• 제목/요약/키워드: HPLC Chromatography

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Taxol의 방사면역측정을 위한 I-125 표지화합물 합성 (Synthesis of I-125 Labelled Compound of Taxol Analogue for Radioimmunoassay)

  • 오옥두;금준섭;이양호;박용석;편웅범;최창운
    • 대한의생명과학회지
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    • 제3권2호
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    • pp.125-130
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    • 1997
  • Taxol은 diterpenoid구조를 가진 항암제로서, 난소암과 유방암에 탁월한 효과를 보이지만 다른 항암제와 마찬가지로 독성을 가지고 있어 약물의 체내 혈중농도를 모니터링하는 것이 필요하다. 약물의 혈중농도를 모니터링하는 방법은 HPLC법, ELISA법, RIA법 등이 있으나, RIA법이 민감도 측면에서 또한 간편하다는 점에서 장점이 있다. 본 연구에서는 I-125표지항원을 이용한 방사면역측정법을 확립하기 위해 먼저 taxol유도체를 합성하였다. 먼저 taxol의 C-13 탄소의 곁가지에 위치한 C-2'부분의 hydroxy기를 succinic anhydride와 반응시켜 2'-hemisuccinyltaxol (I)을 합성 (반응수율:80%)하였다. 또한 tyramine을 $^{125}$I로 표지하고 gel chromatopaphy를 통해 정제된 [$^{125}$I]iodotyramine (II) (반응수율:58%)을 얻었다. (I)과 (II)를 반응시켜 2'-[$^{125}$I]iodotyramine-hemisuccinyltaxol (III) (반응수율:96%)을 얻어 $^{125}$I 표지항원으로 사용하였다. Taxol에 대한 항체를 획득하기 위해서 (I)을 BSA에 접합반응시켜 2'-hemisuccinyltaxol-BSA접합체를 합성하였으며, 이것을 토끼에 면역 주사하여 anti-taxol serum을 얻었다. 이 항체에 대한 역가 검정실험에서 1:20의 희석비에서 B/F(%)가 약 40%를 보였다. 이와 같은 결과는 2'-[$^{125}$I]iodotyramino-hemisuccinyltaxol을 표지항원으로 한 taxol의 방사면역측정 방법으로 혈청내 taxol의 농도측정이 가능함을 제시해 준다.

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Vitamin D3 and Beta-carotene Deficiency is Associated with Risk of Esophageal Squamous Cell Carcinoma - Results of a Case-control Study in China

  • Huang, Gui-Ling;Yang, Lei;Su, Ming;Wang, Shao-Kang;Yin, Hong;Wang, Jia-Sheng;Sun, Gui-Ju
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권2호
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    • pp.819-823
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    • 2014
  • Objective: The aim was to evaluate roles of vitamin D3 (VD3) and beta-carotene (BC) in the development of esophageal squamous cell carcinoma (ESCC) in a high-risk area, Huai'an District, Huai'an City, China. Methods: 100 new ESCC diagnosed cases from 2007 to 2008 and 200 residency- age-, and sex-matched healthy controls were recruited. Data were collected from questionnaires, including a food frequency questionnaire (FFQ) to calculate the BC intake, and reversed phase high-performance liquid chromatography (RP-HPLC) was used to measure the serum concentrations of BC and VD3. Odds ratios (OR) and 95% confidence intervals (CI) were calculated in conditional logistic regression models. Results: The average dietary intake of BC was $3322.9{\mu}g$ (2032.4-5734.3) in the case group and $3626.8{\mu}g$ (1961.9-5827.9) in control group per capita per day with no significant difference by Wilcoxon test (p>0.05). However, the levels of VD3 and BC in the case group were significantly lower than in the control group (p<0.05). The OR values of the highest quartile and the lowest quartile of VD3 and BC in serum samples were both 0.13. Conclusion: Our results add to the evidence that high circulating levels of VD3 and BC are associated with a reduced risk of ESCC in this Chinese population.

