Lambda 12 spectrophotometer

Manufactured by PerkinElmer

Sourced in United States, Canada

The Lambda 12 spectrophotometer is an analytical instrument designed for UV-Vis spectroscopy. It measures the absorption or transmittance of light by a sample over a range of wavelengths, providing data on the sample's composition and properties.

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Ilab 600, by Werfen (1 mentions) Capillary melting point apparatus, by Büchi (1 mentions)

7 protocols using lambda 12 spectrophotometer

Antioxidant Capacity Evaluation of Maize

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Antioxidant capacity assays were carried out on the same extracts used for phenolic profiling. The in vitro antioxidant capacity of each maize sample was evaluated as radical scavenging ability (DPPH assay) and ferric reducing power (FRAP assay), as previously described (Ghisoni et al., 2017 (link)). Briefly, 1.5 mL of maize extract was combined to the same volume of an ethanol solution of DPPH (1.0 mM). The absorbance was recorded at 5-min intervals (until the steady state) to 517 nm using a Perkin Elmer Lambda 12 spectrophotometer (Ontario, Canada). The results were finally expressed as gallic acid equivalents (GAE).
The FRAP antioxidant assay was carried out by means of a clinical analyzer ILAB 600 (Instrumentation Laboratory, Lexington, MA, United States). The FRAP working reagent consist of acetate buffer (300 mM, pH 3.6), TPTZ (10 mM) in 40 mM HCl and FeCl3 (20 mM), in the ratio 10:1:1 (v/v). Each extract (100 μL) was combined to 3 ml of FRAP working reagent, and the absorbance was recorded at λ = 600 nm, after 243 s of incubation at 37°C. The FRAP results were expressed as GAE.

Bernardi J., Stagnati L., Lucini L., Rocchetti G., Lanubile A., Cortellini C., De Poli G., Busconi M, & Marocco A. (2018). Phenolic Profile and Susceptibility to Fusarium Infection of Pigmented Maize Cultivars. Frontiers in Plant Science, 9, 1189.

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Quantifying Astaxanthin Content via Spectrophotometry

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Spectrophotometric measurements were set up on a Lambda 12 Spectrophotometer (PerkinElmer Inc., Norwalk, CT, USA) in the UV-visible domain (200–800 nm), using a scan speed of 50 nm/min. A calibration curve based on λ_max absorption was obtained using AstaN (0–50 μM) in EtOH/DCM (% v/v; 60/40). The extinction coefficient (ε) was used to calculate the astaxanthin content in AstaCO₂ and AstaCO₂-NLC.

Rodriguez-Ruiz V., Salatti-Dorado J.Á., Barzegari A., Nicolas-Boluda A., Houaoui A., Caballo C., Caballero-Casero N., Sicilia D., Bastias Venegas J., Pauthe E., Omidi Y., Letourneur D., Rubio S., Gueguen V, & Pavon-Djavid G. (2018). Astaxanthin-Loaded Nanostructured Lipid Carriers for Preservation of Antioxidant Activity. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(10), 2601.

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Quantifying Lipid Peroxidation via TBARS Assay

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We performed the TBARS assay to quantify the lipid peroxidation marker malondialdehyde (MDA), according to the method reported by Zhang and Huang [16 ], with minor modifications. Briefly, 10 mL of cell culture was centrifuged for 10 min at 10,000× g. The pellet was then mixed in a tube with 1 mL 0.1% trichloroacetic acid (TCA), and placed in an ultra-sonic bath at room temperature for 3 min to promote extraction. Next, the tube was centrifuged (10,000× g for 10 min at 4 °C) and the supernatant was transferred to a new tube. A 200 µL aliquot of supernatant was mixed well with 1 mL 20% TCA containing 0.5% TBA. This mixture was boiled at 95 °C for 15 min, quickly cooled on ice, centrifuged at 10,000× g for 5 min, and then the supernatant was transferred to a cuvette. Finally, we measured the absorbance at 532 nm using a PerkinElmer Lambda 12 spectrophotometer (PerkinElmer, Shelton, CT, USA). A molar extinction coefficient of 155 cm⁻¹ mM⁻¹ was used for MDA determination. The results were corrected for the experimental blank, and the MDA concentration was calculated as described by Lin et al. [17 (link)].

Senizza A., Callegari M.L., Senizza B., Minuti A., Rocchetti G., Morelli L, & Patrone V. (2019). Effects of Linoleic Acid on Gut-Derived Bifidobacterium breve DSM 20213: A Transcriptomic Approach. Microorganisms, 7(12), 710.

