Separation and Determination of Chemopreventive Phytochemicals of Flavonoids from Brassicaceae Plants
Abstract
:1. Introduction
2. Results
2.1. Extraction Yield
2.2. Hydrolysis of Extracts
2.3. Antioxidant Activity and Total Phenolics Contents in Plants Extracts
2.4. RP-ESI-UHPLC-MS/MS Method Validation
2.5. Determination of Phenolic Acids and Flavonoids in Brassicaceae Plants Extracts
3. Material and Methods
3.1. Analyzed Samples
3.2. Extract Preparation (Maceration)
3.3. Hydrolysis of Plant Extracts
3.4. Chemicals and Reagents for Preliminary Experiments
3.5. Apparatus
3.6. Antioxidant Activity
3.7. Determination of Total Phenolics Contents in Plants Extracts
3.8. TLC Analysis
3.9. RP-UHPLC-ESI-MS/MS Method to Determine Selected Flavonoids
3.10. Methods Validation
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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No | Sample Type | The Extraction Yield (EY) [%] ± SD a |
---|---|---|
1 | Radish–sprouts | 22.0 ± 2.11 |
2 | Radish–roots | 11.73 ± 1.18 |
3 | Radish–spots | 10.25 ± 1.04 |
4 | Radish–leaves | 6.22 ± 0.58 |
5 | Broccoli–sprouts | 19.96 ± 1.97 |
6 | Broccoli–flowers | 24.13 ± 2.42 |
7 | Brussels sprouts–leaves | 4.84 ± 0.35 |
8 | Kale–leaves | 16.86 ± 1.57 |
9 | Canola–roots | 8.00 ± 0.88 |
10 | Canola–spots | 11.88 ± 1.15 |
11 | Canola–leaves | 12.54 ± 1.19 |
12 | Conehead cabbage–leaves | 10.43 ± 1.02 |
13 | Kohlrabi–spots | 42.42 ± 4.15 |
14 | Kohlrabi–edible spot | 11.67 ± 1.09 |
15 | Kohlrabi–leaves | 14.23 ± 1.31 |
16 | Green cabbage–leaves | 16.06 ± 1.52 |
17 | Chinese cabbage–midvein | 23.22 ± 2.18 |
18 | Chinese cabbage–leaves | 13.59 ± 1.14 |
19 | Cauliflower–leaves | 7.27 ± 0.67 |
20 | Cauliflower–head | 18.73 ± 1.78 |
21 | Cauliflower–midvein | 28.75 ± 2.65 |
22 | Pak Choi cabbage–leaves | 17.49 ± 1.75 |
23 | Red cabbage–leaves | 15.10 ± 1.47 |
24 | Italian cabbage–leaves | 17.19 ± 1.57 |
25 | Kale–sprouts | 20.93 ± 2.13 |
Sample No | %RSA | TEAC [mM] | GAE [mg/mL] | Total Phenolic Content [mg/g] |
---|---|---|---|---|
1 | 86.73 ± 0.67 | 81.86 | 0.85 | 8.47 |
2 | 40.09 ± 0.18 | 24.44 | 0.47 | 4.67 |
3 | 41.12 ± 0.19 | 26.43 | 0.51 | 4.86 |
4 | 54.55 ± 0.64 | 14.36 | 0.59 | 5.87 |
5 | 84.35 ± 0.64 | 78.45 | 0.80 | 8.03 |
6 | 41.97 ± 0.09 | 18.22 | 0.44 | 4.37 |
7 | 40.29 ± 1.00 | 15.84 | 0.44 | 4.40 |
8 | 52.54 ± 0.18 | 23.14 | 0.66 | 6.62 |
9 | 30.89 ± 0.55 | 9.19 | 0.35 | 3.47 |
10 | 42.88 ± 0.28 | 13.39 | 0.41 | 4.10 |
11 | 51.24 ± 0.69 | 21.04 | 0.45 | 4.52 |
12 | 38.02 ± 2.20 | 47.77 | 0.48 | 4.77 |
13 | 45.97 ± 1.23 | 10.65 | 0.36 | 3.55 |
14 | 20.77 ± 1.83 | 23.53 | 0.40 | 3.99 |
15 | 63.89 ± 1.19 | 32.23 | 0.56 | 5.60 |
16 | 76.59 ± 0.09 | 87.16 | 0.63 | 6.27 |
17 | 80.23 ± 0.46 | 72.39 | 0.63 | 6.26 |
18 | 67.19 ± 0.18 | 45.87 | 0.47 | 4.67 |
