• 제목/요약/키워드: HPLC screening

검색결과 159건 처리시간 0.031초

근적외선분광분석기를 이용한 미강의 Tocopherol과 Tocotrienol 함량 분석 (Quantification of Tocopherol and Tocotrienol Content in Rice Bran by Near Infrated Reflectance Spectroscopy)

  • 김용호;강창성;이영상
    • 한국작물학회지
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    • 제49권3호
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    • pp.211-215
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    • 2004
  • 미강에 함유되어 있는 토코페롤 및 토코트리에놀의 함량을 비파괴적으로 신속하게 추정하기 위하여 NIRS(근적외선 분광분석기)를 이용한 분석 방법을 검토하였다. 벼 유전자원 80계통의 미장을 사용하여 HPLC에서 분석된 토코페롤 및 토코트리에놀의 함량치를 NIRS 스펙트럼에 적용시킨 후 검량식을 작성하였다. NIRS의 검량식을 몇가지 방법에 의하여 비교 분석한 결과 2차 미분된 스펙트럼을 MPLS(Modified Partial Least Squares)를 이용한 회귀식에 이용하는 것이 가장 적합하였다. HPLC를 이용한 유전자원들의 성분 함량과 NIRS에서 도출된 검량식과의 상관계수는 토코페롤과 토코트리에놀이 각각 0.992, 0.953을 나타내었다. 이들 검량식은 validation file 에서도 0.846 및 0.956의 높은 상관을 보여 미강 상태에서 토코페롤 및 토코트리에놀의 함량을 NIRS를 이용하여 신속하게 분석할 수 있을 것으로 판단되었다.

High-Performance Liquid Chromatographic Quantification and Validation of Luteolin Glycosides from Sonchus brachyotus and Their Peroxynitrite-Scavenging Activity

  • Nugroho, Agung;Kim, Myung-Hoe;Lee, Chan-Mi;Choi, Jae-Sue;Lee, Sang-Hyun;Park, Hee-Juhn
    • Natural Product Sciences
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    • 제18권1호
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    • pp.39-46
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    • 2012
  • In Korea, the leaves of Sonchus brachyotus (Compositae), an edible mountainous vegetable, are traditionally used to treat hepatitis and hemorrhage and are known to have diuretic action. The aqueous ethanolic extract of this plant was selected in our screening experiment using the peroxynitrite ($ONO_2^-$)-scavenging assay, and the present study was performed to qualitatively and quantitatively identify the active compounds from S. brachyotus and validate the present high-permormance liquid chromatography (HPLC) coupled with ultraviolet absorption detection method based on accuracy, precision and repeatability. Five phenolic substances including the main compound, luteolin $7-O-{\beta}-D$-glucuronopyranoside, as well as chlorogenic acid, luteolin 7-O-rutinoside, luteolin $7-O-{\beta}-D$-glucopyranoside, and luteolin, were found in the aqueous ethanolic extract of S. brachyotus. In the HPLC validation experiment, the linearity of the four compounds was established by $R^2$ values of more than 0.999 within the test ranges, and the recovery rate ranged from 98.2 - 105.3%. Luteolin 7-O-glucuronide was a predominant compound (143 mg/g of extract and 18.3 mg/g of the dry weight of plant material) with a potent peroxynitrite-scavenging effect ($IC_{50}$, $1.02{\pm}0.08{\mu}M$). Luteolin and its three glycosides together with chlorogenic acid were qualitatively and quantitatively determined using an HPLC method validated in the present study.

Identification and Characterization of a Novel Antioxidant Peptide from Bovine Skim Milk Fermented by Lactococcus lactis SL6

  • Kim, Sang Hoon;Lee, Ji Yoon;Balolong, Marilen P.;Kim, Jin-Eung;Paik, Hyun-Dong;Kang, Dae-Kyung
    • 한국축산식품학회지
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    • 제37권3호
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    • pp.402-409
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    • 2017
  • A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

HPLC와 LC/MS에 의한 식육내 잔류 설파제의 동시 분석법 (Simultaneous analysis of sulfonamides in beef and pork by high performance liquid chromatography and electrospray ionization mass spectrometry)

