• Title/Summary/Keyword: HPAEC

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Neutral and Amino Sugars Composition of a Lectin from Maackia fauriei (Maackia fauriei 유래 렉틴의 중성당 및 아미노당 조성)

  • Na, Kwang-Heum;Park, Byung-Tae;Park, Jae-Wan;Han, Kyong-Jin;Park, Hyun-Joo;Kim, Ha-Hyung
    • YAKHAK HOEJI
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    • v.53 no.1
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    • pp.34-40
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    • 2009
  • The glycosylation of therapeutic glycoproteins can affect their efficacy, stability, solubility, and half-life. Analyzing the composition of monosaccharides, such as that of neutral and amino sugars, is the first step for elucidating the structure of glycan attached to glycoproteins. In the present study, neutral and amino sugars of lectin obtained from Maackia fauriei were analyzed using an enzyme-linked lectinsorbent assay (ELLA) and high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Peroxidase-labeled lectins such as concanavalin A, Ricinus communis agglutinin, and soybean agglutinin were used for ELLA, since they specifically bind to the monosaccharide residue most frequently encountered in a glycan. The hydrosylate of lectin was prepared by treatment with trifluoroacetic acid, which resulted in the lectin mainly possessing the N-glycan consisting of 98.1 pmol Fuc, 342.1 pmol GlcN, 51.9 pmol Gal, 678.9 pmol Man, and 330.7 pmol Xyl. The present results demonstrate that ELLA and HPAEC-PAD are very effective methods for rapidly estimating the types and relative amounts of monosaccharides in intact glycoproteins.

Quantification of Inulo-oligosaccharides Using High pH Anion Exchange Chromatography with Pulsed Amperometric Detector (HPAEC-PAD)

  • Kang, Su-Il;Chang, Yung-Jin;Kim, Kyoung-Yun;Kim, Su-Il
    • Journal of Applied Biological Chemistry
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    • v.42 no.4
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    • pp.166-168
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    • 1999
  • Inulo-oligosaccharides (IOS, $F_n$, n=2-6) were purified from enzymatic hydrolysates of water-soluble extract of Jerusalem artichoke tubers. Quantification of inulo-oligosaccharides was done using high pH anion exchange chromatography with pulsed amperometric detector (HPAEC-PAD) at the concentration range of 10-100 mg/L, which was compared with that of fructo-oligosaccharides (FOS, $GF_n$, n=1-7). Peak areas per mg IOS were higher than FOS at the same degree of polymerization (DP). Specific peak areas of IOS increased proportionally as DP increased up to six, in contrast to FOS showing no linearity.

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Thermo-chemical Conversion of Poplar Wood (Populus alba × glandulosa) to Monomeric Sugars by Supercritical Water Treatment (초임계수에 의한 현사시나무의 당화 특성)

  • Choi, Joon-Weon;Lim, Hyun-Jin;Han, Kyu-Sung;Choi, Don-Ha
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.6
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    • pp.44-50
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    • 2006
  • To characterize thermo-chemical feature of su gar conversion of woody biomass poplar wood (Populus alba${\times}$glandulosa ) by sub- and supercritical water was treated for 60s under subcritical (23 MPa, 325 and $350^{\circ}C$) and supercritical (23 MPa, 380, 400, and $425^{\circ}C$) conditions, respectively. Among degradation products undegraded poplar wood solids existed in aqueous products. As the treatment temperature increased, the degradation of poplar wood was enhanced and reached up to 83.1% at $425^{\circ}C$. The monomeric sugars derived from fibers of poplar wood by sub- and supercritical treatment were analyzed by high performance anionic exchange chromatography (HPAEC). Under the subcritical temperature ranges, xylan, main hemicellulose component in poplar wood, was preferentially degraded to xylose, while cellulose degradation started at the transition zone between sub and supercritical conditions and was remarkably accelerated at the supercritical condition. The highest yield of monomeric sugars amounts to ca. 7.3% based on air dried wood weight (MC 10%) at $425^{\circ}C$.

Comparison of Volatile Fatty Acids, Monosaccharide Analysis and Metabolic Profiling in Rumen Fluid According to Feeding Methods (사료 급여 방식에 따른 반추위액 내 휘발성지방산과 단당류 비교 분석과 대사산물 분석)

  • Eom, Jun-Sik;Lee, Shin-Ja;Lee, Yoo-Gyung;Lee, Sung-Sill
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.12
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    • pp.814-824
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    • 2018
  • This study was conducted to investigate VFA, monosaccharides and metabolites in rumen fluid according to feeding methods. Three castrated Hanwoo steers were used to the $3{\times}3$ Latin square design, 10 day for the diet adaptation period. VFA and monosaccharides which were not detected by HPLC and HPAEC however, those were detected by $^1H-NMR$. Among the metabolites measured by $^1H-NMR$ carbohydrate metabolites, pyruvate was detected only in the rumen fluid before feeding and succinate was detected before and after feeding rumen fluid. In amino acid total 9 metabolites were detected. In lipid metabolites, ethylene glycol was significantly higher (P<0.05) in before feeding Con group. In aliphatic acylic metabolite, trimethylamine N-oxide was no significant difference observed compare to Con group. In this study, many metabolites were observed in the rumen fluid by $^1H-NMR$, and it confirmed that rumen metabolic products were changed by feeding methods.

