• Title/Summary/Keyword: Colorimetric Assay

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Estimating dehalogenation reactivity of nanoscale zero-valent iron by simple colorimetric assay by way of 4-chlorophenol reduction

  • Mines, Paul D.;Kaarsholm, Kamilla M.S.;Droumpali, Ariadni;Andersen, Henrik R.;Hwang, Yuhoon
    • Environmental Engineering Research
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    • v.25 no.2
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    • pp.197-204
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    • 2020
  • A number of different nanoscale zero-valent iron (nZVI) materials have been prepared and compared depending on the desired properties for the particular application, but different physicochemical properties of this prepared nZVI make it difficult to universally compare and standardize them to the same scale. In this study, we aimed to demonstrate a simple microplate-based colorimetric assay using 4-chlorophenol as an indicator with respect to the remediation of real treatment targets, such as trichloroethylene (TCE), 1,1,1-trichloroethane (TCA), and atrazine. Effect of nickel contents on 4-chlorophenol reduction was successfully investigated by the miniaturized colorimetric assay. In the same manner, the effect of nickel contents on dehalogenation of TCE, TCA, and atrazine was investigated and the pseudo-first-order kinetic constants were compared with the results for 4-chlorophenol. The similar pattern could be observed between 4-chlorophenol reduction obtained by colorimetric assay and TCE, TCA, atrazine reduction obtained by a traditional chromatographic method. The reaction kinetics does not match perfectly, but the degree of reaction can be estimated. Therefore, the colorimetric assay can be a useful and simple screening tool to determine nZVI reactivity toward halogenated organics before it is applied to a particular remediation site.

Cytotoxicity Evaluation of Cosmetic Materials to Mouse Fibroblast : by Tetrazolium salt, MTT Colorimetric Assay (Tetrazolium salt, MTT Colorimetric Assay를 이용한 Mouse Fibroblast에 대한 화장품원료 물질의 세포독성 평가)

  • Jo, Jae- Hoon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.15 no.1
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    • pp.37-50
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    • 1989
  • The in Vitro chemosensitivity of fibroblast cell strains was determined using a semiautomated tetrazolium-based colorimetric assay(MTT assay) to 16 cosmetic materials. This assay is useful method to evaluate toxic effects of the chemicals. From assay results, we determined that the preservatives are more toxic than moisteurizers. The chemicals in the same group have a different toxicity. That is, in preservatives, Germall -115 is more toxic than Danisol -M, -p, and in surfactant sodium laurel sulfate than Myrj 52, and in moisteurizers, 1, 3-butylene glycol is more safe than the others. When the results from this assay for preservatives were compared with patch test results, good correlation was observed. Forthemore, this assay method can be used together with Patch test for the evaluation of the chemical toxicity, particularly in cosmetic field.

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Direct Colorimetric Assay of Microcystin Using Protein Phosphatase

  • Oh, Hee-Mock;Lee, Seog-June;Kim, Jee-Hwan;Park, Chan-Sun;Yoon, Byung-Dae
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.418-421
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    • 2000
  • A new direct colorimetric assay of microcystin in water and algal samples is proposed consisting of two procedures as follows: 1) the elimination of phosphorus in the sample and concentration of microcystin using a C(sub)18 cartridge, 2) the detection of the released phosphorus by the ascorbic acid method and determination of protein phosphatase (PP) inhibition by microcystin. The optimum amounts of phosphorylase ${\alpha}$ and PP-1 in 50 ${\mu}$L concentrated sample were 50$\mu\textrm{g}$/50${\mu}$L buffer and 1.0unit/50${\mu}$L buffer, respectively, for the best assay. The pH for the maximum activity of PP-1 was 8. The minimum detectable concentration for this method was about 0.02$\mu\textrm{g}$/L, which is sufficient to meet the proposed guideline level of 1$\mu\textrm{g}$ microcystin/L in drinking water. Consequently, it would seem that the proposed direct colorimetric assay using PP is a rapid, easy, and convenient method for the detection of microcystin in water and algal samples.

