이온교환크로마토그래피에서 라이소자임 분리에 미치는 pH와 온도 영향

고관영 ( Kwan Young Ko ),김인호 ( In Ho Kim ) 한국화학공학회 2014 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.52 No.1

Lysozyme amounts to 0.3% in egg white and functions as an agent of cell lysis and activator of tissue reconstruction. Ion exchange chromatography is the most useful method of separation among affinity chromatography, ion exchange chromatography, and ultra-filtration. The aim of present study is to find the optimum pH and temperature for the separation of lysozyme in egg white within cation exchange gel filled glass column. And we compared results of experiments with those of simulations. Phosphate buffer was used, and pH and temperature were varied as 5~7 and 25~40 oC respectively. RP-HPLC was the tool for the retention time identification and quantitative analysis of lysozyme. OriginPro 8 measured the peak area of lysozyme chromatogram and quantified the eluted lysozyme. Largest amount of lysozyme was separated under the conditions of pH 5 and T 25 oC.

Determination of trace level carbonate ion in Mg–Al layered double hydroxide: Its significance on the anion exchange behaviour

Jui Chakraborty,Swapankumar Ghosh,Somoshree Sengupta,Sudip Dasgupta,Manjusha Chakraborty,Sudipta Mallik,Kamal Lal Das,Debabrata Basu 한국공업화학회 2012 Journal of Industrial and Engineering Chemistry Vol.18 No.6

The presence of trace level (<1 ppm) carbonate ion in the interlayer space of MgAl-LDH has been estimated by ion chromatography technique using a novel inverse chemical suppression method using 0.5 mM H2SO4 as mobile phase and 25 mM LiCl as regenerating solution. The presence of carbonate ion in the LDH structure was found to affect the anion exchange capacity of MgAl-LDH. This has been illustrated by carrying out anion exchange of nitrate ion in the MgAl-LDH structure with chloride ion. The importance of this simple but accurate technique for the carbonate ion estimation is demonstrated and reported in this communication.

The effect of pH and temperature on lysozyme separation in ion-exchange chromatography

Kwan-Young KO,Hong-Gyu LIM,Il-Song LEE,In-Ho KIM 한국생물공학회 2013 한국생물공학회 학술대회 Vol.2013 No.10

Simultaneous Determination of Anions and Cations in Natural Water by Ion-exclusion/Cation-exchange Chromatography with a Weakly Acidic Cation-exchange Resin Column

Lee, Kwang-Pill,Choi, Seong-Ho,Park, Yu-Chul,Bae, Zun-Ung,Lee, Mu-Sang,Lee, Sang-Hak,Chang, Hye-Yong,Kwon, Se-Mok,Kazuhiko Tanaka Korean Chemical Society 2003 Bulletin of the Korean Chemical Society Vol.24 No.9

The simultaneous determination of anions ($SO_4 ^{2-},\;Cl^-,\;and\;NO_3^-$) and cations ($Na^+,\;NH^{4+},\;K^+,\;Mg^{2+},\;and\;Ca^{2+}$) in natural water obtained by Nakdong River waters system in Korea were performed by ion-exclusion/cationexchange chromatography with conductimetric detection. The stationary phase was a polymethacrylate-based weakly acidic cation-exchange resin column in the $H^+$-form and a weak-acid eluent. When using only a 1.4 mM sulfosalicylic acid/6 mM 18-crown-6 ether as an eluent, good resolution of both anions and cations, minimum time required for the separation, and satisfactory detection sensitivity were obtained in a reasonable time. The method was successfully applied to the simultaneous determination of anions and cations in natural waters.

A sensitive and fast HPLC method for determination of Isosorbide 5-mononitrate in human plasma using ion-pair chromatography: application to bioequivalence study

Olyaei Elmira,Samiei Nasim 한국약제학회 2020 Journal of Pharmaceutical Investigation Vol.50 No.5

