486 absorbance detector

Manufactured by Waters Corporation

Sourced in United States

The 486 Absorbance Detector is a laboratory instrument designed to measure the absorbance of light by samples. It is used to detect and quantify the concentration of specific compounds in a solution by measuring the amount of light absorbed by the sample at a specific wavelength.

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Lab products found in correlation

717 plus autosampler, by Waters Corporation (2 mentions) 0.45 m membrane, by Merck Group (2 mentions) 600 controller, by Waters Corporation (1 mentions) 2695 hplc system, by Waters Corporation (1 mentions)

Close spelling variants of this product

Waters 486 absorbance detector Waters 486 Tunable Absorbance detector 486 Tunable Absorbance Detector

4 protocols using 486 absorbance detector

HPLC-DAD Quantification of Carotenoids

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The carotenoids were quantified by HPLC-DAD, following a procedure validated by Cortés et al. [31 (link)]. The HPLC system was equipped with a 600 Controller, a 486 Absorbance Detector, a thermostatic column compartment, and a 717 Plus Auto Sampler with a cooling system (Waters, Milford, MA, USA). An aliquot of 20 µL of the extracted samples was injected and the carotenoids were separated using a reverse-phase C18 Spherisorb ODS2 (5 µm) stainless steel column (4.6 × 250 mm).
The mobile phase consisted of: (A) methanol/ammonium acetate 0.1 M, (B) milli-Q water, (C) methyl tert-butyl ether, and (D) methanol. The flow rate was fixed at 1 mL min⁻¹, and the total run time was 60 min. The column was set at 30 °C, while sample amber vials on the autosampler were preserved at 4 °C. Identification was carried out by UV–vis spectral data and their retention times [31 (link)]. Carotenoids were quantified by using calibration curves and integrating peak areas. The results are expressed on a fresh weight basis.

López-Gámez G., Elez-Martínez P., Martín-Belloso O, & Soliva-Fortuny R. (2021). Applying Pulsed Electric Fields to Whole Carrots Enhances the Bioaccessibility of Carotenoid and Phenolic Compounds in Derived Products. Foods, 10(6), 1321.

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Quantifying Carotenoids by HPLC

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Carotenoids were quantified by HPLC, following a procedure validated by Cortés et al.. [39] An aliquot of 20 µL of the extracted samples was injected into the 2695 HPLC system, which was equipped with a 600 Controller, a 486 Absorbance Detector, a thermostatic column compartment, and a 717 Plus Auto Sampler with cooling system (Waters, Milford, MA). Carotenoids were separated using a reverse-phase C18 Spherisorb ODS2 (5 µm) stainless steel column (4.6 mm x 250 mm). The mobile phase consisted of methanol/ammonium acetate 0.1 M, milli-Q water, methyl tert-butyl ether and methanol (Table 1). The flow rate was fixed at 1 mL min -1 and the total run time was 60 minutes. The column was set at 30 °C, while sample amber vials on the auto sampler were preserved at 4 °C. The carotenoids were identified by UV-vis spectral data and their retention times. [38, 39] Quantification of carotenoids was carried out by integration of the peak areas and comparison to calibration curves. Results were expressed on a fresh weight basis (mg/100 g FW).

López-Gámez G ., Elez-Martínez P ., Quiles-Chuliá A ., Martín-Belloso O ., Hernando-Hernando I , & Soliva-Fortuny R . (2021). Effect of pulsed electric fields on carotenoid and phenolic bioaccessibility and their relationship with carrot structure. Food & function, 12(6).

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HPLC-based Vitamin C Quantification

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The extraction of vitamin C (VIT C) was based on the procedure proposed by Odriozola-Serrano, Hernández-Jover and Martín-Belloso (2007). A portion of 2.5 mL of extract was added to 2.5 mL of 45 g/L metaphosphoric solution containing 1,4-Dithiothreitol (7.2 g/L). The mixture was homogenized, passed through a Millipore 0.45 µm membrane and injected in the HPLC system. The HPLC system was equipped with a 600 Controller and a 486 Absorbance Detector (Waters, Milford, MA) working at 245 nm. Samples were introduced onto the column through a manual injector equipped with a sample loop (20 ll). The flow rate was fixed at 1.0 mL/min at room temperature. A reverse-phase C18 Spherisorb® ODS2 (5 µm) stainless steel column (4.6 mm 250 mm) was used as stationary phase. The mobile phase was a 0.1 g/L solution of sulphuric acid adjusted to pH 2. 6 (Sanchez-Mata et al., 2000) . Calibration curve was built with L-ascorbic acid (0-50 mg/L). Results were expressed as mg of VIT C per 100 g of dry matter (dm).

Plazzotta S., Ibarz R., Manzocco L., & Martín‐Belloso O. (2021). Modelling the recovery of biocompounds from peach waste assisted by pulsed electric fields or thermal treatment. Journal of Food Engineering, 290, 110196-110196.

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HPLC Determination of Vitamin C

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Vitamin C content was determined based on the procedure proposed by Odriozola-Serrano, Hernández-Jover and Martín-Belloso (2007). 2.5 mL of ethanolic extract was added to 2.5 mL of 45 g/L metaphosphoric acid solution containing DTT (7.2 g/L). The mixture was homogenized, passed through a Millipore 0.45 µm membrane and injected in the HPLC system. The latter was equipped with a 600 Controller and a 486 Absorbance Detector (Waters, Milford, MA) working at 245 nm.
Samples were introduced onto the column through a manual injector equipped with a sample loop (20 ll). The flow rate was fixed at 1.0 mL/min at room temperature. A reverse-phase C18 Spherisorb® ODS2 (5 µm) stainless steel column (4.6 mm 250 mm) was used as stationary phase. The mobile phase was a 0.1 g/L solution of sulphuric acid adjusted to pH 2.6. A calibration curve was built with L-ascorbic acid (0-50 mg/L). Results were expressed as mg of vitamin C (VIT C) per 100 g dm.

Plazzotta S., Ibarz R., Manzocco L, & Martín-Belloso O. (2020). Optimizing the antioxidant biocompound recovery from peach waste extraction assisted by ultrasounds or microwaves. Ultrasonics sonochemistry, 63.

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