Agilent 8453 uv vis spectrophotometer

Manufactured by Agilent Technologies

Sourced in United States, Germany

The Agilent 8453 UV-Vis spectrophotometer is a laboratory instrument used for the quantitative analysis of compounds that absorb ultraviolet or visible light. It measures the intensity of light passing through a sample and can be used to determine the concentration of chemical species in a solution.

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Lab products found in correlation

2 6 dcpip, by Merck Group (2 mentions) Gentra puregene kit, by Qiagen (1 mentions) Quercetin, by Carl Roth (1 mentions) Pc hp 845x uv visible system, by Agilent Technologies (1 mentions) Dopamine da, by Merck Group (1 mentions) Glutathione (gsh), by Merck Group (1 mentions) Jem 2100, by JEOL (1 mentions) Gc ms, by Agilent Technologies (1 mentions) Xanthine, by Merck Group (1 mentions) X4500, by Merck Group (1 mentions) Qe6500 spectrometer, by OceanOptics (1 mentions) F3004, by Horiba (1 mentions) Fluoromax 4 spectrofluorometer, by Horiba (1 mentions) Orion star a111 benchtop ph meter, by Thermo Fisher Scientific (1 mentions) Picoharp 300, by PicoQuant (1 mentions) F 4600 spectrophotometer, by Hitachi (1 mentions) Escalab 250xi instrument, by Thermo Fisher Scientific (1 mentions) Tecnai g2, by Thermo Fisher Scientific (1 mentions) Nanoplus, by Micromeritics (1 mentions) Dialysis membrane, by Merck Group (1 mentions)

39 protocols using agilent 8453 uv vis spectrophotometer

Catalase Decomposition Activity Assay

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The activities of free catalase, co-immobilized catalase, and independently immobilized catalase were determined by following the decomposition of H₂O₂ [62 (link)]. The hybrid nanoflowers containing enzymes (5 mg) were suspended in 50 mL of deionized water and allowed to stand for 5 min. A suitable amount of catalyst sample (100 µL of 0.1 mg/mL co-immobilized catalase, 100 µL of 0.1 mg/mL independently immobilized catalase, or 100 µL of 0.01 mg/mL free catalase) was added to 10 mL of 12 mM H₂O₂ to start the decomposition reaction. The separation of co-immobilized catalase and independently immobilized catalase from the reaction media was carried out by the filtration steps described above. The absorbance decrease of filtrate at 240 nm (ε = 43.6 M⁻¹ cm⁻¹) was monitored using the Agilent 8453 UV-VIS spectrophotometer. The determination of the activity of free enzymes followed the same procedure, but without the filtration step. One unit was defined as the amount of enzyme that decomposed 1.0 µ mol of H₂O₂ per minute per milligram of catalase encapsulated into the hybrid nanoflowers, or free catalase under the conditions described.

Wu Z., Shi L., Yu X., Zhang S, & Chen G. (2019). Co-Immobilization of Tri-Enzymes for the Conversion of Hydroxymethylfurfural to 2,5-Diformylfuran. Molecules, 24(20), 3648.

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DNA Adduct Analysis via LC-MS

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Cell pellets (1.5 – 2 × 10⁶) were homogenized in TE buffer (300 μL, 50 mM Tris-HCl containing 10 mM EDTA, and 10 mM BME, pH 8.0) and DNA was purified with the Gentra Puregene kit (Qiagen, Valencia, CA) (Cai et al. 2017 (link)). The DNA was washed with 70%, ethanol and reconstituted in LC/MS grade water. The concentration of DNA was determined using an Agilent 8453 UV/vis spectrophotometer (Agilent Technologies, Santa Clara, CA). Ten μg of DNA were spiked with isotopically labeled internal standards ([¹³C₁₀]-dG-C8–4-ABP, [¹³C₁₀]-dG-C8-PhIP, [¹³C₁₀]-dG-C8-AαC, [²H₃C]-dG-C8-MeIQx, each at a level of 1 adducts per 10⁷ nucleotides) and were digested with a cocktail of enzymes in 5 mM Bis-Tris-HCl buffer (pH 7.1) as previously described (Goodenough et al. 2007 (link); Nauwelaers et al. 2011 (link)). Isotopically labeled 2-NA DNA adducts were not available, and [¹³C₁₀]-dG-C8–4-ABP was used as a surrogate internal standard assuming similar ionization efficiencies for all adducts.

Bellamri M., Yao L., Bonala R., Johnson F., Von Weymarn L.B, & Turesky R.J. (2019). Bioactivation of the tobacco carcinogens 4-aminobiphenyl (4-ABP) and 2-amino-9H-pyrido[2,3-b]indole (AαC) in human bladder RT4 cells. Archives of toxicology, 93(7), 1893-1902.

