A Simple and a Reliable Method to Quantify Antioxidant Activity In Vivo
Abstract
:1. Introduction
2. Material and Methods
2.1. Preliminary Assay
2.2. Green Tea Extract Preparation
2.3. Antioxidant Activity In Vitro
2.4. Antioxidant Activity In Vivo
2.4.1. Worms and Plate Preparation
2.4.2. DPPH• Assay
2.4.3. Limitations and Precautions of the Method
2.5. Statistical Analysis
3. Results
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Sample | Time (min) | |||||||
---|---|---|---|---|---|---|---|---|
Concentration (µg/mL) | 0 | 15 | 30 | 45 | 60 | 75 | 90 | |
Green tea | 500 | 97.42 ± 0.64 | 97.45 ± 0.62 | 97.09 ± 0.57 | 96.46 ± 0.55 | 96.17 ± 0.61 | 95.30 ± 1.06 | 93.29 ± 1.44 |
250 | 26.54 ± 0.39 | 96.97 ± 0.42 | 96.63 ± 0.41 | 96.29 ± 0.41 | 95.87 ± 0.50 | 95.43 ± 0.47 | 96.64 ± 0.77 | |
125 | 96.13 ± 0.70 | 96.83 ± 0.68 | 96.52 ± 1.19 | 95.88 ± 1.18 | 95.51 ± 0.72 | 94.98 ± 0.56 | 94.40 ± 0.99 | |
62.50 | 93.71 ± 0.59 | 95.14 ± 0.51 | 94.75 ± 0.49 | 894.01 ± 0.57 | 93.38 ± 0.44 | 93.02 ± 0.36 | 92.44 ± 0.43 | |
31.25 | 90.50 ± 1.17 | 94.50 ± 0.38 | 94.31 ± 0.17 | 93.60 ± 0.49 | 92.82 ± 0.52 | 92.27 ± 056 | 91.62 ± 0.61 | |
15.62 | 84.57 ± 1.25 | 93.20 ± 0.47 | 93.37 ± 0.44 | 92.86 ± 0.38 | 92.06 ± 0.51 | 91.06 ± 0.49 | 90.43 ± 0.53 | |
7.81 | 71.69 ± 1.91 | 86.90 ± 1.06 | 88.26 ± 0.92 | 88.52 ± 1.02 | 88.27 ± 0.89 | 88.13 ± 0.81 | 87.84 ± 0.87 | |
3.91 | 44.40 ± 0.40 | 59.48 ± 1.27 | 62.64 ± 1.32 | 63.88 ± 1.51 | 64.43 ± 1.47 | 64.84 ± 1.59 | 64.82 ± 1.58 | |
1.95 | 26.48 ± 0.54 | 35.60 ± 0.74 | 36.78 ± 1.28 | 37.07 ± 0.95 | 36.69 ± 1.25 | 36.00 ± 1.52 | 34.75 ± 1.30 | |
0.98 | 16.97 ± 1.01 | 21.14 ± 0.96 | 21.08 ± 1.42 | 20.66 ± 1.37 | 19.83 ± 1.29 | 19.07 ± 1.36 | 17.40 ± 1.53 | |
RA | 15.62 | 100.00 ± 3.69 | 94.09 ± 1.48 | 93.31 ± 9.69 | 93.87 ± 1.46 | 93.53 ± 1.29 | 92.97 ± 1.40 | 92.23 ± 1.63 |
7.81 | 97.80 ± 2.98 | 91.94 ± 1.67 | 91.99 ± 1.71 | 92.12 ± 2.53 | 91.89 ± 1.33 | 91.57 ± 2.29 | 91.06 ± 2.63 | |
3.91 | 82.66 ± 3.80 | 84.59 ± 1.72 | 86.49 ± 9.72 | 87.84 ± 1.25 | 88.78 ± 2.03 | 89.46 ± 1.68 | 89.60 ± 1.40 | |
