• 제목/요약/키워드: Reagent-based method

검색결과 73건 처리시간 0.025초

Selection of Tropane Alkaloids High-Producing Lines by Single Cell Cloning of Hyoscyamus niger L. Root Cultures

  • Min, Ji-Yun;Park, Dong-Jin;Jeong, Mi-Jin;Song, Hyun-Jin;Kang, Seung-Mi;Kang, Young-Min;Choi, Myung-Suk
    • 한국산림과학회지
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    • 제98권2호
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    • pp.142-147
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    • 2009
  • Hyoscyamus species is sources of the hypnotic and sedative drugs hyoscyamine and scopolamine. Single cells of Hyoscyamus niger were dissociated from suspension cultures and adventitious roots obtained from single-cell clones which were cultured on B5 medium containing 3% (w/v) sucrose, 0.1 mg/L IBA and 0.4% (w/v) gelrite. H. niger adventitious root lines showed wide variation in tropane alkaloids production and growth. An effective selection of 200 root lines was made possible by the application of the 'Dragendorff's reagent' for qualitative detection of the alkaloids from root. A high correlation coefficient (r=0.9390) was observed between the values obtained with the two methods based on HPLC and Dragendorff's reagent analysis. Among the selected roots, the highest scopolamine content was 16.64 mg/g DW (Hn-59), which was 8.82-fold more productive than the lowest alkaloid producing line (Hn-25). Here, we established an efficient selection method on tropane alkaloids production and suggest that the Dragendorff's reagent is of great practical value in selection of invisible compounds.

Aconitine 류 알칼로이드의 검출에 의한 부자류 생약의 확인 (Identification of Buza by Detecting Aconitine-type Alkaloids)

  • 엄동옥;반태환
    • 분석과학
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    • 제12권3호
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    • pp.243-247
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    • 1999
  • HCl 산성에서 aconitine류 알칼로이드는 Reineke 시액으로 백색의 침전을, 그리고 TCR 시액으로 청색의 침전을 형성하였다. 백색의 침전은 $NH_4OH$ 알칼리성의 물에 녹아 aconitine류 알칼로이드를 유리시키며, 유리된 aconitine류 알칼로이드는 DCE층으로 쉽게 전용되었다. 전용된 DCE층을 분취하여 UV흡광파장을 측정하고 TLC로 분리하였다. 청색의 침전은 DCE층에 전용되므로 전용된 DCE층을 분취하여 UV/Vis.흡광파장을 측정하였다. 이상의 방법으로 aconitine류 알칼로이드를 검출하여 생약이나 제제에서 aconitine류 알칼로이드를 함유한 부자류 생약을 확인할 수 있었다.

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Determination of Theophylline and its Metabolites in Human Urine by High-Performance Liquid Chromatography

  • Kim, Kyeong-Ho;Park, Young-Hwan;Park, Hyo-Kyung;Kim, Ho-Soon;Lee, Min-Hwa
    • Archives of Pharmacal Research
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    • 제19권5호
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    • pp.396-399
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    • 1996
  • High-performance liquid chromatographic method with UV detecction was developed for the determination of theophylline and its metabolites in human urine using ${beta}$-hydroxyethyl theophylline$({beta} -HET)$ as an internal standard. For extraction of urine sample, quality control sample and xanthine-free blank urine were mixed with decylamine (ion-paring reagent) and ${beta}$-HET. After saturation with ammonium sulfate, the mixture was then extracted with organic solvent at pH values of 4.0-4.5. All separations were performed with ion-pair chromatography using decylamine as an ion-pairing reagent and 3mM sodium acetate buffered mobile phase (pH 4.0) containing 1% (v/v) acetonitrile and 0.75 mM decylamine. The detection limits of theophylline, 1, 3-DMU, 1-MU, 3-MX and 1-MX in human urine were 0.17, 0.17, 0.39, 0.19 and 0.19 ${\mu}g$/ml, based on a signal-to-noise ratios of 3.0. The mean intraday coefficients of variation (C.V.s) of each compound on nine replicates were lower than 2.0%, while mean interday C.V.s on three days were lower than 1.6%. All separations were finished within 40miutes.

