• 제목/요약/키워드: Reversed-phase HPLC

검색결과 358건 처리시간 0.026초

중년 여성에 있어 타우린 보충 급여가 혈청 지질 과산화물 농도에 미치는 영향 (Effects of Taurine Supplementation on Serum Lipidperoxide Levels in Middle-aged Women)

  • 안창순
    • 한국식품영양학회지
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    • 제20권4호
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    • pp.440-449
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    • 2007
  • The purpose of this study was to examine the effects of taurine supplementation on serum lipidperoxide(TBARS), a risk factor for cardiovascular disease. The subjects were 22 healthy middle-aged women(33 to 54 years). Serum lipids, thiobarbituric acid reactive substances(TBARS), and plasma taurine levels were measured before and after supplying 3 g of taurine per day for 4 weeks. Plasma taurine was analyzed by Dabsyl-Cl(4-dimethylamino azobenzen-4-sulfonyl-chloride) derivatization and reversed-phase HPLC. Serum TBARS was measured by the Yagi method. Daily dietary taurine intake was calculated by food frequency questionnaire method. The weight and height means of the 22 subjects were $57.9{\pm}5.2$ kg and $159.2{\pm}5.2$ cm, respectively. Their percent body fat and waist/hip ratio(WHR) were 26.8% and 0.84, respectively, which were slightly higher than the average for middle-aged Korean women. Serum TC, TG and LDL-C levels tended to decrease after taurine supplementation, but HDL-C was not changed. A positive correlation between plasma taurine and HDL-C was shown after taurine supplementation. The serum TBARS concentration was significantly decreased from $5.05{\pm}0.84nmol/d{\ell}$ to $4.17{\pm}0.64nmol/d{\ell}$ after taking taurine(p<0.01), and the plasma taurine concentration was significantly increased from $63.7{\pm}14.2{\mu}mol/{\ell}$ to $73.8{\pm}16.6{\mu}mol/{\ell}$ after taurine supplementation(p<0.05). The average dietary intake of taurine was $178.5{\pm}50.4$ mg/day, which is similar to the average daily taurine intake of Korean women. In conclusion, taurine is an effective nutrient that antagonizes TBARS levels. Therefore, this study suggests that a sufficient taurine intake may be an effective way to prevent cardiovascular disease such as atherosclerosis.

OGS RAAM2000을 이용한 유지방구막 PAS-7 당단백질의 당쇄구조 해석 (Analysis of Sugar Chain Structure of PAS-7 Glycoprotein from Bovine Milk Fat Globule Membrane by US RAAM 2000)

  • 석진석
    • 한국축산식품학회지
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    • 제21권4호
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    • pp.367-373
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    • 2001
  • MFGM 당단백질의 하나인 PAS-7을 GPC 및 affinity chromatography에 의해 정제하여 2-AB로 형광 표식한 후, anion-exchange column 및 reversed-phase column을 이용해 5개의 성분을 분리하였다. 그 중 가장 상대량이 많은 성분 e에 대하여 RAAM2000을 이용한 당쇄구조 해석을 실시하여, 성분 e는 RAAM2000 GPC에 의하여 4개의 성분으로 분리되어 각각 calibration standard 12.10, 8.88, 5.84 및 4.86GU의 용출위치에 검출되었다. 이 용출위치와 당쇄구조는 livrary의 component-7457과 일치하며, 12.2GU의 크기로 분자량은 1788로 판단되며 library의 당쇄와 약 85%의 확률로 일치했다. 그 결과 성분 e의 당쇄구조는 환원말단에 $\alpha$1-6결합된 fucose를 1개 함유하며, core부분의 비환원말단에 N-acetyllactosamine branch를 2개 함유한 전형적인 biantennary 당쇄구조인 것으로 추축되어, 이전 HPLC, acetolysis, sequential exoglycosidase 소화, NMR분석에 의해 보고된 성분 7N1A의 구조와 일치함으로써, OGS RAAM2000을 이용한 PAS-7의 당쇄구조 해석의 유용성이 증명되었다.