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Evaluation of Microbial Epoxide Hydrolase Activity Based on Colorimetric Assay Using 4-(p-nitrobenzyl) Pyridine (4-(p-Nitrobenzyl)pyridine의 색깔반응을 이용한 미생물 epoxide hydrolase의 활성 평가)

  • Kim Hee Sook;Lee Eun Yeol
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.332-336
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    • 2005
  • Epoxide hydrolase activities of various microbial cells were analyzed by colorimetric assay based on alkylation of epoxides with 4-(p-nitrobenzyl)pyridine (NBP). The epoxide hydrolase activity was determined by measuring the decrease of color intensity at 560 nm due to the decrease of styrene oxide substrate by epoxide hydrolase-catalyzed hydrolysis reaction. The experimental conditions of NBP colorimetric assay were optimized for the efficient measurement of epoxide hydrolase activities from various microbial cells.

Development of a Redox Dye-Based Rapid Colorimetric Assay for the Quantitation of Viability/Mortality of Pine Wilt Nematode

  • Han, Kyeongmin;Lee, Jaejoon;Shanmugam, Gnanendra;Lee, Sun Keun;Jeon, Junhyun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1117-1123
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    • 2019
  • Control of pine wilt disease, which is caused by pine wilt nematode Bursaphelenchus xylophilus, is heavily dependent on the use of chemicals such as abamectin. Although such chemicals are highly effective, demands for alternatives that are derived preferentially from natural sources, are increasing out of environmental concerns. One of the challenges to discovery of alternative control agents is lack of fast and efficient screening method that can be used in a high-throughput manner. Here we described the development of colorimetric assay for the rapid and accurate screening of candidate nematicidal compounds/biologics targeting B. xylophilus. Contrary to the conventional method, which relies on laborious visual inspection and counting of nematode population under microscope, our method utilizes a redox dye that changes its color in response to metabolic activity of nematode population in a given sample. In this work, we optimized parameters of our colorimetric assay including number of nematodes and amount of redox dye, and tested applicability of our assay for screening of chemicals and biologics. We demonstrated that our colorimetric assay can be applied to rapid and accurate quantification of nematode viability/mortality in a nematode population treated with candidate chemicals/biologics. Application of our method would facilitate high-throughput endeavors aiming at finding environment-friendly control agents for deadly disease of pine trees.

A Colorimetric Glucose Assay via Concentration Gradient Paper Chip (종이기반 농도 구배 형성 칩을 통한 포도당 발색 반응 검사)

  • Kim, Taehoon H.;Shin, Hyun Young;Lee, Yun-Il;Tae, Ki-Sik;Kim, Minseok S.
    • Journal of Biomedical Engineering Research
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    • v.38 no.6
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    • pp.302-307
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    • 2017
  • This paper presents a paper-based concentration gradient chip to analyze colorimetric glucose assay. The paper-based concentration gradient chip was fabricated through a wax patterning technique that can design the fluidic channel by selectively printing hydrophobic and hydrophilic areas. Afterwards, glucose and dilution solutions were loaded into the inlet of a concentration gradient chip and each solution was then mixed sequentially at mixing channel. Finally, concentration gradient was formed at each outlet of the chip. To measure the glucose concentration of the solution in outlets, we conducted colorimetric glucose assay with fixed concentration of glucose solution (0, 5, 10, 15 and 20 mM) and obtained normalized intensity. Subsequently, glucose concentrations of the outlets were calculated by substituting the normalized intensity to linear regression function based on the normalized intensity of fixed glucose concentration. Finally, the concentration gradient of glucose was formed on the chip with the result of colorimetric assay. The concentration gradient paper chip has the potential to accurately analyze unknown glucose concentration.

