• 한국식품과학회지
• /
• 제31권5호
• /
• pp.1378-1385
• /
• 1999

멸치액젓으로부터 항산화활성을 가지는 물질을 분리하기 위하여 용매추출 및 chromatography를 행하였다. 모든 용매분획들에 항산화활성이 있었으며 그 중 BuOH층과 수용액층이 소금을 함유하고 있었으나 항산화활성이 우수하였다. 수용액층을 투석막(MWCO 1000)으로 증류수에 대하여 투석하였을 때 투석막 내액의 항산화활성이 소실되었으므로 멸치액젓의 항산화성 물질은 분자량 1000이하의 분자량을 가진 것으로 추정하였다. Sephadex G-50 gel filtration chromatography에서 얻은 항산화물질 분획(tube No. 230-245)로 DE-52 anion exchange chromatography를 행한 결과 column에 결합하지 않고 흘러나온 분획(0 N NaCl용액) 및 0.05N NaCl용액으로 용출된 분획에서 항산화활성이 있었으나 $15\;{\mu}M$의 $Fe^{+++}\;ion$의 존재하에서 0 N NaCl 분획만 항산화활성이 유지되었다. Biogel P2 chromatography를 행한 결과 3번과 5번 분획의 항산화 활성이 특히 우수하였으며 TLC확인결과 아미노기가 풍부한 분자들임을 알 수 있었고 아미노산 조성을 분석하였을 때 다른 분획들보다 3번과 5번 분획에 소수성 아미노산이 많이 함유되어 있었다. TLC로 분리하여 항산화활성이 우수한 하나의 band를 HPLC의 C18 column으로 분석하였을 때 3개의 peak 중 항산화활성을 갖은 물질은 methionine 유도체이었으며 이는 멸치젓 수용액층의 많은 항산화물질 중 하나로 여러 아미노산 또는 oligopeptide의 유도체들의 상승효과가 멸치액젓의 항산화성을 유지하는 것으로 사료된다.

• 한국식품과학회지
• /
• 제7권2호
• /
• pp.74-81
• /
• 1975

생(生)배추, $6^{\circ}{\sim}7^{\circ}C$ 숙성(熟成)김치, $22^{\circ}{\sim}23^{\circ}C$ 숙성(熟成)김치를 ion-exchange chromatography로 총유기산(總有機酸)을 분리(分離)하고, silicic acid partition column chromatography로 각유기산(各有機酸)으로 분리(分離), 정량(定量)한 후(後) paper chromatography로 정성확인(定性確認)하였다. 생(生)배추, $6^{\circ}{\sim}7^{\circ}C$ 숙성(熟成)김치, $22^{\circ}{\sim}23^{\circ}C$ 숙성(熟成)김치에는 maleic, fumaric, lactic, succinic, malonic, oxalic, glycolic, malic, citric, tartaric acid 의 유기산(有機酸)과 $H_2SO_4$, $H_3PO_4$ 의 무기산(無機酸)이 들어 있다. 생(生)배추에는 유기산(有機酸)이 염(鹽)의 형태(形態)로 많이 들어 있으며, malic acid가 가장 많고, $6^{\circ}{\sim}7^{\circ}C$ 숙성(熟成)김치, $22^{\circ}{\sim}23^{\circ}C$ 숙성(熟成)김치에는 lactic acid+succinic acid 가 가장 많다. $6^{\circ}{\sim}7^{\circ}C$ 숙성(熟成)김치에는 $22^{\circ}{\sim}23^{\circ}C$ 숙성(熟成)김치보다 lactic acid와 succinic acid의 생성량(生成量)이 많으며, citric acid의 생성량(生成量)은 비슷하고, oxalic, malic, tartaric, fumaric, malonic, maleic, glycolic acid 의 생성량(生成量)은 적다. $6^{\circ}{\sim}7^{\circ}C$ 숙성(熟成)김치가 $22^{\circ}{\sim}23^{\circ}C$ 숙성(熟成)김치보다 더 상큼한 산미(酸味)가 많고 맛이 좋으며 이는 많이 생성(生成)된 lactic acid와 succinic acid 가 관련(關聯)이 되는 것 같다.

• 한국동물학회지
• /
• 제7권1호
• /
• pp.1-5
• /
• 1964

Triglycin 수용액에 미치는 gamma 선의 영향을 low voltage electrophoresis, paper chromatography 및 carbonyl 화합물정량의 세 방법으로 연구하여 다음 결과를 얻었다. 1. 조사된 triglycine 분해산물중 한 성분이 동 ion 과 산화합물을 이루었으며 triglycine에 비하여 높은 음전성을 나타내었다. 2. 조사된 triglycine 분해산물중 다섯 성분이 paper chromatography 에 의해 분리되었으며 이중 둘이 동정되었다. 3. 조사된 triglycine 으로부터 생성되는 carbonly 화합물의 양은 $5{\times}10^{20}\;ev/ml$ 까지는 증가하였으며 이로부터 $1.9{\times}10^{21}\;ev/ml$ 까지의 고선량에서는 오히려 감소를 나타내어 complex system내에 있어서 이차반응에 관여함을 알 수 있었다. 4. 조사된 triglycine에서 glycine을 분리함으로써 peptide bond 의 가수분해의 가능성을 보여주었다.