  • 정봉수;박준조;금모래;김인경;박병옥;한정희
    • 한국동물위생학회지
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    • 제27권1호
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    • pp.17-29
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    • 2004
  • A multiresidual analysis was performed to determine 12 sulfonamides(sulfacetamide, sulfadiazine, sulfisomidine, sulfathiazole, sulfapyridine, sulfamerazine, sulfamethazine, sulfamonomethoxine, sulfisoxazole, sulfamethoxazole, sulfaquinoxaline, and sulfadimethoxine) in beef and pork simultaneously. The multiresidual analysis for the sulfonamides currently used was able to analyze 5 kinds of sulfonamides at the same time. The method of this 12 sulfonamides multiresidual analysis in this study was matrix solid-phase dispersion(MSPD) by high performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS). The recovery rate of the materials was measured by MSPD method with 3 different extraction solvents; Dichloromethane, DCM: Ethylacetate(3:1), DCM:EA(9:1). Also, samples (84 beef and 205 pork samples) which were positive by EEC-4 plate test from 2001 to 2003 were tested to investigate the kinds of sulfonamides using HPLC. The results from the study were as follows; 1. The recovery rate of the materials was measured by MSPD method with 3 different extraction solvents; Dichloromethane, DCM:Ethylacetate(3:1), DCM:EA(9:1). The method of extraction solvent with DCM:ethyl acetate(9:1) was the most excellent(87.7∼99.3%) in separation and reappearance. 2. In the LC/MS analysis. of sulfonamides, signal to noise ratio was showed relatively high in the positive mode and special ion in the quality analysis was determined via [M+H]$\^$+/ and m/z 156. A spectrum of sulfonamides was showed from all 12 sulfonamides. 3. The samples positive by the EEC-4 plate, a screening test method, were categorized by sulfonamides through Charm II and confirmed the kinds of sulfonamides through HPLC. 1) Among 84 beef samples positive by EEC-4 plate, 20 samples were positive by Charm II and identified as 7 sulfamethazine, 9 sulfadimethoxine, 1 sulfamonomethoxine and 3 unknown status. 2) Among 205 pork samples positive by EEC-4 plate, 42 samples were positive by Charm II and identified as 19 sulfamethazine, 1 sulfadimethoxine, 4 sulfamonomethoxine and 5 unknown status.

Development of Immunochromatography Strip-Test Using Nanocolloidal Gold-Antibody Probe for the Rapid Detection of Aflatoxin B1 in Grain and Feed Samples

  • Shim, Won-Bo;Yang, Zheng-You;Kim, Jung-Sook;Kim, Ji-Young;Kang, Sung-Jo;Woo, Gun-Jo;Chung, Young-Chul;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • 제17권10호
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    • pp.1629-1637
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    • 2007
  • An immunochromatography (ICG) strip test using a nanocolloidal gold-antibody probe was developed and optimized for the rapid detection of aflatoxin B1 (AFB1). A monoclonal antibody specific to AFB1 was produced from the cloned hybridoma cell (AF78), coupled with nanocolloidal gold, and distributed on the conjugate pad of the ICG strip test. The visual detection limit of the ICG strip test was 0.5 ng/ml, and this method showed a cross-reaction to aflatoxin B2, G1, and G2. In total, 172 grain and feed samples were collected and analyzed by both the ICG strip test and HPLC. The results of the ICG strip test showed a good agreement with those obtained by HPLC. These results indicated that the ICG strip test has a potential use as a rapid and cost-effective screening tool for the determination of AFB1 in real samples and could be applied to the preliminary screening of mycotoxin in food and agricultural products, generating results within 15 min without complicated steps.

호밀, 귀리 추출물로부터 제초활성물질 탐색 및 동정 (Screening and Identification of Natural Herbicidal Active Substance in Rye and Oat Extracts)

  • 양계진;김광호;정일민
    • 한국잡초학회지
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    • 제18권4호
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    • pp.348-355
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    • 1998
  • 고소득 작물인 인삼의 생산비 절감 및 고품질화를 위한 새로운 재배법을 확립하기 위하여 Allelopathic 식물이 함유하고 있는 제초활성물질 탐색 동정한 결과는 다음과 같다. 12종류의 표준품을 이용하여 호밀과 귀리의 추출물에 대한 Allelopathy 물질을 분석한 결과, 두 추출물간에 함유하는 물질의 차이를 보였으나 호밀에서는 benzoic acids계열인 salicylic acid(8.34mg/g), 귀리에서는 flavonone glycosides계열인 naringin(7.50mg/g)이 가장 많이 함유되어 있음을 알 수 있었으며, 동정된 표준품을 이용하여 $10^{-3}M$$10^{-4}M$에서 명아주 종자에 대한 발아율, 평균 발아 일수 및 발아세를 조사한 결과 대조구에 비하여 통계적으로 낮은 값을 보여 salicylic acid, naringin를 비롯하여 이러한 물질들이 Allelopathy를 나타내는 성분일 것으로 생각되었다.