Purpose Isosorbide 5-mononitrate (5-ISMN) is a charged, hydrophilic, and low ultraviolet-absorbing molecule which is challenging to determine specifically and sensitively in biological matrices. A sensitive and fast ion-exchange chromatography method for determining 5-ISMN in plasma has been developed. Methods The plasma was deproteinized by one-step clean up procedure using sodium chloride and methanol. Then supernatant was injected into a Chromolith C18 column (4 μm, 3.9 × 150 mm). The mobile phase was made of acetate buffer (30 mM, pH: 4.7): methanol, 77:23 (v/v) and 1 mM tetrabutylammonium hydroxide with a flow rate of 1.2 mL/min. Ultraviolet detection was done at 225 nm. The established method was fully validated and successfully applied for pharmacokinetic and bioequivalence study of two different sustained release formulations of 5-ISMN as Elantan LA (reference product) and Mononitral (test product) in twelve healthy male volunteers. A single dose, randomized, two-period, and crossover study was performed under fasting condition. Results Isosorbide dinitrate was used as internal standard. 5-ISMN and internal standard were eluted at 2.6 and 3.8 min, respectively. Calibration curve was linear within the concentration range of 10–800 ng/mL for 5-ISMN in plasma. The results revealed that the precision of the method ranged from 2 to 5 and relative errors lied within 0.43–4.2% regardless of whether the analysis was performed intraday or interday. By this method, the limits of quantitation and detection of 10 ng/ mL and 5 ng /mL were achieved by a sample size of 430 μL plasma. Main pharmacokinetic parameters for test and reference products were 494 ± 210.2 and 500 ± 202.5 for Cmax and 6359 ± 1235 and 6995 ± 1371 for AUC 0-36, respectively. The 90% confidence intervals around ratios (test/reference) from logarithmic transformed exposure values were 99–110 for Cmax and 88–105 for AUC 0–36. Conclusion It can be concluded that both products are bioequivalent. In vitro dissolution test was also carried out together with bioequivalence study and in vitro/in vivo correlation was successfully established.

Purification of Therapeutic Serums of Snake Anti-Venom with Caprylic Acid

Norouznejad, Nilofar,Zolfagharian, Hossein,Babaie, Mahdi,Ghobeh, Maryam KOREAN PHARMACOPUNCTURE INSTITUTE 2022 Journal of pharmacopuncture Vol.25 No.2

Objectives: Antivenom serums have been used extensively for over a century and are the only effective treatment option for snake bites and other dangerous animal envenomations. In therapeutic serum centers, a wide range of antivenoms is made from animal serum, mainly equine and sheep, that are immunized with single or multiple venoms. This work aimed to use caprylic acid (CA) to purify therapeutic snake antivenom. Methods: Plasma was obtained from equine immunized with a mixture of venoms. Immunized plasma was obtained by precipitation of different concentrations (2-5%) of CA. This methodology was compared to that based on ammonium sulfate (AS) precipitation. Sediment plasma proteins were purified by ion-exchange chromatography. Protein assay, SDSPAGE, and agar gel diffusion were performed. Results: The total protein precipitation with AS was higher than precipitation with CA, but the best results were obtained when CA was added to the plasma until a final CA concentration of 5% was reached. Chromatography and electrophoresis indicated a stronger band for the 5% CA, and the gel diffusion assay showed antigen-antibody interaction in the purified serum. Conclusion: The use of CA compared to the routine method for purifying hyperimmune serums is a practical and cost-effective method for preparing and producing therapeutic serums. It constitutes a potentially valuable technology for alleviating the critical shortage of antivenom in Iran.

산수유(山茱萸) 종자(種子)의 독성(毒性)과 렉틴 성분(成分)

정시련,전경희,박소영,장순자,Chung, See-Ryun,Jeune, Kyung-Hee,Park, So-Young,Jang, Soon-Ja 한국생약학회 1993 생약학회지 Vol.24 No.2

The pericarp of Cornus officinalis is well known famous medicinal drug in oriental countries. In this work, we have tried to evaluate the toxicity and also to find the lectin components from this seed. The lyophilized seed extract was lethal to experimental mouse at $250{\sim}300mg/kg$ and this toxic components were related to proteins. The lectins components were partially purified from the extract by ion exchange column chromatography. These lectins were relatively stable at temperature variations and also stable at pH $4{\sim}7$. The activity of these lectins did not inhibit by common carbohydrates molecules.