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Synthesis, Characterization and In Vitro Evaluation of Triptolide-lysozyme Conjugate for Renal Targeting Delivery of Triptolide

  • Zheng, Qiang;Gong, Tao;Sun, Xun;Zhang, Zhi-Rong
    • Archives of Pharmacal Research
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    • 제29권12호
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    • pp.1164-1170
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    • 2006
  • A triptolide-lysozyme (TP-LZM) conjugate was synthesized to achieve renal specific delivery and to reduce the side effects of triptolide. Triptolide was coupled to lysozyme through succinic via an ester bond with an average coupling degree of 1 mol triptolide per 1 mol lysozyme. The lysozyme can specifically accumulate in the proximal tubular cells of the kidney, making it a potential carrier for targeting drugs to the kidney. The structure of triptolide succinate (TPS) was confirmed by IR, $^{1}H-NMR$, MS and UV. The concentrations of triptolide in various samples were determined by reversed-phase high-performance liquid chromatography (HPLC). In this study, the physicochemical and stability profiles of TP-LZM under various conditions were investgated the stability and releasing profiles of triptolide-lysozyme (TP-LZM) under various conditions. In vitro release trails showed triptolide-lysozyme was relatively stable in plasma (less than 30% of free triptolide released) and could release triptolide quickly in lysosome (more than 80% of free triptolide released) at $37^{\circ}C$ for 24 h. In addition, the biological activities of the conjugate on normal rat kidney proximal tubular cells (NRK52E) were also tested. The conjugate can effectively reduce NO production in the medium of NRK52E induced by lipopolysaccharide (LPS) but with much lower toxicity. These studies suggest the possibility to promote curative effect and reduce its extra-renal toxicity of triptolide by TP-LZM conjugate.

Qualitative and quantitative analysis of furosine in fresh and processed ginsengs

  • Li, Yali;Liu, Xiaoxu;Meng, Lulu;Wang, Yingping
    • Journal of Ginseng Research
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    • 제42권1호
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    • pp.21-26
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    • 2018
  • Background: Furosine (${\varepsilon}$-N-2-furoylmethyl-L-lysine, FML) is an amino acid derivative, which is considered to be an important indicator of the extent of damage (deteriorating the quality of amino acid and proteins due to a blockage of lysine and a decrease in the digestibility of proteins) during the early stages of the Maillard reaction. In addition, FML has been proven to be harmful because it is closely related to a variety of diseases such as diabetes. The qualitative analysis of FML in fresh and processed ginsengs was confirmed using HPLC-MS. Methods: An ion-pair reversed-phase LC method was used for the quantitative analysis of FML in various ginseng samples. Results: The contents of FML in the ginseng samples were 3.35-42.28 g/kg protein. The lowest value was observed in the freshly collected ginseng samples, and the highest value was found in the black ginseng concentrate. Heat treatment and honey addition significantly increased the FML content from 3.35 g/kg protein to 42.28 g/kg protein. Conclusion: These results indicate that FML is a promising indicator to estimate the heat treatment degree and honey addition level during the manufacture of ginseng products. The FML content is also an important parameter to identity the quality of ginseng products. In addition, the generation and regulation of potentially harmful Maillard reaction products-FML in ginseng processing was also investigated, providing a solid theoretical foundation and valuable reference for safe ginseng processing.

Potential Antioxidant Peptides in Rice Wine

  • Rhee, Sook-Jong;Lee, Chung-Yung J.;Kim, Mi-Ryung;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • 제14권4호
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    • pp.715-721
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    • 2004
  • Many food protein hydrolysates have been shown to have antioxidant activities, and recent research focuses on low molecular peptides produced during hydrolysis of food protein. Korean rice wine contains about 60-70% of protein at dry base and originates from raw materials. It has been suggested that the protein is transformed into low molecular weight peptides, and have antioxidant activity during fermentation. The objectives of this study were to evaluate the antioxidant activity of the pre-purified and purified peptides found in Korean rice wine and to identify the responsible peptides. The wine extract of Samhaeju, a traditional Korean rice wine made by low temperature fermentation, was evaporated at $35^{\circ}C$. The two methods employed in the evaluation of antioxidant activity were the DPPH radical scavenging method and the beta-carotene bleaching test. The pre-purified samples showed 808 AAC (Antioxidant Activity Coefficient) and 56.5% AOA (Antioxidant Activity), which were higher than $\alpha$-tocopherol (572 AAC and 78% AOA). The rice wine extract was separated by reversed-phase HPLC. The protective effect of the four most antioxidant active fractions were tested for t-butyl hydroperoxide induced oxidation of healthy human erythrocytes and the byproduct was determined by malondialdehyde formation. Fraction No.5 showed 35% lower MDA concentration as compared to the control. The peptides were further purified using consecutive chromatographic methods and 4 antioxidant peptides were isolated. The amino acid sequences of the peptides were identified as Ile-His-His, Val- Val-His(Asn), Leu-Val-Pro, and Leu(Val)-Lys-Arg-Pro. The AAC value of the synthetic form of the identified peptides was the highest for Ile-His-His.