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Colorimetric Assay for Fe2+ Chelation by LA

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We assessed the ability of LA to chelate Fe²⁺ using the colorimetric assay previously reported by Carter [18 (link)], with minor modifications. Briefly, 1.61 mL of each sample (MRS-cys) was diluted 1:30 (v/v) and then transferred into a 2-mL cuvette, with the addition of 150 µL 0.30 mmol/L FeSO₄ and 0.5 g/L LA. After a 5-min reaction time, we added 230 µL 0.80 mmol/L ferrozine solution to each cuvette. In the control, the ferrozine solution was replaced by distilled water to correct for the unequal colors of the sample solutions. After 15 min, we read the absorbance at 562 nm using a PerkinElmer Lambda 12 spectrophotometer (PerkinElmer, Shelton, CT, USA). The final results were expressed as mg EDTA equivalents/L, according to Santos and co-authors [19 (link)].

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Detailed Synthesis of Pyrylium Dyes

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All reagents were purchased from commercial sources and, unless otherwise stated, were used without further purification. Reactions run under anhydrous and/or inert conditions are noted in the procedure as such. All other reactions were performed exposed to atmospheric conditions. Tetrahydrofuran (THF) was distilled from a sodium benzophenone ketyl, and other anhydrous solvents were dried over 4 Å molecular sieves before use. Concentration in vacuo was performed on a rotary evaporator. NMR spectra were recorded on either a 300 MHz Gemini or a 500 MHz Varian Inova spectrometer for ¹H NMR spectroscopy and a 300 MHz Gemini spectrometer operating at 75.5 MHz for ¹³C NMR spectroscopy. Residual solvent signal was used as the internal standard. If a mixture of CD₂Cl₂ and CD₃OD was used, the residual peak for CH₂Cl₂ was used as the internal standard. UV/VIS-near-IR spectra were recorded in quartz cuvettes with a 1-cm path length on a Perkin Elmer Lambda 12 spectrophotometer. Melting points were determined with a Büchi capillary melting point apparatus and are uncorrected. ¹³C NMR spectra were not recorded for some pyrylium dyes due to limited solubility in common NMR solvents. Detailed synthesis for each is presented in the SI.

Misra V.K, & Draper D.E. (2001). A thermodynamic framework for Mg2+ binding to RNA. Proceedings of the National Academy of Sciences of the United States of America, 98(22), 12456-12461.

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Measuring MDA Levels in Cell Cultures

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The MDA content was measured at lag, log and stationary phases by using the thiobarbituric acid reactive substance (TBARS) assay, as previously reported^{46 (link)}. Pellets were obtained from 10 mL of cell culture by centrifuging for 10 min at 10,000 × g. The absorbance at 532 nm was determined using a Perkin Elmer (Ontario, Canada) Lambda 12 spectrophotometer. For MDA determination, a molar extinction coefficient of 155 cm⁻¹ mM⁻¹ was used. Results were finally expressed as nM MDA equivalents (n = 3).

Senizza A., Rocchetti G., Callegari M.L., Lucini L, & Morelli L. (2020). Linoleic acid induces metabolic stress in the intestinal microorganism Bifidobacterium breve DSM 20213. Scientific Reports, 10, 5997.

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Quantification of Phenolic Compounds

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The extractable phenolic compounds, hydrolyzable phenolic compounds and nonextractable proanthocyanidins fractions were used for spectrophotometric determinations of total phenolic compounds content (corresponding to the sum of the three fractions). Briefly, the Folin-Ciocalteu assay 24 was applied to the extractable and hydrolyzable phenolic compound fractions; the results were expressed as g of gallic acid equivalents/100 g dw.
The non-extractable proanthocyanidin content was determined in the hydrolyzates by measuring the sum of absorbance at 450 and 555 nm; the results were expressed as mg of non-extractable proanthocyanidins/100 g dw, by using a standard curve from a polymeric proanthocyanidin concentrate. 25 All these measurements were carried out in a Lambda 12 spectrophotometer (Perkin-Elmer; Waltham, MA, U.S.A.).

Pérez-Ramírez I.F., Reynoso-Camacho R., Saura-Calixto F, & Pérez-Jiménez J. (2018). Comprehensive Characterization of Extractable and Nonextractable Phenolic Compounds by High-Performance Liquid Chromatography-Electrospray Ionization-Quadrupole Time-of-Flight of a Grape/Pomegranate Pomace Dietary Supplement. Journal of agricultural and food chemistry, 66(3).

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