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음향적 흐름을 이용한 혈액형 분석을 위한 새로운 방법 (A novel Method for Blood Typing using Acoustic Streaming)

  • 최현주;장웅식;남정훈;임채승
    • 대한의용생체공학회:의공학회지
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    • 제39권6호
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    • pp.250-255
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    • 2018
  • Accurate blood typing is the crucial factor for safe and successful blood transfusion and plays a very important role in organ transplantation and genetic information of forensic medicine. Microfluidic devices have been developed to overcome the limitations of the conventional blood typing methods. In this study, we demonstrate a Lamb wave-based device for simple blood typing in a sample droplet and we propose new indices for quantitative and accurate blood typing. Using Lamb wave-induced acoustic streaming in the droplet, the blood sample and the reagent can be mixed rapidly and red blood cells start to form clumps, which is agglutination. Based on the recorded image and video, the intensity of transmitted light through the sample droplet is evaluated to determine the blood type. Effect of the concentration of suspended red blood cells was evaluated and we found that 10% concentration of suspended red blood cells was suitable to observe the difference between aggregated and non-aggregated samples. Finally, sample with blood type A could be determined using anti-A reagent in our Lamb wave-based device. Our device enables simple and accurate blood typing, which can be applied to resource-limited environments.

Silicon Carbide Coating on Graphite and Isotropic C/C Composite by Chemical Vapour Reaction

  • Manocha, L.M.;Patel, Bharat;Manocha, S.
    • Carbon letters
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    • 제8권2호
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    • pp.91-94
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    • 2007
  • The application of Carbon and graphite based materials in unprotected environment is limited to a temperature of $450^{\circ}C$ or so because of their susceptibility to oxidation at this temperature and higher. To over come these obstacles a low cost chemical vapour reaction process (CVR) was developed to give crystalline and high purity SiC coating on graphite and isotropic C/C composite. CVR is most effective carbothermal reduction method for conversation of a few micron of carbon layer to SiC. In the CVR method, a sic conversation layer is formed by reaction between carbon and gaseous reagent silicon monoxide at high temperature. Characterization of SiC coating was carried out using SEM. The other properties studied were hardness density and conversion efficiency.

Quantitative Analysis of the Degree of Silanization by the Ninhydrin Method and its Application to the Immobilization of GL-7-ACA Acylase and Cellulolytic Enzyme

  • Park, Seung-Won;Kim, Yong-In;Chung, Koo-Hun;Kim, Seung-Wook
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.199-203
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    • 2001
  • A simple quantitative method to measure the degree of silanization was developed, based on the reaction of ninhydrin with the silanization reagent (3-aminopropyltriethoxysilane, 3-APTES). At low concentrations (0.001-0.005%, v/v) of 3-APTES, a good linearity was obtained when 3-APTES reacted with undiluted ninhydrin for 30 min. On the other hand, at high levels of 3-APTES, a linearity was obtained when 3-APTES reacted with 3-fold diluted ninhydrin for 20 min. The reliability of regression curves mentioned above was expressed as a regression coefficient ($R^2$) of more than 0.99. Immobilization of different enzymes was introduced via silanization by using the 3-APTES in order to confirm the validity of the ninhydrin method. When yield for each step in the immobilizatio process were compared, yields of both glutaraldehyde and protein were founc to have the same tendency to silanization. These results shw that the ninhydrin method was suitable for quatitative analysis of silanization and that yields of immobilization could be pre-estimated by measuring silanization levels using the ninhydrin method.

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Application of Organized Media for Rapid Spectrofluorimetric Determination of Trace Amounts of Cr(VI) in the Presence of Cr(III)

  • Madrakian, Tayyebeh;Afkhami, Abbas;Mohammadnejad, Masoumeh
    • Bulletin of the Korean Chemical Society
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    • 제30권6호
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    • pp.1252-1256
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    • 2009
  • A simple, selective and sensitive fluorescence quenching method was developed to the determination Cr(VI). The method is based on the oxidation of $I^-\;to\;{{I_3}^-}$ by Cr(VI) in sulfuric acid solution followed by immediate formation of ion association compound between I3 − and rhodamine 6G in Tween-80 micellar media at room temperature. The influence of several surfactants on rhodamine 6G fluorescence signal was studied; particular attention was paid in the aggregation behavior of rhodamine 6G–Tween-80 system. The experimental parameters (e.g., type of surfactant, reagent concentration) were studied and the optimal conditions were established. The linear calibration graph was obtained in the range 2.0 - 100.0 ng m$L^{-1}$ Cr(VI). The detection limit of the method was 0.37 ng m$L^{-1}$. The relative standard deviation (R.S.D.) is less than 5% (n = 5). The efficiency of the method for the determination of Cr(VI) in the presence of Cr(III) in the sample was investigated. The method was applied successfully to the determination of Cr(VI) and total Cr in water, and liver tissue samples.