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감자 더뎅이병원균에 대해 길항활성을 갖는 방선균 탐색 및 항균 활성물질의 분리 (Screening of Antagonistic Actinomycetes for Potato Scab Control and Isolation of Antibiotic Compound)

  • 이향범;조종운;임치환;김창진
    • Applied Biological Chemistry
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    • 제47권2호
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    • pp.164-169
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    • 2004
  • 감자더뎅이병 방제용 생물제제(biocontrol agenL BCA)를 개발하기 위하여 국내 토양으로부터 분리된 5,000여 방선균 균주를 대상으로 더뎅이병 관련 병원균(Streptomyces scabiei및 S. turgidiscabies)에 대한 in vitro또는 in vivo 활성검정을 실시하였다. 활성검정 결과 길항력이 우수한 균주로서 9균주가 선발되었으며 실제 재배포장에서 사용되고 있는 dazomet및 mancozeb 등 농약에 대한 감수성 시험을 실시하여 A020645 균주가 길항활성 뿐만 아니라 약제저항성이 가장 높음을 확인하고 더뎅이병 방제용 BCA 균주로 선발하였다. 본 균주로부터 항균활성물질을 분리하기 위하여 액체배양액으로부터 음이온교환 크로마토그래피(anion exchange chromatography), solidphase(ODS) extraction, TLC, 역상 HPLC 등을 실시하여 최종적으로 compound A와 B를 순수 분리하였다. Compound A와 B는 NMR 분석 결과 nucleoside계 화합물로 판단된다.

Aflacxin에 대한 최신 분석법과 규제동향 (Development of Rapid, Safe Analytical Techniques of Aflatoxins and Their Current Regulation)

  • 정덕화
    • 한국식품위생안전성학회지
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    • 제5권3호
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    • pp.131-138
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    • 1990
  • Aflatoxins is a chemically diverse group of toxic secondary metabolites that are produced by fungi and often occur in agricultural commodities. Because of their wide range of toxic effects, Aflatoxins cause severe economic losses to farmers and livestock producers and pose a health to human consuming contaminated foods. Long term prospects for biotechnological control of Aflatoxins require elucidation of the specific steps and regulation of their biosynthetic pathways . Aflatoxin determinations can be approached many ways. It is essential to safely handle all experimental materials associated with aflatoxin analysis or aflatoxigenic fungi Visual screening of suspect samples, base on the presence of conidial head of the aspergillus flavus group, and screening samples for the presence of bright greenish yellow flourescence are not chemical tests and such screening techniques may allow aflactoxin contaminated lots into commerce. Microcolumn screening procedures should always be used in conjunction with a quantitative method. Several thin layer chromatography(TLC) and high performance liquid chromatography(HPLC) methods are suitable for quantitation and are in general use. Immunochemical Methods such as the ELISA or affinity column chromatography methods are being rapidly developed. The chemical and immunochemical methods can be reliable if care is taken, using suitable controls and personnel that are well trained . All analytical laboratories should stress safety and include suitable analytical validation procedure. Especially a worldwide enquiry was undertaken in recent to obtain up-to-date information about aflatoxin legislation in as many countries of the world as possible. The information concerns aflatoxin in foodstuffs. aflatoxin MI in dairy products, aflatoxins in animal feedstuffs. Limits and regulations for aflatoxin have been expended in recent with more countries having legislation on subject, more products, and more aflatoxins covered by this legislation.

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Ligustici Rhizoma(고본(藁本))와 Angelica tenuissimae Radix(한국고본(韓國藁本)뿌리)의 규격화(規格化) 및 유효성분(有效成分)의 진통효과(鎭痛效果) 비교(比較)에 관한 연구(硏究) (A Study on the Standardization of Ligustici Rhizoma and Angelicae Tenuissimae Radix including the Comparison for the Biological Activity on the Active Ingredients)

  • 김관호;이상인;김호철;박호군;이재성
    • 대한한의학회지
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    • 제18권1호
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    • pp.187-197
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    • 1997
  • Major separation for the active ingredients and characterization of chemical properties in conjunction with screening test on animal were performed in order to analyze and standardize Ligustici Rhizoma or Angelicae Tenuissimae Radix as an important oriental herbal medicine for antiphlogistic or an important oriental herbal medicine for antiphlogistic or an anodyne. Furthermore the structure, composition and contents of ingredients for essential oil in Angelicae Tenuissimae Radix(Suckpo, Korea) were determined by means of Ge/MS followed by screening test on Z-ligustilide(82%) known as major ingredient as well as butylidenephthalide collected by HPLC with normal phase semiprep-column. The total active ingredient in Ligustici Rhizoma from China or Angelicae Tenuissimae harvested at Choonyang(Kyungnam, Korea), Jungsun(Kangwon, Korea), Suckpo(Kyungnam Korea), Youngchun(Kyungnam, Korea) have been determined showing higher abundant for three times on the product in Korea compared to that in China. In addition, the major component in Ahgelicae Tebyussunae Radux extract was found to be Z-ligustilide(70-80%) which is very different from that in Ligustici Rhizoma senkyunolide(39%) as major species. For screening test of Ligustici Rhizoma or Angelicae Tenuissimae Radix extracts toward the target animal, the efficiency has been shown the similarity on both extracts. Taking into account the level of ingredient, the total efficiency may be three times higher on Angelicae Tenuissimae Radix in Korea compared to Ligustici Rhizoma in China. As a result of present study, it is preferable to distinguish between Ligustici Rhizoma and Angelicae Tenuissimae Radix for better usage of oriental herbal medicine because of very different composition and abundant in spite of their similar screening effect.