Purification and Characterization of Bacterial Aryl Alcohol Oxidase from Sphingobacterium sp. ATM and Its Uses in Textile Dye Decolorization

Dhawal P. Tamboli,Amar A. Telke,Vishal V. Dawkar,Shekhar B. Jadhav,Sanjay P. Govindwar 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.4

Aryl alcohol oxidase (AAO) produced by dye decolorizing bacteria Sphingobacterium sp. ATM, was purified 22.63 fold to a specific activity of 21.75 μmol/min/mg protein using anion exchange and size exclusion chromatography. The molecular weight of the purified AAO was found to be 71 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE),and confirmed by zymography of AAO using L-dopa. The enzyme showed substrate specificity towards veratryl alcohol, followed by n-propanol. The optimum pH and temperature of purified AAO were found to be 3.0 and 40°C, respectively. The K_m and V_(max) of AAO was 1.1615mM and 3.13 mM/min when veratryl alcohol was used as substrate. Sodium azide showed maximum inhibition while ethylenediamine tetra acetic acid (EDTA), L-cysteine and dithiothreitol showed slight inhibition. Metal ions also showed slight inhibition. HPLC analysis confirmed the degradation of Direct Red 5B. The metabolite obtained after decolorization of Direct Red 5B was characterized as 3 diazenyl 7 [-(phenyl carbonyl) amino] naphthalene-2-sulfonic acid using GC-MS analysis.

山茱萸 種子의 毒性과 렉틴 成分

鄭時鍊,全瓊姬,朴昭映,章順子 영남대학교 약품개발연구소 1993 영남대학교 약품개발연구소 연구업적집 Vol.3 No.-

The pericarp of Cornus officinalis is well known famous medicinal drug in oriental countries. In this work, we have tried to evaluate the toxicity and also to find the lectin components from this seed. The lyophilized seed extract was lethal to experimental mouse at 250~300 mg/kg and this toxic components were related to proteins. The lectins components were partially purified from the extract by ion exchange column chromatography. These lectins were relatively stable at temperature variations and also stable at pH 4~7. The activity of these lectins did not inhibit by common carbohydrates molecules.

Separation for the Determination of 59/63Ni in Radioactive Wastes

Chang Heon Lee,Kie Chul Jung,Kwang Soon Choi,Kwang Youg Jee,won Ho Kim 한국방사성폐기물학회 2005 방사성폐기물학회지 Vol.3 No.4

방사성 폐기물 핵종 재고량 평가에 필요한 핵종분석을 위하여 다양한 매질의 방사성 폐기물 시료로부터 및 의 분리에 관한 연구가 수행되고 있다. Ni은 음이온교환 수지와 Sr-Spec 추출 크로마토그래피 수지로 Re(의 대용물), Nb, Fe 및 Sr을 차례로 분리하는 과정에서 Ca, Mg, Al, Cr, Ti, Mn, Ce, Na, K 및 Cu와 함께 회수되었다. 본 연구에서는 Ni의 선택적 분리기술을 확립하기 위하여 Ni-Spec 추출 크로마토그래피 및 양이온교환수지법으로 이들의 분리거동을 비교하였다. 또한 Ni의 정제와 기체비례계수법으로 방사능을 측정하기에 적합한 계측시료 준비를 위하여 ammonium 및 tartaric 에서 dimethylglyoxime(DMG)에 의한 Ni의 침전거동을 조사하였다 원자력발전소로부터 채취한 폐이온교환수지 시료 용해용액의 화학조성을 모사하여 만든 모의 폐이온교환수지 용액을 사용하여 Re, Nb, Fe 및 Sr 분리과정을 거쳐 최종적으로 분리한 Ni의 회수율은 이었다. 또한 tartaric 에서 DMG에 의한 Ni의 회수율은 이었다. A study on the separation of in various radioactive wastes discharged from nuclear power plants has been performed for a use in their quantification which is indispensible for the evaluation of the radionuclide inventory Ni was recovered along with Ca, Mg, Al, Cr, Ti, Mn, Ce, Na, K, and Cu through the sequential separation procedure of Re(as a surrogate of ), Nb, Fe and Sr by anion exchange and Sr-Spec extraction chromatography. In this research, chemical separation of Ni from the co-existing elements was investigated by cation exchange and Ni-Spec extraction chromatography. Precipitation behaviour of Ni and the co-existing elements with dimethylglyoxime(DMG) was investigated in ammonium and tartaric in order to purify separated Ni fractions and to prepare source for the radioactivity measurement using a gas proportional counter. Recovery of Ni separated through ion exchange chromatographic separation procedure was with relative standard deviation of . In addition, recovery of Ni with DMG in the tartaric was with relative standard deviation of .