Radical Scavenging Activity and Content of Cynarin (1,3-dicaffeoylquinic acid) in Artichoke (Cynara scolymus L.)

  • Jun, Neung-Jae;Jang, Ki-Chang;Kim, Seong-Cheol;Moon, Doo-Young;Seong, Ki-Cheol;Kang, Kyung-Hee;Tandang, Leoncia;Kim, Phil-Hoon;Cho, So-Mi K.;Park, Ki-Hun
    • Journal of Applied Biological Chemistry
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    • 제50권4호
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    • pp.244-248
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    • 2007
  • The contents of total phenol and total flavonoid of artichoke (Cynara scolymus L.) were measured. The antioxidant activity of the artichoke was evaluated based on its potential as a scavenging the ABTS radical. These results showed the antioxidant activity of artichoke has a close relationship with the total flavonoid content. The compound showing antioxidant activity was isolated from the artichoke by repeated column chromatography and recrystallization. Based on the spectrometric studies, the compound was identified as 1,3-dicaffeoylquinic acid, known as cynarin. The content of cynarin from heads and leafs of the artichoke determined by $C_{18}$ reversed phase HPLC (high-performance liquid chromatography) coupled with photodiode array detector was 10.15 and 0.67 mg/g, respectively. This compound showed potent antioxidant activities against DPPH and ABTS radicals ($EC_{50}$ = 14.09 and 28.85 ${\mu}M$, respectively).

엽록체내 Protoporphyrin IX(PPIX) 생합성계에서 S-23142의 작용기구 (Action Mechanism of S-23142 on the PPIX Biosynthesis System of Spinacia oleracea L. Chloroplast)

  • 김태준;김진석;조광연;요시다 시게오
    • 한국잡초학회지
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    • 제14권1호
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    • pp.1-7
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    • 1994
  • In vitro 조건에서 protoporphyrin IX(PPIX) 생합성에 미치는 제초제 S-23142와 acifluorfen의 영향을 알아보고자 시금치잎에서 엽록체내의 stroma와 membrane을 분리, 분획하고 형 광검출기가 장착된 역상HPLC를 이용하여 생합성된 PPIX 함량용 분석한 결과는 다음과 같다. 1. PPIX의 생합성이 이루어지는 부위는 엽록체 내의 stroma분획이었고 이는 모두 ALA(${\delta}$-aminolevulinic acid)에 의한 반응산물이었다. 2. PPIX의 생합성에 관련된 in vitro실험은 stroma분획을 이용하는 것이 가장 효율적이었다. 3. In vitro계에서 S-23142와 acifluorfen은 동일하게 PPIX의 생합성을 억제하였으며, 그 작용부위는 엽록체내의 stroma일 것으로 판단되었다.

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Simultaneous Determination of Amitraz, Bromopropylate, Coumaphos, Cymiazole and 2,4-Dimethylaniline in Korean Honey Samples by High-Performance Liquid Chromatography

  • Ryoo, Jae-Jeong;Kim, Se-Hoon;Jeong, Young-Han;Do, Hwan-Suk;Ryu, Jeong-Eun;Kwon, Hae-Young;Jeong, Ji-Yoon;Park, Hye-Jin;Lee, Soon-Ho;Hong, Moo-Ki;Hong, Jong-Ki
    • Bulletin of the Korean Chemical Society
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    • 제29권5호
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    • pp.1043-1047
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    • 2008
  • Simultaneous determination of amitraz, bromopropylate, coumaphos, cymiazole and 2,4-dimethylaniline in 200 honey samples purchased in Korea was performed by reversed-phase high-performance liquid chromatography with multiple UV detection. 2% Acetone in hexane was used for a liquid-liquid extraction and 20-40% water in acetonitrile solutions were used as mobile phases. The LOD for the analytes varied between 0.4 and 1.5 $\mu$g/L and the recoveries were yielded between 64 and 94%. Relative standard deviation of the repeatability of the method is less than 15%. Amitraz was not present in amount above 10 $\mu$ g/L and one for coumaphos and cymiazole and two for bromopropylate, and three for 2,4-dimethylanilne were detected in amount above 10 $\mu$ g/ L. Levels of the acaricide residues found were less than 50 $\mu$ g/L.