Cyanide detection based on natural dyes reaction from blue butterfly pea flowers (Clitoria Ternatea)

  • Chotichayapong, Chatrachatchaya;Kuchaiyaphum, Pusita;Butwong, Nutthaya;Bua-ngern, Worapong
    • 분석과학
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    • 제35권4호
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    • pp.153-160
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    • 2022
  • A green spectrophotometric method for the determination of cyanide has been proposed using, a green reagent, aqueous extract of blue butterfly pea. The test tube was filled with anthocyanin rich extract (pH 6) and cyanide solution. The reaction was kept constant for 10 minutes at room temperature. The reaction mixture changed color from blue to green as the amount of CN-ions increased. The 620 nm peak intensity increased with CN concentration. Therefore, this wavelength was used for all cyanide analyses. The cyanide calibration curve had a linear range of 0.25-1.00, 1.00-4.00, and 4.00-10.00 mg/L, with a satisfactory correlation coefficient of 0.99 and a LOD of 0.57 mg/L. The recovery ranged from 8.33 to 76.94 percent, indicating that this method is inaccurate at low cyanide concentrations. The intra-day and intermediate precision relative deviations were 0.391-0.871 % and 1.112-1.583 %. An H-bond forms between the C-4 group of the B-carbonyl ring and the HCN molecule according to the B3LYP/TZVP calculation. The method is convenient for cyanide concentrations above the LOQ of 1.09 mg/L, cost-effective, and capable of reducing toxic solvents with acceptable precision. The method could also be used to detect total cyanide in biological, environmental, and industrial waste samples.

검사별 radioimmunoassay시약 조사 및 비교실험 (Radioimmunoassay Reagent Survey and Evaluation)