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Comparative Enzyme Assay of Immobilized and Free HRP

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The activities of free horseradish peroxidase, co-immobilized horseradish peroxidase, and independently immobilized horseradish peroxidase were assayed using pyrogallol and H₂O₂ as substrates [63 ]. The reaction mixture contained: 100 mM phosphate buffer, pH 6.0, 10 mM H₂O₂ and 13 mM pyrogallol, and a suitable amount of catalyst sample (100 µL of 0.1 mg/mL co-immobilized horseradish peroxidase, 100 µL of 0.1 mg/mL independently immobilized horseradish peroxidase or 100 µL of 0.01 mg/mL free horseradish peroxidase) in a final volume of 20 mL. The reaction mixture was incubated at 30 °C for 3 min. The absorbance increase at 430 nm (ε = 2.47 mM^−l cm⁻¹) was detected using the Agilent 8453 UV-VIS spectrophotometer. As mentioned above, co-immobilized horseradish peroxidase and independently immobilized horseradish peroxidase were separated by a filtration step to complete the UV-VIS detection. One unit was defined as the amount of enzyme that produced 1.0 µ mol of purpurogallin per minute per milligram of horseradish peroxidase encapsulated into the hybrid nanoflowers, or free horseradish peroxidase.

Wu Z., Shi L., Yu X., Zhang S, & Chen G. (2019). Co-Immobilization of Tri-Enzymes for the Conversion of Hydroxymethylfurfural to 2,5-Diformylfuran. Molecules, 24(20), 3648.

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Quantifying β-Carotene Encapsulation in SLNs

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Starting with a stock solution of pure β-carotene in n-hexane, sequential dilutions were made in duplicate, and the absorbance at 450 nm was read in triplicate using an Agilent 8453 UV-Vis Spectrophotometer (Agilent Technologies, Waldbronn, Germany). The calibration curve was linear, with a correlation coefficient R² of 0.99227, over a concentration range of 0.5–5.0 µg/mL. The overall amount of β-carotene loaded in the suspension was determined by drying 200 µL of SLN and solubilizing the residue in a known volume of n-hexane (10 mL). Then, solutions were analyzed using a UV-Vis spectrophotometer to determine the β-carotene concentration. Finally, the encapsulation efficiency (EE) of SLN was determined using the following equation:

% E E = \frac{β - carotene quantified in the SLN}{β - carotene initially added to the lipid} \times 100

Pinna N., Ianni F., Blasi F., Stefani A., Codini M., Sabatini S., Schoubben A, & Cossignani L. (2022). Unconventional Extraction of Total Non-Polar Carotenoids from Pumpkin Pulp and Their Nanoencapsulation. Molecules, 27(23), 8240.

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Soxhlet Extraction and UV-Vis Polyphenol Analysis

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A Soxhlet apparatus was used for drug extraction. The quantitative analysis of the polyphenolic substances was performed with an Agilent 8453 UV/Vis spectrophotometer (Agilent, Germany) equipped with PC -HP 845x UV—Visible System (Agilent, Germany) and 1 cm quartz cuvettes.
With the exception of the Folin–Ciocalteu phenol reagent (FCR), casein (Merck, Darmstadt, Germany) and quercetin (Roth, Karlsruhe, Germany) all chemicals and reagents for polyphenol analysis were of analytical quality grade and were supplied by Kemika (Zagreb, Croatia).

Nazlić M., Kremer D., Grubešić R.J., Soldo B., Vuko E., Stabentheiner E., Ballian D., Bogunić F, & Dunkić V. (2020). Endemic Veronica saturejoides Vis. ssp. saturejoides–Chemical Composition and Antioxidant Activity of Free Volatile Compounds. Plants, 9(12), 1646.

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Kinetics of NahK Decarboxylation Reaction

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The NahK decarboxylation activity at 25 °C was monitored by following the rate of appearance of the product 2-hydroxy-2,4-pentadienoate at 265 nm (ε = 7,850 M⁻¹ cm⁻¹^{(12 (link))}) in an Agilent 8453 UV-Vis spectrophotometer (Agilent Technologies). The reaction mixtures consisted of 20 mM sodium phosphate buffer in the pH range 6–8, 5 mM MgCl₂, and 0.5 µg/mL of NahK. The reaction was initiated by addition of the substrate (1.6–48.0 µM) in a final reaction volume of 1.0 mL. The stock solution of substrate (2-oxo-3-hexenedioate) was generated enzymatically by the action of 4-oxalocrotonate tautomerase (XylH ^{(6 )}) on 2-hydroxymuconate. The 2-hydroxymuconate was previously dissolved in ethanol then made up in 20 mM sodium phosphate buffer pH 7.4. The V_max and K_M values, and their standard errors, were estimated by non-linear regression of the initial rate of the reactions versus the concentration of the substrate to the Michaelis-Menten equation using the software GraFit (Erathicus Software Ltd., Staines, U.K.).