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Loop와 HPLC Purification 방법보다 더 높은 비방사능을 보여주는 카트리지 Methylation과 Purification을 이용한 손쉬운 [ 11C]PIB 합성 (Facile [11C]PIB Synthesis Using an On-cartridge Methylation and Purification Showed Higher Specific Activity than Conventional Method Using Loop and High Performance Liquid Chromatography Purification)

  • 이용석;조용현;이홍재;이윤상;정재민
    • 핵의학기술
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    • 제22권2호
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    • pp.67-73
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    • 2018
  • $[^{11}C]PIB$는 베타아밀로이드($A{\beta}\;plague$)라는 변성 단백질에 결합하여 뇌의 기능과 기억력을 서서히 감퇴시키는 비가역적인 질환인 치매를 조기에 감별할 수 있는 대표적인 방사성의약품이다. 지금까지 많은 실험실에서 $[^{11}C]PIB$는 자동화합성장치에서 $[^{11}C]methyl\;iodide$$[^{11}C]methyl\;triflate$를 만든 다음 loop나 vial 방법을 사용하여 methylation을 한 다음 HPLC로 정제를 하는 것이다. 하지만 기존의 보고된 방법은 시간이 오래 걸리며, HPLC와 같은 복잡한 시스템을 필요로 하여 소규모 실험실에서 합성하기에 적합하지 않으며, 최종 product에서 에탄올 함량이 높다는 단점이 있었다. 이러한 단점을 보완하기 위하여 카트리지만을 사용하여 카트리지에서 methylation과 purification을 동시에 실시함으로써 합성 시간을 단축하고, 비방사능이 높고, 낮은 에탄올 함량을 가진 $[^{11}C]PIB$를 합성 가능한지 확인하고자 하였다. 가장 널리 사용하는 카트리지 6종(CM, HLB, Alumina, C18, tC18, tC18 environmental을 선택하여 screening test를 실시하였다. 6-OH-BTA-0 1 mg을 c-HXO에 녹인 다음 6개의 카트리지에 loading를 한 다음 0.5 M MSP(pH 5.1) 20 mL로 정제를 한 다음 최종 fraction을 받아서 analytical HPLC로 전구체 잔류량을 측정한 결과 hydrophobicity가 낮은 계열(CM, HLB, Alumina)의 카트리지에서는 완충액으로 정제를 하였을 때 잔류전구체의 양이 많았으나, 탄소함량이 많은 계열의 카트리지(C18, tC18, tC18 environmental)에서는 잔류전구체의 양이 CM, HLB, Alumina 카트리지에 비하여 상대적으로 적었다. 완충액의 정제 농도와 부피를 최적화 하기 위하여 screening test에서 가장 좋은 결과를 나타낸 C18 series cartridge를 가지고 추가 실험을 진행하였다. 인산완충액 농도를 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 250 mM, 500 mM로 변화시켰으며, 에탄올 함량은 20%와 30%로 하여 용출액을 분석하여서, $[^{11}C]PIB$를 카트리지로 합성하기 위한 최적의 조합은 tC18 environmental cartridge와 0.5 M MSP 20 mL인 것을 알 수 있었다. 기존에 보고된 방법과 cartridge를 비교한 결과, 합성시간에서는 각각 15 ~ 18min, 8 ~ 9 min이 소요되었으며, product activity는 각각 $4.1{\pm}1.4\;GBq$ (n=41), $3.8{\pm}0.9\;GBq$ (n=3), 방사화학적 수율(based on HPLC analysis of the crude product)에서는 $13.9{\pm}4.4%$ (n=41), $12.3{\pm}2.2%$ (n=3)로 별다른 차이가 없었으며, 비방사능에 있어서는 HPLC purification method가 $78.7{\pm}39.7\;GBq/{\mu}mol$ (n=41), cartridge method가 $420.6{\pm}20.4\;GBq/{\mu}mol$ (n=3)로 카트리지 방법이 기존 방법보다 더 좋은 결과를 나타내었다. 또한, 잔류 용매(c-HXO)도 vial or loop method와 별다른 차이가 없었으며, 에탄올 함량에 있어서는 70%(기존 방법)에서 30%(카트리지 방법)로 두 배 이상 함량이 적다는 사실을 알 수 있었다. 지금까지 알아본바와 같이 cartridge method는 reported method(HPLC purification)에 비하여 더 향상된 결과를 보여준다는 사실을 확인하였다.