국내 시장에서 유통되는 국내·외 초콜릿의 지방산 조성 및 triacylglycerol 조성 분석 (Fatty acid composition and triacylglycerol species of the domestic and foreign chocolates collected from the market)

  • 현진우;신정아;이기택
    • 농업과학연구
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    • 제40권1호
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    • pp.35-45
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    • 2013
  • The fat content, fatty acid composition, trans fatty acid content and triacylglycerol (TAG) composition of 22 chocolates (domestics 8, foreigns 14) collected from the Korea distribution markets were investigated. The crude fat was extracted by acid hydrolysis method and analyzed by gas chromatography (GC) and reversed-phase HPLC for fatty acid and TAG compositions, respectively. The crude fat content of all chocolates varied between 30.11% and 49.59%. The major fatty acids in most of the chocolates were palmitic acid (19.36~31.15 wt%), stearic acid (5.11~36.32 wt%) and oleic acid (18.77~36.68 wt%). Whereas lauric acid (approximately 35.43 wt%) was detected in chocolate fat of sample No. 18. High oleic acid content was observed for the sn-2 position fatty acid with a range from 64.91% to 86.93%. Trans fatty acid contents in domestic chocolates (sample No. 1~8) and foreign chocolates (sample No. 9~22) were 0.03~0.59 wt% (0.01~0.19 g/100g chocolate) and 0.05~6.32 wt% (0.02~1.99 g/100g chocolate), respectively. In TAG composition, TAGs such as POP/PPO(1,3(2)-palmitoyl-2(3)-oleoyl glycerol, PN=48), POS/PSO(palmitoyl-oleoyl-stearoyl glycerol or palmitoyl-stearoyl-oleoyl glycerol, PN=50), SOS/SSO(1,3(2)-stearoyl-2(3)-oleoyl glycerol, PN=50) were mainly detected in most of the chocolates. The peaks of TAG with low PN (ex, 32-34, 36-38, and 40-42) were detected in No. 18 chocolate fat because of containing short chain fatty acid such as lauric acid.

Evidence for Existence of a Water-Extractable Anticoagulant in an Earthworm, Lumbricus rubellus

  • Woo, Jeong-Im;Bahk, Yun-Kyung;Yu, Kyoung-Hee;Paik, Seung-R.;Chang, Chung-Soon
    • BMB Reports
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    • 제29권6호
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    • pp.500-506
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    • 1996
  • We have isolated a water-extracted novel regulator for blood coagulation from an earthworm, Lumbricus rubellus. As a folk remedy, the earthworm has been known to facilitate blood circulation. After complete heat inactivation of endogenous proteases in the earthworm, an anticoagulant(s) was purified through ammonium sulfate fractionation and three consecutive gel permeation chromatography of Sephacryl S-300, Sephadex G-75, and G-150 by measuring activated partial thromboplastin time (APTT) The anticoagulant was further purified to 2,800 fold with a C4 reversed-phase HPLC This activity was stable under heat ($100^{\circ}C$ for 30 min) and acidic conditions (0.4 N HCl). The effects of this partially purified anticoagulant on thrombin were observed with various substrates such as N${\alpha}$-benzoyl-DL-arginine-p-nitroanilide (BApNA), H-D-phenylalanyl-L-pipecoyl-L-arginine-p-nitroanilide (S-2238), N${\alpha}$-p-tosyl-L-arginine methyl ester (TAME), and fibrinogen as a natural substrate. Only TAME hydrolysis, due to an esterase activity of the enzyme, was inhibited among the chromogenic substrates. In addition, the anticoagulant not only inhibited the conversion of fibrinogen to fibrin but also prolonged the fibrin clot formation monitored with the in vitro coagulation test. Based on these observations, we suggest the significance of measuring the ability of antithrombotic drugs to inhibit the esterase activity of thrombin. In this report, it was also shown that the earthworm indeed contained a water-extractable, heat- and acid-stable anticoagulant which could be used as a novel antithrombotic agent.

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