• Archives of Pharmacal Research
• /
• 제21권3호
• /
• pp.291-297
• /
• 1998

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

• BMB Reports
• /
• 제29권5호
• /
• pp.455-461
• /
• 1996

A serine proteinase was purified from Acanthamoeba culbertsoni by 41~80% ammonium sulfate fractionation, ion exchange chromatography, affinity chromatography and gel filtration chromatography. The molecular weight of the purified enzyme was estimated to be 108.0 kDa by gel filtration chromatography and 54.0 kDa by SDS-PAGE. Therefore, the purified enzyme seemed to be a dimer. Isoelectric point was 4.5. The enzyme activity was highly inhibited by the serine proteinase inhibitors diisopropyl fluorophosphate (OFP) and phenylmethyl sulfonylfluoride (PMSF). It had a narrow pH optimum of 6.5~7.5 with a maximum at pH 7.0. These data suggested that the purified enzyme was a neutral serine proteinase. Optimal temperature was $37^{\circ}C$. It was stable for at least 16 h at $4^{\circ}C$ and $37^{\circ}C$, but it was rapidly inactivated at $65^{\circ}C$ The activity of the purified enzyme was not influenced significantly by $Mg^{2+}$, $Mn^{2+}$, $Zn^{2+}$ or $Ca^{2+}$. However, the enzyme activity was highly inhibited by $Hg^{2+}$ The enzyme degraded type I collagen and fibronectin, but not BSA, hemoglobin, lysozyme, immunoglobulin A or immunoglobulin G.

• Bulletin of the Korean Chemical Society
• /
• 제26권5호
• /
• pp.729-734
• /
• 2005

A sensitive and selective method for quantitation of sofalcone and its active metabolite in human plasma has been established using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Plasma samples were transferred into 96-well plate using an automated sample handling system and spiked with 10 $\mu$L of 2 $\mu$g/mL $d_3$-sofalcone and $d_3$-sofalcone metabolite solutions (internal standard), respectively. After adding 0.5 mL of acetonitrile to the 96-well plate, the plasma samples were then vortexed for 30 sec. After centrifugation, the supernatant was transferred into another 96-well plate and completely evaporated at 40 ${^{\circ}C}$ under a stream of nitrogen. Dry residues were reconstituted with mobile phase and were injected into a $C_{18}$ reversed-phase column. The limit of quantitation of sofalcone and its metabolite was 2 ng/mL, using a sample volume of 0.2 mL for analysis. The reproducibility of the method was evaluated by analyzing 10 replicates over the concentration range of 2 ng/mL to 1000 ng/mL. The validation experiments of the method have shown that the assay has good precision and accuracy. Sofalcone and its metabolite produced a protonated precursor ion ([M+H]$^+$) of m/z 451 and 453, and a corresponding product ion of m/z 315 and 317, respectively. Internal standard ($d_3$-sofalcone and $d_3$-sofalcone metabolite) produced a protonated precursor ion ([M+H]$^+$) of m/z 454 and 456 and a corresponding product ion of m/z 315 and 317, respectively. The method has been successfully applied to a pharmacokinetic study of sofalcone and its active metabolite in human plasma.

• 분석과학
• /
• 제23권1호
• /
• pp.45-53
• /
• 2010

우라늄 화합물 중에 함유된 염소를 정량하기 위하여 수증기증류 및 열가수분해를 이용한 량 염소의 분리 및 정량법을 개발하였다. 수증기 증류에 의한 시료 중의 염소를 분리하기 위하여 수증기 발생장치, 증류플라스크 및 냉각기 등으로 구성된 장치를 제작 설치하였다. LiCl 표준용액과 모의사용후핵 연료 일정량을 혼합하여 만든 우라늄 화합물 시료 중의 염소를 분리하기 위하여 혼산(0.2 M ferrous ammonium sulfate-0.5M sulfamic acid 3 mL + phosphoric acid 6 mL + sulfuric acid 15 mL)을 이용하여 $140^{\circ}C$로 증류시키고 $90{\pm}5\;mL$를 수집하였다. 열가수분해에 의한 시료 중의 염소를 분리하기 위하여 공기공급장치, 온수공급장치, 석영반응관, 연소로 및 연소보트, 그리고 휘발 염소 흡수장치로 구성된 열가수분해장치를 제작 설치하였다. 일정량의 우라늄 화합물 시료에 반응촉진제($U_3O_8$)를 가하고 1 mL/min의 공기유속과 $80^{\circ}C$의 공급수 온도를 유지하고 $950^{\circ}C$에서 1시간 반응시켜 시료 중의 염소를 분리하였다. 두 방법에 의하여 수집된 각 흡수용액은 일정부피로 희석하고 이온크로마토그래피로 정량하여 회수율을 측정하였다. 금속전환체 잔류 용융염 중의 미량 염소를 이온크로마토그래피로 정량하기 위하여 시료를 공기 및 건조 산화시키고 분쇄한 후 열가수분해하여 염소를 회수하였다.