Lack of an Apparent Association between Mycotoxin Concentrations in Red Chili Peppers and Incidence of Gallbladder Cancer in India : an Ecological Study

  • Ikoma, Toshikazu;Kapoor, Vinay Kumar;Behari, Anu;Mishra, Kumudesh;Tsuchiya, Yasuo;Asai, Takao;Endoh, Kazuo;Okano, Kiyoshi;Nakamura, Kazutoshi
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권7호
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    • pp.3499-3503
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    • 2016
  • Our recent studies conducted in South America have shown that mycotoxin contamination of red chili peppers (RCPs) may be associated with an increased risk of gallbladder cancer (GBC). Whether this relationship exists in India, a country with a high incidence of GBC and high consumption of RCPs, is unclear. We therefore measured concentrations of aflatoxins (AFs) and ochratoxin A (OTA) in RCPs from areas of low, medium, and high incidence of GBC in India, and compared these concentrations with GBC incidence in each area. Twenty-one RCP samples were collected from nine cities (eight from a low-incidence area, five from a medium-incidence area, and eight from a high-incidence area). Concentrations of AFs and OTA were measured using high-performance liquid chromatography. No significant differences in mean concentrations of AFs and OTA were found in the three areas. AFB1 levels in the low-incidence area ($10.81{\mu}g/kg$) and high-incidence area ($12.00{\mu}g/kg$) were more than 2.2 and 2.4 times higher compared with the maximum permitted level of AFB1 in spices ($5.0{\mu}g/kg$) set by the Commission of the European Communities, or that ($4.4{\mu}g/kg$) obtained in our previous study in Chile. Our results show that the mean concentrations of mycotoxins in RCPs are similar among the three areas in India with different incidences of GBC. Further studies with human subjects are needed to evaluate any association between AFB1 and GBC.

양식 넙치(Paralichthys olivaceus)에 경구투여한 Ampicillin 및 Amoxicillin의 어류체내 잔류량의 변화 (Residues of Ampicillin and Amoxicillin in Olive Flounder Paralichthys olivaceus Following Oral Administration)

  • 손광태;조미라;오은경;목종수;권지영;이태식;송기철;김풍호;이희정
    • 한국수산과학회지
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    • 제44권5호
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    • pp.464-469
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    • 2011
  • Residues of ampicillin and amoxicillin in the muscles of olive flounder Paralichthys olivaceus were investigated after oral doses of 20 mg/kg body weight/day of ampicillin and 40 mg/kg body weight/day of amoxicillin in a first examination and 40 mg/kg body weight/day of ampicillin and 80 mg/kg body weight/day of amoxicillin in a second examination for 5-6 days. The ranges of seawater temperatures in the $1^{st}$ and $2^{nd}$ examinations were $13.4-15.6^{\circ}C$ and $16.8-21.4^{\circ}C$, respectively. Ampicillin and amoxicillin concentrations were analyzed by high performance liquid chromatography (HPLC). The average recovery rates of ampicillin and amoxicillin in fish samples ranged from 84.8-95.2% and 100.8-103.8%, respectively. Residual concentrations in the olive flounder peaked on the $5^{th}$ day, with average concentrations of 0.075 mg/kg and 0.311 mg/kg in the $1^{st}$ examination, and 0.098 mg/kg and 0.630 mg/kg in the $2^{nd}$ examination for ampicillin and amoxicillin, respectively, with maximum concentrations in muscle. In the $1^{st}$ examination, ampicillin concentrations in olive flounder after withdrawals of 1 and 3 days were 0.041 mg/kg and 0.023 mg/kg, respectively, and amoxicillin concentrations after withdrawals of 1 day and 10 days were 0.172 mg/kg and 0.023 mg/kg, respectively. In the $2^{nd}$ examination, the ampicillin concentrations in olive flounder after withdrawals of 1 and 3 days were 0.041 mg/kg and 0.023 mg/kg, respectively, and amoxicillin concentration after withdrawals of 1 day and 10 days were 0.172 mg/kg and 0.023 mg/kg, respectively. We suggest that the recommended withdrawal periods should be 3 days for ampicillin and 10 days for amoxicillin in the olive flounder.

