276/306 nm. The method was shown to be specific by testing six different human plasma sources. Linearity was established for a concentration range of 0.4-20.0 ng/mL with a correlation coefficient of 0.9999. The lower limit of quantitation was 0.4 ng/mL with a precision of 10.1% as C.V.%.