• Bulletin of the Korean Chemical Society
• /
• 제34권6호
• /
• pp.1745-1750
• /
• 2013

A novel green aqueous mobile phase modified with room temperature ionic liquids (RTILs) was employed in the absence of volatile organic solvents or ion-pairing reagents to analyze thiamine, a very polar compound, by reverse phase high performance liquid chromatography (RP-HPLC). Due to its strongly hydrophilic nature, thiamine was eluted near the column dead time ($t_0$) using a mobile phase without adding RTILs or ion-pairing reagents, even if a 100% aqueous mobile phase, which has weak elution power under reverse phase conditions, was used. Thus, 1-ethyl-3-methyl-imidazolium hexafluorophosphate ([EMIM][$PF_6$]), which has the strongest chaotropic effect, was selected as a mobile phase additive to improve retention and avoid baseline disturbances at $t_0$. Various mobile phase parameters such as cation moiety, chaotropic anion moiety, pH and concentration of RTILs were optimized to determine thiamine at the proper retention time. Method validation was performed to assess linearity, intra- and inter-day accuracy and precision, recovery and repeatability; all results were found to be satisfactory. The developed method was also compared to the current official United States Pharmacopoeia (USP) and Korean Pharmacopoeia (KP) methods using an organic mobile phase containing an ionpairing reagent by means of evaluating various chromatographic parameters such as the capacity factor, theoretical plate number, peak asymmetry and tailing factor. The results indicated that the proposed method exhibited better efficiency of thiamine analysis than the official methods, and it was successfully applied to quantify thiamine in pharmaceutical preparations.

• Journal of mucopolysaccharidosis and rare diseases
• /
• 제3권1호
• /
• pp.20-27
• /
• 2017

Purpose: Mucopolysaccharidosis IV (MPS IV) is a disease characterized by deficient activity of N-acetylgalactosamine-6-sulfatase (GALNS) causing excessive lysosomal storage of keratan sulfate (KS). The identification of the relevant disaccharide units of KS after keratanase II digestion followed by liquid chromatography/tandem mass spectrometry detection (LC-MS/MS) is validated and applicable for the preliminary diagnosis of MPS IV. Methods: A total of 67 urine samples were collected and analyzed from 11 MPS IV patients comprising 10 MPS IVA and one MPS IVB patients, and 56 normal controls. Urinary glycosaminoglycan was first precipitated by the Alcian blue method followed by a digestion of keratanase II. The protonated species of the digested disaccharide products were detected by using multiple reaction monitoring experiment. Results: One particular disaccharide of KS was selected. The transition mass-to-charge (m/z) of the parent ion and its daughter ion after collision was $462.0{\rightarrow}97.0$, whereas the chondrosine used as an internal standard in this assay was m/z $353.9{\rightarrow}73.0$. The results corresponded well with the two-dimensional electrophoresis method. The quantities of urinary KS were significantly raised in confirmed MPS IV patients when comparing with those of normal controls ($170.2{\pm}81.1$ vs. $4.06{\pm}1.92{\mu}g/mL$). Conclusion: The LC-MS/MS method for MPS IVA determination is specific, sensitive, validated, and applicable for urinary KS quantification. This method can be used not only as a first-line biochemistry examination of MPS IVA, but also as an outcome survey after enzyme replacement therapy.

• 한국식품영양과학회지
• /
• 제25권4호
• /
• pp.701-707
• /
• 1996

Aspergillus niger SFN-416으로 부터 생성한 xylanase I를 분리.정제하여 특성을 조사하였다. Aspergillus niger SFN-416의 배양액을 ethanol(70%) 침전, $(NH_4)_2-SO_4$(30~90%) 침전, Sephadex G-100 chromatography 및 DEAE-Sephacel ion chromatography 등의 정제과정을 거친 결과, 10.2배 정제되었고, 정제효소의 최적 활성온도는 $50^{\circ}C$ 였다.최적 pH는 3.5이었고, pH 안정성은 6.0 이상에서 활성이 급격히 감소하였다. 또한 금속이온에 대한 효소의 활성은 대부분 억제를 보였고, 특히 $Hg^{2+}$는 18.5%로 가장 낮은 상대활성을 보였지만, $Fe^{2+}$는 117.0%, $Mn^{2+}$는 129.9%로 오히려 효소활성이 증가되었다. 정제 효소의 분자량은 SDS-PAGE에 의하여 31,000 daltons이 었으며, 유기용매에 대한 활성과 안정성은 10%의 methanol, ethanol, isopropanol 및 1-butanol 대하여 모두 낮은 활성을 나타내어 유기용매에는 안정하지 않는 것으로 생각된다.