Effect of Solar Irradiances on Growth and Pigmentation of Antarctic Red Algae, Kallymenia antarctica and Palmaria decipiens

  • Han, Tae-Jun;Han, Young-Seok;Lee, Min-Soo;Park, Jin-Hee;Cho, Man-Gi;Koo, Jae-Gun;Kang, Sung-Ho
    • Ocean and Polar Research
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    • 제25권4호
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    • pp.427-435
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    • 2003
  • Growth and pigment responses to different levels of solar radiation with or without ultraviolet (UV)-B component $({\lambda}=280-315nm)$ were investigated in Antarctic rhodophytes, Kallymenia antarctica and Palmaria decipiens, collected around King George Island during the summer of 2000. In K. antarctica specific growth rate, based on thallus area or fresh weight, decreased with increasing solar irradiances while P. decipiens were relatively insensitive to the effects of light. It is noticeable that the presence or absence of UV-B had no significant effect on growth for either species. However, K. antarctica showed a more pronounced reduction in chlorophyll (Chl a) concentrations at higher irradiances in the presence of UV-B. In P. decipiens, Chl a concentrations did not differ despite radiation level fluctuations being lower albeit than initial measurements. Thallus thickness was greater in K. antarctica than in P. decipiens. There were higher relative amounts of UV-absorbing pigments (UVAPs) in P. decipiens than in K, antarctica. The single absorbance peak obtained from the methanol extracts was resolved into three (316,332 and 346nm) in K. antarctica and four peaks (315,326,333 and 349 nm) in Palmaria as a result of the fourth-derivative. After 7 days exposure to solar radiation, the amount of UVAPs in K. antarctica was significantly reduced to a similar degree at all light levels, whereas that of P. decipiens remained unchanged except at 5% of surface irradiance. High performance liquid chromatography (HPLC) analysis of purified extracts indicated that P. decipiens possesses porphyra-334 in addition to three other mycosporine-like anlino acids (MAAs; asterina-330, palythine, shinorine), which are commonly present in K. antarctica. Significantly lower tolerance of K. antarctica to high levels of solar radiation may be connected with its usual absence in the eulittoral, while the active growth and elastic pigment responses of P. decipiens over a wide range of solar irradiance levels up to full sunlight seems to correspond well with its wide vertical distribution from rock pools down to 25-30m.

이중 채널 유리코일 샘플러를 이용한 대기 중 포름알데하이드 측정 (Measurement of Formaldehyde in the Atmosphere using a Dual-channel Glass Coil Sampler)

  • 박승식;홍상범;이재훈;조성용;김승재
    • 한국대기환경학회지
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    • 제22권2호
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    • pp.259-266
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    • 2006
  • A dual-channel glass coil sampling technique was used to measure hourly formaldehyde concentration in the ambient air. The dual-channel coil sampling assembly consists of three parts; an all-pyrex 28-turn coil made of 0.2-cm internal diameter glass tubing for gas-liquid contact and scrubbing of soluble gases, an inlet section upstream of the coil for introducing sample air and scrubbing solution, and a widened glass section downstream of the coil for gas-liquid separation. The scrubbing solution used was a dilute aqueous DNPH (dinitrophenylhydrazine) solution. Hourly concentration of formaldehyde was determined at a Gwangju semi-urban site during two intensive studies between September and October using the dual channel glass-coil/DNPH sampling technique and HPLC (High Performance Liquid Chromatography) analysis. The mean concentration was 1.7($0.4{\sim}4.7$) and 3.0($0.5{\sim}19.1$) ppbv for the September and October intensives, respectively, which are considerably low, compared to those measured in polluted urban areas around the world including several urban areas of Korea. The diurnal variation showed significant increase of formaldehyde in the daytime suggesting the dominance of formation of formaldehyde due to photochemical oxidation of methane and other hydrocarbons. An increase in the formaldehyde sometimes in the night might be due to an increase in primary source, i.e. traffic emissions. It was also found that rapid increase in formaldehyde levels from 3.0 to 19.1 ppbv in the afternoon on October 20 was due to plumes from burning of agricultural wastes such as rice straw and stubble. It is expected from the measurement data that the constructed dual-channel glass coil sampling system can be utilized for measuring atmospheric concentration of the formaldehyde with high time resolution.