Studies on the Changes of Oligosaccharide Contents in Rehmanniae Radix preparata According to Various Processing methods (포제에 따른 숙지황(熟地黃)의 당(糖) 성분 변화 연구)

  • Choi, Ho-Young;Kwon, Seung-Ro;Kim, Hyo-Geun;Ham, In-Hye;Lee, Jae-Jun;Lee, Je-Hyeon;Hong, Seon-Pyo;Kim, Do-Hoon
    • The Korea Journal of Herbology
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    • v.22 no.4
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    • pp.261-270
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    • 2007
  • Objective : The 5-HMF was not index material suitable to do the quality control of Rehmanniae Radix Preparata. In this study, We estimated the changes of oligosaccharide contents in Rehmanniae Radix Preparata using high-performance anion-exchange chromatography with pulsed amperometric detection(HPAEC-PAD). Methods : The analysis of oligosaccharide was conducted by HPAEC-PAD with Carbopac PA1, $250{\times}4mm$, 5um, and Carbopac PA1 guard column. Column temperature was kept at $30^{\circ}C$. Elution was carried out at 1000 ${\mu}l/min$ with 70mM NaOH and the injection volume was $10{\mu}l$. Each component was detected by PAD. Results : Nine constituents were found from merchandising Rehmanniae Radix Preparata(MR), while seven constituents were found in various processed Rehmanniae Radix Preparata. Not all constituents were defined but stachyose and raffinose were found in all cases. And The most common constituents of Rehmanniae radix was stachyose. In the course of processing, most of stachyose and raffinose were decreased. Stachyose was decreased slowly in the course of processing with rice wine(RR), amomi and rice wine(AR), and crataegi and rice wine(CR). However stachyose was decreased rapidly in the course of processing with fresh rehmannia juice(FR). The method with crataegi and rice wine(CR) showed the smallest decrease of stachyose. And processing method with crataegi and rice wine(CR) showed the most abundant amount for stachyose after the nineth processing. Conclusion : The changes of oligosaccharides in the course of processing were a very important direct barometers to do the quality control and set up a standard of Rehmanniae Radix Preparata.

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Simultaneous Removal of Nitrogen and Phosphorus Leached from Farming Feed by the Marine Bacteria, Bacillus sp. CK-10 and Bacillus CK-13, Isolated from Shrimp Farming Pond (새우양식장에서 분리한 해양세균 Bacillus sp. CK-10과 Bacillus sp. CK-13에 의한 양식사료에 포함된 질소와 인의 동시제거)

  • Chun Jae-Woo;Ma Chae-Woo;Kahng Hyung-Yeel;Oh Kye-Heon
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.136-141
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    • 2005
  • A bench-scale feasibility study was conducted with solid farming feed to evaluate a treatment process for microbiological removal of nitrogen (N) and phosphorus (P). Strains, Bacillus sp. CK-10 and Bacillus sp. CK-13, were originally isolated from water samples of shrimp farming pond. Simultaneous removal of N/P in marine media was monitored in the co-cultures, CK-10 and CK-13. As the results, $400\;{\mu}M\;NH^{+}_4$ and $400\;{\mu}M\;NO^{-}_2$ were eliminated within 12 hours and $NO^{-}_3$ within 36 hours, and $500\;{\mu}M\;PO^{-3}_4$ was completely disappeared within 36 hours from the media. Cultures of CK-10 and CK-13 were applied for removal of N/P leached from shrimp farming fred. HPAEC-PAD system was used to analyze sugars in farming feed, resulting in resolution of various sugars including glucose, galactose, galatosamine, mannose, and fucose. $0.2\%$ (w/v) Pulp densities of the farming feed contained approximately $33.3\;{\mu}M\;NH^{+}_4,\;12.9\;{\mu}M\;NO^{-}_2.\;81.5\;{\mu}M\;NO^{-}_3\;and\;248\;{\mu}M\;PO^{-3}_4$ which could dissolved within 72 hours of leaching in aqueous solution followed by bacterial removal. Complete bacterial removal of N/P was achieved within 84 hours at $0.2\%$ of the feed in co-cultures, whereas single cultures removed to incompletion of N/P during the incubation period. This work demonstrated that test cultures, CK-10 and CK-13 showed effective removal of N/P derived from shrimp farming feed.

Molecular Structure and Gelatinization Properties of Turnip Starch (Brassica rapa L.)