Cytotoxic Effect of Syringic Acid on Human Oral Epithelioid Carcinoma Cells

  • Lee Joo-Hyun;Han Du-Suk;Jekal Seung-Joo;Lee Jae-Hyung;Kim Chong-Ho;Yoo Min;Park Seung-Taeck
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.337-341
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    • 2005
  • This study was undertaken to clerify the cytotoxic effect of syringic acid by colorimetric assay on human cancer cells. For the evaluation of cytotoxicity of syringic acid, the cell viability and cell adhesion activity of syringic acid on cancer cells, human oral epithelioid carcinoma cells were determined using by colorimetric assays such as MTT (3-[4,5­dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and XTT (2,3-bis-[2-methoxy-4-nitro-5-sulfophenyl]­2H-tetrazolium-5-caboxanilide) assay, respectively after human oral epithelioid carcinoma cells were treated with syringic acid for 48 hours. In this study, the cell viability of syringic acid on human oral epithelioid carcinoma cells showed a significant decrease by MTT assay compared with control, and also, the cell adhesion activity by XTT assay was decreased significantly in these cells after cells were treated with various concentrations of syringic acid for 48 hours. $MTT_{50}\;and\;XTT_{50}\;were\;282.3\;{\mu}M\;and\;418.8{\mu}M$ syringic acid, respectively. These results suggest that syringic acid shows midcytotoxic effect on human oral epithelioid carcinoma cells by the decreasement of the cell viability and the cell adehision activity assessed by colorimetric assay in these cultures.

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A Colorimetric Microplate Assay Method for High Throughput Analysis of Lipase Activity

  • Choi, Suk-Jung;Hwang, Jung-Min;Kim, Sung-Il
    • BMB Reports
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    • v.36 no.4
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    • pp.417-420
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    • 2003
  • The present work describes a colorimetric microplate assay for lipase activity based on the reaction between 5,5'-dithiobis(2-nitro benzoic acid) (DTNB) and the hydrolysis product of 2,3-dimercapto-1-propanol tributyrate (DMPTB). Reaction mixtures containing DTNB, DMPTB, and lipase were prepared in microplate wells, and the absorbance at 405nm was recorded after incubation at $37^{\circ}C$ for 30 min. A linear relationship was obtained in the range of 0.1-1 U of lipase activity by this method. The reaction conditions were also optimized for the range of 0.01-0.1 U or 1-10 U. When assaying crude tissue extracts, the reaction of DTNB with non-specific reducing agents created a major source of error. However, this error was corrected by the use of blank samples that did not contain DMPTB.

Allosteric Probe-Based Colorimetric Assay for Direct Identification and Sensitive Analysis of Methicillin Resistance of Staphylococcus aureus

  • Juan Chu;Xiaoqin Zhao
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.681-688
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    • 2024
  • The accurate and rapid detection of methicillin-resistance of Staphylococcus aureus (SA) holds significant clinical importance. However, the methicillin-resistance detection strategies commonly require complicated cell lysis and gene extraction. Herein, we devised a novel colorimetric approach for the sensitive and accurate identification of methicillin-resistance of SA by combining allosteric probe-based target recognition with self-primer elongation-based target recycling. The PBP2a aptamer in the allosteric probe successfully identified the target MRSA, leading to the initiation of self-primer elongation based-cascade signal amplification. The peroxidase-like hemin/G-quadruplex undergo an isothermal autonomous process that effectively catalyzes the oxidation of ABTS2- and produces a distinct blue color, enabling the visual identification of MRSA at low concentrations. The method offers a shorter duration for bacteria cultivation compared to traditional susceptibility testing methods, as well as simplified manual procedures for gene analysis. The overall amplification time for this test is 60 min, and it has a detection limit of 3 CFU/ml. In addition, the approach has exceptional selectivity and reproducibility, demonstrating commendable performance when tested with real samples. Due to its advantages, this colorimetric assay exhibits considerable potential for integration into a sensor kit, thereby offering a viable and convenient alternative for the prompt and on-site detection of MRSA in patients with skin and soft tissue infections.