카페인산의 효소적 산화반응으로부터 췌장 지방분해효소 저해 물질의 분리 (Secondary Metabolites from Enzymatic Oxidation of Caffeic Acid with Pancreatic Lipase Inhibitory Activity)

  • 김태훈;김명권
    • 한국식품영양과학회지
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    • 제44권12호
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    • pp.1912-1917
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    • 2015
  • 천연식물에 광범위하게 존재하는 대표적인 페닐프로파노이드 화합물인 caffeic acid에 대해 배 유래의 polyphenol oxidase로 산화반응을 수행하여 상대적으로 높은 pancreatic lipase 저해 활성($IC_{50}$; $161.2{\pm}2.8{\mu}g/mL$)을 확인하였으며, 이는 caffeic acid와 비교하였을 경우 활성이 상승함을 알 수 있었다. Caffeic acid 산화반응물에 대해서 $C_{18}$ 겔을 활용한 column chromatography를 수행하여 4종의 리그난 화합물을 분리하였고, 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 phellinsin A(2), caffeicinic acid(3), isocaffeicinic acid(4), 7,8-erythro-caffeicin(5)으로 동정하였다. 이들 화합물중 phellinsin A(2)는 $IC_{50}$ 값이 $66.3{\pm}2.6{\mu}M$로 가장 강한 효능을 나타내었으며, 다음으로 caffeic acid 2분자의 산화 결합을 통해 생합성된 caffeicinic acid(3)의 $IC_{50}$ 값이 $109.6{\pm}3.7{\mu}M$의 저해능을 나타내었다. 배에 존재하는 polyphenol 산화효소에 의해 생합성된 caffeic acid 이량체가 pancreatic lipase 저해 활성 물질임을 확인하였으며, 이들 활성은 caffeic acid가 결합 양상에 따른 화합물의 구조에 따라 다름이 시사되었다. 향후 이들 활성물질의 활성 기작에 대한 연구가 필요하며 본 연구 결과는 보다 우수한 pancreatic lipase 저해능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을 뿐만 아니라 항비만 물질의 상업화를 위한 기초자료로 이용될 수 있을 것으로 사료된다.

울릉 마가목의 클로로겐산 이성체의 최종당화산물의 생성 저해 및 라디칼 소거 활성 (Chlorogenic Acid Isomers from Sorbus commixta of Ulleung Island Origin and Their Inhibitory Effects against Advanced Glycation End Product (AGE) Formation and Radical Scavenging Activity)

  • 김태훈
    • 한국식품영양과학회지
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    • 제45권8호
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    • pp.1208-1213
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    • 2016
  • 천연물로부터 당뇨합병증에 효과적인 소재를 개발하기 위하여 본 연구를 수행하였으며, 울릉도산 마가목 열매의 80% methyl alcohol 추출물의 ethyl acetate 가용부로부터 최종 당화산물 생성 억제능($IC_{50}$; $181.2{\pm}2.8{\mu}g/mL$)을 확인하였다. 효능성분의 동정을 위하여 $C_{18}$ 겔 등을 활용한 column chromatography를 수행하여 3종의 페놀성 화합물을 분리하였고, 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 neochlorogenic acid(1), cryptochlorogenic acid(2) 및 chlorogenic acid (3)로 동정하였다. 이들 화합물에 대해 최종당화산물 생성억제능을 평가한 결과 neochlorogenic acid(1)가 가장 강한 $167.5{\pm}3.5{\mu}M$$IC_{50}$ 값을 나타내었고, cryptochlorogenic acid(2)는 $185.9{\pm}3.7{\mu}M$$IC_{50}$ 값을 나타내었다. 또한, 이들 물질에 대해 $ONOO^-$ 소거 활성을 평가한 결과 cryptochlorogenic acid(2)가 가장 강한 라디칼 소거 활성인 $4.0{\pm}0.2{\mu}M$$IC_{50}$ 값을 나타내었고, 구조이성질체인 neochlorogenic acid(1)가 $4.5{\pm}0.3{\mu}M$$IC_{50}$ 값을 나타내었으며, 이들 활성은 caffeic acid가 결합 양상에 따른 화합물의 구조에 따라 다름이 시사되었다. 향후 이들 활성 물질의 활성 기작에 대한 연구가 필요하며 본 연구 결과는 더욱 우수한 최종당화산물 생성 억제능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을 뿐만 아니라 당뇨합병증 예방 및 치료에 효과적인 천연물질의 상업화를 위한 기초자료로 이용될 수 있을 것으로 생각한다.