1.95 | 44.59 ± 3.37 | 47.77 ± 1.12 | 50.05 ± 8.20 | 52.25 ± 1.25 | 54.08 ± 1.29 | 54.81 ± 1.52 | 54.84 ± 1.60 | |
0.98 | 18.88 ± 1.85 | 18.93 ± 1.30 | 20.68 ± 1.25 | 21.63 ± 1.24 | 22.05 ± 1.40 | 20.98 ± 1.16 | 18.51 ± 1.72 | |
Gluc. | 500 | −0.70 ± 1.68 | −0.72 ± 7.52 | 0.09 ± 7.46 | 0.80 ± 7.40 | 0.62 ± 7.52 | 0.40 ± 6.89 | 1.97 ± 6.79 |
250 | 1.34 ± 4.64 | −1.16 ± 5.39 | −1.19 ± 5.84 | −1.65 ± 17.41 | −2.59 ± 16.64 | −3.49 ± 18.20 | −3.05 ± 19.15 | |
125 | 0.38 ± 6.53 | −4.18 ± 1.38 | −0.48 ± 2.48 | −11.67 ± 2.14 | −13.21 ± 2.40 | −15.00 ± 2.87 | −15.30 ± 2.54 | |
62.50 | −0.67 ± 5.96 | 1.57 ± 9.11 | −6.70 ± 6.51 | −8.06 ± 16.67 | −9.68 ± 17.28 | −11.52 ± 8.37 | −12.01 ± 8.67 | |
31.25 | 4.51 ± 5.03 | −6.62 ± 4.69 | −8.18 ± 5.06 | −9.49 ± 6.03 | −11.57 ± 5.47 | −14.00 ± 6.78 | −14.62 ± 6.83 | |
15.62 | 1.07 ± 6.21 | 2.52 ± 11.54 | 1.76 ± 11.97 | 0.17 ± 12.40 | −1.86 ± 12.05 | −3.90 ± 13.02 | −4.83 ± 13.42 | |
7.81 | 2.62 ± 3.85 | 2.93 ± 4.57 | 1.49 ± 5.18 | −0.10 ± 5.91 | −2.10 ± 5.26 | −4.11 ± 6.49 | −4.91 ± 6.85 | |
3.91 | −4.68 ± 7.57 | −2.43 ± 1.19 | −2.27 ± 1.30 | −2.40 ± 2.86 | −2.16 ± 2.00 | −2.98 ± 2.75 | −3.31 ± 2.26 | |
1.95 | −1.09 ± 5.32 | −6.39 ± 6.30 | −7.42 ± 7.11 | −8.72 ± 8.09 | −10.88 ± 7.54 | −13.26 ± 8.75 | −14.03 ± 9.23 | |
0.98 | −2.13 ± 7.00 | −7.46 ± 7.58 | −8.10 ± 7.98 | −9.30 ± 8.53 | −11.00 ± 8.53 | −12.76 ± 9.08 | −12.98 ± 8.82 |
Sample | Time (min) | |||||||
---|---|---|---|---|---|---|---|---|
Concentration (µg/mL) | 0 | 15 | 30 | 45 | 60 | 75 | 90 | |
Green tea | 250 | 86.25 ± 0.50 | 87.24 ± 0.76 | 87.80 ± 0.41 | 88.42 ± 0.47 | 88.88 ± 0.66 | 89.07 ± 0.41 | 89.89 ± 0.52 |
100 | 55.91 ± 0.90 | 75.27 ± 0.86 | 79.91 ± 1.10 | 82.98 ± 1.52 | 86.10 ± 1.43 | 87.17 ± 1.74 | 88.19 ± 1.45 | |
50 | 27.46 ± 0.95 | 47.49 ± 0.87 | 54.37 ± 1.58 | 57.77 ± 1.73 | 61.84 ± 0.94 | 63.74 ± 1.10 | 66.78 ± 1.11 | |
5 | 16.55 ± 0.95 | 17.17 ± 0.95 | 23.11 ± 0.85 | 26.31 ± 0.81 | 28.20 ± 0.89 | 30.34 ± 1.10 | 32.54 ± 0.77 | |
0.5 | 11.99 ± 0.83 | 13.74 ± 0.83 | 18.41 ± 0.91 | 21.34 ± 1.41 | 24.78 ± 0.55 | 27.47 ± 0.75 | 29.06 ± 0.74 | |
