TP quantification was performed by UV-visible spectrophotometry according to [57 (link)]. A volume of 1 mL of diluted extract was placed in a 15 mL test tube; 6 mL of distilled water and 1 mL of Folin–Ciocalteu reagent were added, and the mixture was left to rest for 3 min. Subsequently, 2 mL of 20% Na2CO3 (w/v) was added and heated at 40 °C for 2 min. This reaction formed a blue chromophore; its absorbance was measured at 760 nm on a Shimadzu UV-VIS model 2600 spectrophotometer (Shimadzu, Kyoto, Japan). Five extraction cycles were needed to get 100% TP recovery. Results were expressed as milligrams of gallic acid equivalents (GAE) per gram of dry weight (mg GAE g−1 DW).
Uv vis model 2600 spectrophotometer
Manufactured by Shimadzu
Sourced in Japan
The Shimadzu UV-VIS model 2600 spectrophotometer is a versatile instrument designed for the measurement of absorption, transmission, and reflectance properties of liquid and solid samples. It covers a wavelength range of 185 to 900 nanometers and can be used for a variety of analytical applications in various industries.
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4 protocols using uv vis model 2600 spectrophotometer
1
Quantification of Total Phenols by UV-Vis
2
Quantification of Total Flavonoids
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TF was quantified by UV-visible spectrophotometry, according to [57 (link)]. A volume of 1 mL of the diluted extract was placed in a 15 mL tube; 4 mL of distilled water was added, and the mixture was homogenized. Then, 0.3 mL of 5% sodium nitrite (w/v) and 0.3 mL of 10% aluminum chloride (w/v) were added, and allowed the sample to stand for 5 min after the addition of each reagent. Finally, 2 mL of 1N NaOH was added, and the volume was made up to 10 mL with distilled water. This reaction formed a pink chromophore, its absorbance was measured at 490 nm with a Shimadzu UV-VIS model 2600 spectrophotometer (Shimadzu, Kyoto, Japan). Five extraction cycles were needed to reach 100% TF recovery. Results are expressed as milligrams of catechin equivalents per gram of dry sample (mg cat g−1 DW).
3
Determination of Total Carotenoids
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TC determination was performed following the method described by [58 (link)]. One gram of lyophilized sample was mixed with 1 g of calcium chloride and a solution of hexane, acetone, ethanol, and BHT in proportion 50:25:25:0.1 (v:v:v:v) to carry out the extraction. The mixture was stirred in a vortex (Mistral 4600, Multi-Mixers; Melrose, IL, USA), and centrifuged. The extract was transferred to a separatory funnel and the hexane phase was recovered in a 25 mL volumetric balloon. Finally, the extract was passed to a glass cell to read the absorbance at 490 nm on a Shimadzu UV-VIS model 2600 spectrophotometer (Kyoto, Japan). The results were expressed as micrograms of β-carotene for each gram of dry sample (DW).
4
Quantification of Total Anthocyanin Content
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TAC was calculated by UV-visible spectrophotometry, following the pH differential methodology proposed by [59 (link)]. An amount weighing 0.3 g of lyophilized sample was placed in a 15mL centrifuge tube; 5 mL of pH 1.0 buffer (KCl 0.2 N and HCl 0.2 N) was added and stirred in a Mistral Multi-Mixer vortex (Melrose Park, IL, USA) for 5 min then in a Cole Palmer model 8892 ultrasound bath (Chicago, IL, USA) and centrifuged at 5500 rpm for 10 min. The supernatant was transferred to a 25 mL amber volumetric balloon. This procedure was repeated three more times, and the mixture was brought to volume with the pH 1.0 buffer solution. Extraction with pH 4.5 buffer (CH3COONa 1M and HCl 1N) was performed using the same methodology. Quantification was performed by measuring the absorbance in the extracts (pH 1.0 and 4.5) at two wavelengths (510 and 700 nm) by a Shimadzu UV-VIS model 2600 spectrophotometer (Shimadzu, Kyoto, Japan). Seven extraction cycles were needed to reach 100% TAC recovery. Results were expressed as milligrams of cyanidin-3-glucoside chloride per gram of dry weight (mg cy-3-glu g−1 DW).
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