Ampicillin의 경구투여에 따른 양식 어류(넙치, 조피볼락, 참돔)의 근육조직내 잔류량의 변화 (Muscle tissue Distribution Level of Ampicillin in Olive flounder(Paralichthys olivaceus), Rockfish(Sebastes schlegeli), and Red sea bream(Pagrus major) following oral administration)

  • 조윤희;정원철;신용운;김경원;하지영;허성혁;김의경;정희식;강석중;최유정;김석;이후장
    • 한국임상수의학회지
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    • 제23권2호
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    • pp.164-168
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    • 2006
  • The residue depletion of ampicillin was investigated in the olive flounder (Paralichthys olivaceus), rockfish (Sebastes schlegeli), and red sea bream (Pagrus major) after 5 days treatment with medicated feed at a dose of 100 mg/kg bw/day. Fishes were sampled for muscle on 1st, 2nd, 3rd, and 4th day after treatment. Ampicillin concentrations were determined by high performance liquid chromatography after SPE column extraction. The recovery rates of ampicillin in muscle samples ranged 94-98% and 83-88% for the concentration of 0.05 mg/kg and 0.1 mg/kg, respectively. Ampicillin concentrations detected on 1st day after treatment were 0.143, 0.138, and 0.187 mg/kg in the muscle of olive flounder, rockfish, and red sea bream, respectively. After a withdrawal of 3 days, muscle concentrations were 0.016, 0.012, and 0.021 mg/kg in the olive flounder, rockfish, and red sea bream, respectively. Ampicillin was not detectable in muscle samples on 4 days following withdrawal of the medicated feed. From results of the present study, a withdrawal period of ampicillin is proposed on 5 days after 5 days treatment with medicated feed at a dose of 100 mg/kg bw/day to avoid the presence of excessive residues of the edible muscles of olive flounder, rockfish, and red sea bream.

오매(烏梅)의 다성분 동시분석 및 항알러지 효과 (Simultaneous Analysis and Anti-allergic Effect of Mume Fructus)

  • 서창섭;하혜경;이호영;이준경;정다영;이진아;신현규
    • 생약학회지
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    • 제43권4호
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    • pp.279-285
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    • 2012
  • The Mume Fructus (MF) has been used for relieves cough, arrests arrest chronic diarrhea, treat fluid depletion, and treat ascariasis in Korea. In this study, a high-performance liquid chromatography (HPLC) method was established for simultaneous determination of six main components of MF. Additionally, we were investigated the anti-inflammatory and anti-allergic effects of MF extract on lipopolysaccharide (LPS)-treated RAW264.7 cells and tumor necrosis factor (TNF)-${\alpha}$/interferon (IFN)-${\gamma}$-treated HaCaT cells. The analytical column for separation was used a Gemini $C_{18}$ column maintained at $40^{\circ}C$. The mobile phase consisted of 1.0% (v/v) acetic acid in water (A) and 1.0% (v/v) acetic acid in acetonitrile (B). The flow rate was 1.0 mL/min and the detector was a photodiode array (PDA) set at 280 nm and 320 nm. We evaluated the inhibitory effect of MF extract on the production of inflammatory markers, nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in LPS-stimulated RAW264.7 cells and thymus- and activation-regulated chemokine (TARC/CCL17) in TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT cells, respectively. We confirmed the genes expression related with TARC, macrophage-derived chemokine (MDC/CCL22) and regulated on activation, normal T cell expressed and secreted (RANTES/CCL5) in HaCaT keratinocyte cells by MF extract. The contents of the five compounds in MF were 0.22-1.01 mg/g. Also, the MF extract show inhibition of about 78% and 75% on NO and $PGE_2$ production at the concentration 1000 mg/mL in RAW264.7 cells. MF extract suppressed the hTARC level and genes expression such as TARC, MDC, and RANTES on TNF-${\alpha}$/IFN-${\gamma}$-treated HaCaT cells.