  • 김지나;안재석;전영우;윤상혁;김윤철
    • 핵의학기술
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    • 제25권1호
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    • pp.34-40
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    • 2021
  • [목 적] 의료기관의 핵의학 검사실에서 신규검사를 도입하거나 사용하던 시약을 변경하게 되는 경우 절차에 따라 검사의 특성이 분석되고 시약에 대한 평가가 이루어져야 한다. 그러나 요구되어지는 비교실험을 모두 수행하기 위해서는 몇 가지 필요한 조건이 충족되어야 하는데, 첫째 각 검사별로 수행하기에 충분한 검체량이 준비되어야하며, 둘째 비교실험에 적용 가능한 다양한 시약의 공급이 가능해야한다. 충분한 비교실험이 이루어졌다고 하더라도 변경된 시약에 의한 데이터 변동이 전체 환자데이터 변동을 의미하는 것에는 한계가 있으므로 검사실에서 시약이 변경되는 것에 대한 부담이 있다. 이러한 다양한 어려움으로 검사실에서의 시약변경은 제한적으로 이루어지고 있다. 본원에서는 원할한 경쟁 입찰을 도입하기 위하여 검사별로 radioimmunoassay(RIA)시약을 전수조사하고 비교실험을 통해 검사실에서 사용가능한 시약범위를 설정하였다. 이 과정을 공유하고자 하였다. [대상 및 방법] 본원 핵의학 검체 검사실에서 시행하고 있는 검사는 위탁검사를 제외하고 총 20종목이다. 각각의 검사별로 외부정도관리와 기관간 정도관리 결과보고서를 참고로 사용가능한 RIA시약을 전수 조사하였고, 각 시약에 대한 메뉴얼을 확보하였다. 각각의 시약마다 메뉴얼을 확인하여 검사 방법과 incubation시간, 검사 시 필요한 검체량, 시약량 등을 확인하여 본 검사실에서 사용가능한지 여부에 따라 시약 1차 선정을 하였다. 1차 선정된 시약을 100 test기준으로 2 kit씩 공급받아 데이터 상관성시험, 민감도, 회수율, 희석시험을 진행하였고, 비교실험 결과에 따라 시약을 2차 선정하였다. 1, 2차 선정을 통과한 시약을 경쟁 입찰리스트로 제출하였다. 검사 시약을 단수로 지정할 경우에는 1차, 2차 선정 과정에서 얻은 자료로 단수지정 사유서를 작성하였다. [결 과] 각각의 시약마다 매뉴얼을 확인하여 시약 1차 선정에서 제외되는 경우는 각 검사의 현재 Turn Around Time(TAT)보다 길어지는 경우와 검사 시 사용 시약량이 많아 장비사용이 불가능한 경우였다. 1차 선정에서 사용가능한 시약이 1개인 경우는 5종목 squamous cell carcinoma antigen(SCC Ag), 𝛽-human chorionic gonadotropin(𝛽-HCG), vitamin B12, folate, free testosterone 이었고, 2개인 경우는 8종목 (CA19-9, CA125, CA72-4, ferritin, thyroglobulin antibody(TG Ab), microsomal antibody(Mic Ab), thyroid stimulating hormone-receptor-antibody(TSH-R-Ab), calcitonin), 3개인 경우는 5종목(triiodothyronine(T3), Free T3, Free T4, TSH, intact parathyroid hormone(intact PTH)), 4개인 경우는 2종목(carcinoembryonic antigen(CEA), TG)이었다. 2차 최종 선정결과 사용가능한 시약이 3개인 것은 T3, Free T3, Free T4, TSH, CEA, 2개인 것은 TG Ab, Mic Ab, TSH-R-Ab, CA125, CA72-4, intact PTH, calcitonin이었다. 단수 지정된 종목은 ferritin, TG, CA19-9, SCC, 𝛽-HCG, vitamin B12, folate, free testosterone이었다. 2차 선정에서 제외된 사유에는 비교실험을 위한 시약공급이 안된 경우와 데이터 재현성에 문제가 있었던 경우, 데이터 변동에 대한 수용이 불가능하다고 판단되는 경우였다. 비교실험 시 가장 문제가 되는 부분은 검체 수집이었다. 검사건수가 많고 검사 시 필요한 검체량이 적은 경우에는 문제가 되지 않았지만, 검사건수가 적은 경우(월 100건 이하)에는 다양한 농도 검체를 수집하기가 어려웠으며, 한번 검사 시 필요한 검체량이 상대적으로 많은 경우(100 uL이상)에는 회수율시험을 진행하기가 어려웠다. 또한 민감도 측정이나 희석시험을 위한 희석액이나 표준액0 물질이 부족한 경우도 문제점 중의 하나였다. [결 론] 검사시약 변경을 위한 비교실험 시 다양하고 충분한 검체 수집을 위해 적정한 준비기간이 필요하다. 또한 1회 검사 시 필요한 검체량 및 시약량에 따라 비교실험 시 필요한 총 검체량, 시약량 범위를 설정해 놓는다면 비교실험을 진행할 때마다 검체 수집과 실험계획을 세우는 데 부담이 줄어들 것이다.

Colloidal Textile Dye-Based Dipstick Immunoassay for the Detection of Infectious Flacherie of Silkworm, Bombyx mori L.

  • Sivaprasad, V.;Nataraju, B.;Renu, S.;Datta, R.K.
    • International Journal of Industrial Entomology and Biomaterials
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    • 제6권1호
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    • pp.27-31
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    • 2003
  • Infectious flacherie of silkworm Bombyx mori is caused by B. mori infectious flacherie virus (BmIFV) and causes severe crop loss to sericulturists. In the present study, a colloidal textile dye-based dipstick immunoassay is developed for the detection of infectious flacherie in silkworms. Colloidal textile dye (blue D2R) with Aλ$_{max}$ at 620 nm was sensitised with 500 $\mu\textrm{g}$/ml of purified anti-BmIFV IgG. The dye-antibody reagent detects purified antigen up to 10 ng/ml and BmIFV infection in diseased larval extracts $(up to a dilution of {10^-5})$ and faecal matter extracts $(up to a dilution of {10^-2})$ by forming clear blue dot within 30 min. It was observed to be stable for three months period at $4^{\circ}C$. The efficacy of textile dye-based dipstick immunoassay was on pay with HRP-based dipstick immunoassay and fluorescent antibody test, and better than latex agglutination and ouchterlony tests in the detection of BmIFV The dye-based dipstick immunoassay method provides a simple, sensitive and less expensive test for the detection of BmIFV infection in silkworms.s.