The TPC of SCG extracts was determined by a Folin-Ciocalteu assay [33 ] (Spectrometer UV 1240 Shimadzu, Kioto, Japan). Results were expressed as g of gallic acid equivalent (GAE) per g of dry spent coffee.
Spectrometer uv 1240
Manufactured by Shimadzu
Sourced in Japan
The Spectrometer UV 1240 is a single-beam ultraviolet-visible (UV-VIS) spectrophotometer designed for general laboratory analysis. It has a wavelength range of 190 to 1100 nanometers and provides absorption and transmittance measurements.
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7 protocols using spectrometer uv 1240
1
Folin-Ciocalteu Assay for SCG Extract
2
Antioxidant Capacity of Spent Coffee Grounds
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The antioxidant capacity of SCG extracts was determined by DPPH and an improved version of the ORAC assay developed by Ou et al. [34 (link)].
DPPH was performed by spectrophotometry (Spectrometer UV 1240, Shimadzu, Kioto, Japan) according to the free radical 2,2-diphenyl-1-picrylhydrazyl method (DPPH) [35 ]. The efficient concentration of extracts, which is the concentration necessary to inhibit 50% the absorption of DPPH (EC50; mg/mL), was determined in triplicate and expressed as mg of Trolox equivalents per g of dry SCG (mg TE/g).
The ORAC assay was carried out in a PerkinElmer 2030 Multilabel Reader with 96-well black plates [34 (link)]. The Trolox equivalent molar concentrations of the samples were calculated using a linear regression equation between the Trolox concentration and the corresponding net AUC [34 (link)]. To compare the antioxidant activity of the extracts, we decided to calculate the relative ORAC values as mg of Trolox equivalents present in 1 g of dry SCG.
DPPH was performed by spectrophotometry (Spectrometer UV 1240, Shimadzu, Kioto, Japan) according to the free radical 2,2-diphenyl-1-picrylhydrazyl method (DPPH) [35 ]. The efficient concentration of extracts, which is the concentration necessary to inhibit 50% the absorption of DPPH (EC50; mg/mL), was determined in triplicate and expressed as mg of Trolox equivalents per g of dry SCG (mg TE/g).
The ORAC assay was carried out in a PerkinElmer 2030 Multilabel Reader with 96-well black plates [34 (link)]. The Trolox equivalent molar concentrations of the samples were calculated using a linear regression equation between the Trolox concentration and the corresponding net AUC [34 (link)]. To compare the antioxidant activity of the extracts, we decided to calculate the relative ORAC values as mg of Trolox equivalents present in 1 g of dry SCG.
3
Antioxidant Capacity of Red Tara Extract
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The AOC of red Tara pods extract was determined by spectrophotometry (Spectrometer UV 1240, Shimadzu, Kioto, Japan) according to the free radical 2,2-diphenyl- 1-picrylhydrazyl method (DPPH) and expressed as the efficient concentration of extract (EC50: mg/mL), which is the concentration necessary to inhibit 50% of radical absorption of DPPH [23 (link)].
4
Quantifying Tara Pods Polyphenols
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The TPC of red Tara pods extract was determined by the spectrophotometric Folin−Ciocalteu assay (Spectrometer UV 1240, Shimadzu, Kioto, Japan) and expressed as g of gallic acid equivalent (GAE) per g of dry Tara pods [22 ].
5
Antioxidant Capacity Determination of Plant Extracts
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The antioxidant capacity of the extracts was determined using the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging method [50 ]. In summary, first 0.1 mL of extract was mixed with 3.9 mL of DPPH solution (0.1 mM), then the solution was mixed by vortex for 10 s and incubated at room temperature in the dark for 30 min. The reduction of DPPH was measured at 517nm (Spectrometer UV 1240, Shimadzu, Kioto, Japan). The IC50 (mg/L), defined as the effective extract concentration needed to inhibit 50% of DPPH radical absorption, was calculated and compared with Trolox, using the Trolox equivalent antioxidant capacity (TEAC) equation (TEAC = IC50 Trolox/IC50 sample) [51 (link)]. Antioxidant capacity values were expressed as µM of Trolox equivalent (TE) per gram of dry mass of pomace.
6
Polyphenol Content in Carménère Pomace
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Total polyphenols content (TPC) levels of the Carménère pomace extracts were determined by Folin–Ciocalteu assay [49 ]. A volume of 3.75 mL of distilled water, 0.5 mL of Carménère pomace extract and 0.25 mL of Folin–Ciocalteu reactive (1N) were mixed with 0.5 mL of a sodium carbonate solution (10% w/v). Absorbance was measured at 765 nm (Spectrometer UV 1240, Shimadzu, Kioto, Japan) after a reaction time of 1 h at 20 °C. Results were expressed as mg of GAE per gram of dried pomace.
7
Antioxidant Capacity Evaluation of Pomace
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The antioxidant capacity of the extracts was determined using spectrophotometry (Spectrometer UV 1240, Shimadzu, Kyoto, Japan) at 517 nm with the DPPH radical scavenging method [31 (link)]. In summary, 0.1 mL of diluted extract and 3.9 mL of DPPH solution (0.1 mM) were mixed, and then the mixture was protected from light for 30 min at room temperature. Two controls were used for this assay; (i) 3.9 mL of methanol (dilution blank) and (ii) 3.9 mL of the methanolic solution of DPPH (negative control). The effective extract concentration required to inhibit 50% of the DPPH radical absorption (IC50; mg/L) was calculated. The extract antioxidant capacity was compared with Trolox using the Trolox equivalent antioxidant capacity (TEAC) equation: TEAC = IC50 Trolox/IC50 sample. DPPH values were expressed as µmol of Trolox equivalent (TE) per gram of dry mass of pomace (µM TE/gdp).
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