  • Kim, Nam-Hee;Yoo, Sang-Ho
    • Food Science and Biotechnology
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    • v.14 no.4
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    • pp.470-473
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    • 2005
  • Starch was isolated from turnip (Brassica rapa L.), and to elucidate the structure-function relationship its structural and physical properties were characterized. Morphological structure of the starch was analyzed by SEM (Scanning Electron Microscopy). Most of the starch granules were spherical in shape with diameter ranging from 0.5-10mm. Apart from larger granules ($<10\;{\mu}m$) which dominated the population size of turnip starch, significant amount of small ($0.5-2\;{\mu}m$) and mid-size granules (${\sim}\;{\mu}m$) were also detected. It was revealed that presumably, erosion damages occurred due to the attack of amylase-type enzymes on the surface of some granules. Branch chain-length distribution was analyzed by HPAEC (High-Performance Anion-Exchange Chromatography). The chain-length distribution of turnip starch revealed a peak at DP12 with obvious shoulder at DP18-21. The weight-average chain length ($CL_{avg}$) was 16.6, and a large proportion (11.8%) of very short chains (DP6-9) was also observed. The melting properties of starch were determined by DSC (Differential Scanning Calorimetry). The onset temperature ($T_o$) and the enthalpy change (${\Delta}H$) of starch gelatinization were $50.5^{\circ}C$ and 12.5 J/g, respectively. The ${\Delta}H$ of the retrograded turnip starch was 3.5 J/g, which indicates 28.2% of recrystallization. Larger proportion of short chains as well as smaller average chain-length can very well explain relatively lower degree of retrogradation in turnip starch.

Analysis of the Structure and Stability of Erythropoietin by pH and Temperature Changes using Various LC/MS

  • Chang, Seong-Hun;Kim, Hyun-Jung;Kim, Chan-Wha
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2663-2670
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    • 2013
  • The purpose of stability testing is to provide evidence about how the quality of a drug varies with time under the influence of a variety of environmental factors. In this study, erythropoietin (EPO) was analyzed under different pH (pH 3 and pH 9) and temperature ($25^{\circ}C$ and $40^{\circ}C$) conditions according to current Good Manufacturing Practice (cGMP) and International Conference on Harmonisation (ICH) guidelines. The molecular weight difference between intact EPO and deglycosylated EPO was determined by SDS-PAGE, and aggregated forms of EPO under thermal stress and high-pH conditions were investigated by size exclusion chromatography. High pH and high temperature induced increases in dimer and high molecular weight aggregate forms of EPO. UPLC-ESI-TOF-MS was applied to analyze the changed modification sites on EPO. Further, normal-phase high-performance liquid chromatography was performed to identify proposed glycan structures and high pH anion exchange chromatography was carried out to investigate any change in carbohydrate composition. The results demonstrated that there were no changes in modification sites or the glycan structure under severe conditions; however, the number of dimers and aggregates increased at $40^{\circ}C$ and pH 9, respectively.

Saccharification of lignocellulosics by Supercritical Water (초임계수를 이용한 목질바이오매스의 당화 가능성)

  • Choi, Joon-Weon;Lim, Hyun-Jin;Han, Gye-Sung;Choi, Don-Ha
    • 한국신재생에너지학회:학술대회논문집
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    • 2006.11a
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    • pp.525-528
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    • 2006
  • 아임계 및 초임계수에 의한 목질바이오매스의 당화특성을 분석하기 위하여 분해공정 동안 압력을 23MPa(물의 임계압력)로 고정하고 물의 아임계 온도$(325^{\circ}C,\;350^{\circ}C)$와 초임계 온도$(380^{\circ}C,\;400^{\circ}C,\;425^{\circ}C)$에서 현사시나무 목분을 각각 60초 동안 처리하였다. 생성된 현사시나무의 분해산물에는 액상과 고형분의 분해산물이 섞여 있었다. 각 처리조건에 따른 목질바이오매스의 분해율은 온도가 상승함에 따라 증가하였으며 초임계 온도인 $425^{\circ}C$에서 최고 83.1%의 분해율을 나타냈다. 아임계 및 초임계수에 의해서 생성된 단당류는 고성능 음이온 교환 크로바토그래프(HPAEC)를 이용하여 분석하였다. 목질바이오매스의 초임계수 분해과정에서 처리 온도가 높아지면서 단당류 수율은 증가하는 경향을 보였으며, $425^{\circ}C$에서 가장 높은 7.3%의 단당류 수율을 나타내었다. 아임계 온도 범위에서는 현사시나무의 섬유소 성분 중에서 자일란이 우선적으로 분해되어 자일로스의 생성비율이 비교적 높았으며, 처리온도가 높아지면서 셀롤로오스의 분해에 의한 글루코오스 생성율이 급격히 상승하였다. 이렇게 생성된 단당류 성분들은 고온의 반응조건하에서 열분해 반응에 의해서 더욱 분해되어 퓨란계 화합물로 변형되었다.

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