Guimarães S.L., Coitinho J.B., Costa D.M., Araújo S.S., Whitman C.P, & Nagem R.A. (2016). Crystal structures of apo and liganded 4-oxalocrotonate decarboxylase uncover a structural basis for the metal-assisted decarboxylation of a vinylogous β-keto acid. Biochemistry, 55(18), 2632-2645.

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Platinum@Silver Nanoflowers Synthesis and Characterization

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Silver nitrate (AgNO₃), chloroplatinic acid (H₂PtCl₆·6H₂O), bovine serum albumin (BSA), ascorbic acid (AA), 3,3′,5,5′-tetramethylbenzidine (TMB), cysteine (Cys) and glucose (Glu) were purchased in Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).
Uric acid (UA), uricase, dopamine (DA), and glutathione (GSH) were obtained from Sigma-Aldrich (Shanghai, China). Urea, hydrogen peroxide (30 wt% H₂O₂), KNO₃, Mg (NO₃)₂, NaCl, CaCl₂ were obtained from Beijing Chemical Industry Co., Ltd. (Beijing, China). All reagents are analytical grade and do not require further treatment. Experimental water was ultrapure water (18.25 MΩ, Millipore, USA).
UV-vis spectral measurements were recorded on Agilent 8453 UV-vis spectrophotometer (Agilent Technologies, Santa Clara, CA, USA). The morphology of the Pt@Ag NFs were characterized by transmission electron microscopy (TEM, JEM-2100, JEOL Co., Japan). X-ray photoelectron spectroscopy (XPS) was carried out on EscaLab 250Xi with Al K α X-ray radiation as the source for excitation. Biochemical analysis was measured using biochemical analyzer (AU2007, Beckman, Olympus, USA).

Wang X., Chen S., Tang X., Lin D, & Qiu P. (2019). Ultrasensitive detection of uric acid in serum of patients with gout by a new assay based on Pt@Ag nanoflowers. RSC Advances, 9(63), 36578-36585.

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Spectrophotometric and HPLC-based Assays

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Individual assays were measured spectrophotometrically using an Agilent 8453 UV-Vis spectrophotometer (Agilent Technologies). Organic acids were measured by HPLC (Agilent Technologies and Thermo Fisher Scientific). Labeled acetate and formate were analyzed by GC-MS (Agilent Technologies). Details are provided in SI Appendix.

Lin P.P., Jaeger A.J., Wu T.Y., Xu S.C., Lee A.S., Gao F., Chen P.W, & Liao J.C. (2018). Construction and evolution of an Escherichia coli strain relying on nonoxidative glycolysis for sugar catabolism. Proceedings of the National Academy of Sciences of the United States of America, 115(14), 3538-3546.

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Quantifying HDL Oxidation Kinetics

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HDL (0.01 mg/mL cholesterol, final) was incubated with XO (14.3 µU/mL, final, Sigma-Aldrich, X4500, St. Louis, MO), xanthine (0.02 µM, final, Sigma-Aldrich, X7379, St. Louis, MO), and Hb (0, 4 and 16 mg/dL, final, Sigma-Aldrich, X7375) at room temperature for 100 min, during which A_{234 nm} was recorded by Agilent 8453 UV-Vis spectrophotometer (Agilent Technologies, Santa Clara, CA). The initial rate of HDL conjugated diene formation/oxidation was expressed as the slope of linear regression of A_{234 nm} over the first 40 min.

Ji X., Xu H., Zhang H., Hillery C.A., Gao H.Q, & Pritchard KA J.r. (2014). Anion Exchange HPLC Isolation of High-Density Lipoprotein (HDL) and On-Line Estimation of Proinflammatory HDL. PLoS ONE, 9(3), e91089.

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Characterizing Absorption and Fluorescence of Organic Dimers

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Absorption spectra were measured using an Agilent 8453 UV/Vis spectrophotometer. A Fluoromax‐4 spectrofluorometer (Jobin–Yvon, Horiba) was used for measuring fluorescence spectra of dimers 2, 3 and 4, fluorescence spectra of dimer 1 were measured by an Ocean Optics QE 6500 spectrometer. The temperature of cuvette in the fluorometer was controlled by a Peltier thermostated cuvette holder (F3004, Jobin–Yvon, Horiba) to 0.5 °C accuracy. Quartz cuvettes with 10 mm path length were used. The concentration of the dimers for absorption and fluorescence measurements was 0.33 μm leading to absorbance of ≈0.1.

Vyšniauskas A., Ding D., Qurashi M., Boczarow I., Balaz M., Anderson H.L, & Kuimova M.K. (2017). Tuning the Sensitivity of Fluorescent Porphyrin Dimers to Viscosity and Temperature. Chemistry (Weinheim an Der Bergstrasse, Germany), 23(46), 11001-11010.

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