0 | 1.32 ± 1.10 | 2.13 ± 1.19 | 2.35 ± 1.42 | 3.18 ± 1.32 | 3.53 ± 1.27 | 4.04 ± 1.23 | 4.51 ± 1.24 | |
RA | 250 | 87.29 ± 0.94 | 87.72 ± 0.53 | 88.32 ± 0.57 | 88.92 ± 0.65 | 89.37 ± 0.73 | 89.59 ± 0.88 | 89.72 ± 0.56 |
100 | 85.60 ± 0.87 | 86.13 ± 0.65 | 86.12 ± 0.62 | 86.31 ± 0.85 | 87.11 ± 0.83 | 87.18 ± 0.71 | 87.24 ± 0.76 | |
50 | 83.65 ± 4.24 | 85.72 ± 0.38 | 86.27 ± 0.33 | 86.38 ± 0.33 | 86.56 ± 0.40 | 86.66 ± 0.50 | 86.96 ± 0.42 | |
5 | 14.88 ± 0.60 | 31.30 ± 0.83 | 34.47 ± 0.80 | 37.02 ± 0.60 | 38.71 ± 0.45 | 38.70 ± 0.73 | 38.98 ± 0.65 | |
0.5 | 4.23 ± 0.52 | 18.79 ± 0.62 | 21.53 ± 0.51 | 23.09 ± 0.55 | 25.08 ± 1.11 | 25.10 ± 0.51 | 25.17 ± 0.54 | |
0 | 1.93 ± 0.86 | 2.53 ± 0.86 | 2.64 ± 0.89 | 2.95 ± 1.00 | 3.51 ± 0.74 | 4.15 ± 0.83 | 5.10 ± 0.82 | |
Glucose | 250 | 12.15 ± 0.74 | 21.73 ± 0.66 | 22.98 ± 0.83 | 23.41 ± 0.95 | 24.03 ± 1.17 | 25.42 ± 1.43 | 26.15 ± 1.79 |
100 | 9.47 ± 0.80 | 10.67 ± 2.82 | 11.73 ± 2.49 | 11.95 ± 2.04 | 13.41 ± 1.89 | 13.54 ± 1.75 | 14.92 ± 1.51 | |
50 | 6.05 ± 0.47 | 8.30 ± 0.80 | 10.29 ± 0.82 | 11.08 ± 0.88 | 11.28 ± 0.96 | 12.20 ± 1.21 | 13.16 ± 1.19 | |
5 | 4.49 ± 0.79 | 6.95 ± 0.89 | 7.13 ± 0.82 | 7.44 ± 0.62 | 7.26 ± 0.45 | 8.38 ± 0.58 | 9.41 ± 0.62 | |
0.5 | 3.66 ± 1.00 | 4.36 ± 0.86 | 5.38 ± 0.87 | 5.28 ± 0.81 | 5.60 ± 0.82 | 6.49 ± 0.79 | 6.62 ± 0.93 | |
0 | 1.76 ± 0.95 | 2.07 ± 0.97 | 2.85 ± 0.68 | 3.06 ± 1.30 | 3.40 ± 1.24 | 3.83 ± 1.30 | 4.67 ± 1.26 |
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de Torre, M.P.; Cavero, R.Y.; Calvo, M.I.; Vizmanos, J.L. A Simple and a Reliable Method to Quantify Antioxidant Activity In Vivo. Antioxidants 2019, 8, 142. https://doi.org/10.3390/antiox8050142
de Torre MP, Cavero RY, Calvo MI, Vizmanos JL. A Simple and a Reliable Method to Quantify Antioxidant Activity In Vivo. Antioxidants. 2019; 8(5):142. https://doi.org/10.3390/antiox8050142
Chicago/Turabian Stylede Torre, María Pilar, Rita Yolanda Cavero, María Isabel Calvo, and José Luis Vizmanos. 2019. "A Simple and a Reliable Method to Quantify Antioxidant Activity In Vivo" Antioxidants 8, no. 5: 142. https://doi.org/10.3390/antiox8050142