19 | 45.86 ± 0.46 | 28.58 | 0.51 | 5.13 |
20 | 61.55 ± 0.28 | 43.98 | 0.43 | 4.33 |
21 | 42.43 ± 2.75 | 49.28 | 0.45 | 4.46 |
22 | 29.98 ± 0.94 | 27.35 | 0.43 | 4.33 |
23 | 58.12 ± 0.92 | 48.90 | 0.64 | 6.44 |
24 | 56.89 ± 0.64 | 71.25 | 0.59 | 5.89 |
25 | 60.65 ± 0.09 | 94.73 | 0.77 | 7.72 |
Analyte | Linear Range (ng/g) | R2 | LOD/LOQ (ng/g) | Nominal Concentration (ng/g) | Intra-Day | Inter-Day | Matrix Effect ME (%) | ||
---|---|---|---|---|---|---|---|---|---|
CV a (%) | RE b (%) | CV a (%) | RE b (%) | ||||||
3,4-DHBA | 0.4–500 | 0.9951 | 0.13/0.4 | 300 | 2.14 | 1.36 | 3.63 | 2.51 | −4.58 |
10 | 3.84 | 2.41 | 5.47 | 3.96 | −6.98 | ||||
α-HHA | 0.4–500 | 0.9924 | 0.13/0.4 | 300 | 4.12 | −2.15 | 6.18 | −3.74 | −8.47 |
10 | 5.47 | −3.47 | 7.56 | −5.25 | −7.36 | ||||
DOPAC | 0.4–500 | 0.9974 | 0.13/0.4 | 300 | 1.73 | −4.28 | 3.58 | −5.87 | 2.57 |
10 | 3.69 | −5.39 | 5.17 | −6.39 | 4.65 | ||||
4-HBA | 0.4–500 | 0.9936 | 0.13/0.4 | 300 | 2.58 | −2.76 | 3.81 | −4.85 | 5.36 |
10 | 4.69 | −3.33 | 4.83 | −6.97 | 6.17 | ||||
CA | 0.8–500 | 0.9947 | 0.26/0.8 | 300 | 1.93 | 2.15 | 4.51 | 4.28 | −5.12 |
10 | 2.17 | 4.87 | 6.37 | 5.78 | −6.47 | ||||
HA | 0.4–500 | 0.9991 | 0.13/0.4 | 300 | 4.83 | −1.79 | 7.42 | −3.69 | −2.14 |
10 | 5.39 | −3.65 | 8.69 | −4.83 | −6.07 | ||||
3-HBA | 0.4–500 | 0.9989 | 0.13/0.4 | 300 | 2.11 | −0.94 | 4.41 | −2.81 | 3.19 |
10 | 4.78 | −2.75 | 5.93 | −5.12 | 5.47 | ||||
3-HPA | 0.4–500 | 0.9978 | 0.13/0.4 | 300 | 4.32 | 3.54 | 6.71 | 4.32 | 2.58 |
10 | 6.71 | 4.39 | 8.54 | 6.96 | 4.96 | ||||
HVA | 0.8–500 | 0.9990 | 0.26/0.8 | 300 | 2.95 | −3.27 | 4.33 | −5.63 | −3.96 |
10 | 3.69 | −5.83 | 7.05 | −8.11 | −5.87 | ||||
3,4-HPPA | 20–500 | 0.9963 | 6.67/20 | 300 | 0.87 | −2.94 | 3.82 | −4.75 | −4.12 |
50 | 2.47 | −4.65 | 4.57 | −6.35 | −6.05 | ||||
p-COA | 0.4–500 | 0.9952 | 0.13/0.4 | 300 | 3.98 | 1.23 | 5.71 | 2.85 | 4.78 |
10 | 4.72 | 3.67 | 6.87 | 5.78 | 5.39 | ||||
FA | 0.4–500 | 0.9932 | 0.13/0.4 | 300 | 2.56 | 2.26 | 4.41 | 4.59 | −1.47 |
10 | 4.78 | 4.63 | 5.89 | 6.32 | −4.36 | ||||
ERC | 0.4–500 | 0.9981 | 0.13/0.4 | 300 | 3.66 | −3.15 | 4.87 | −4.59 | 3.97 |
10 | 5.21 | −3.98 | 6.39 | −5.93 | 5.82 | ||||
LIQ | 0.4–500 | 0.9994 | 0.13/0.4 | 300 | 4.17 | −4.71 | 5.71 | −6.47 | 2.74 |
10 | 6.41 | −5.02 | 8.53 | −7.89 | 4.69 | ||||
RUT | 0.4–500 | 0.9993 | 0.13/0.4 | 300 | 3.25 | 2.47 | 4.52 | 4.52 | 3.55 |
10 | 4.17 | 3.69 | 5.36 | 6.36 | 5.41 | ||||
TAX | 0.4–500 | 0.9947 | 0.13/0.4 | 300 | 2.87 | 4.52 | 4.23 | 5.29 | 2.69 |
10 | 4.63 | 5.87 | 6.87 | 7.85 | 3.74 | ||||
BA | 0.8–500 | 0.9968 | 0.26/0.8 | 300 | 1.85 | −2.78 | 2.54 | −3.89 | 4.11 |
10 | 3.64 | −5.69 | 4.85 | −7.25 | 5.07 | ||||
NRI | 0.8–500 | 0.9920 | 0.26/0.8 | 300 | 2.87 | −3.21 | 4.41 | −4.25 | −6.87 |
10 | 3.54 | −4.87 | 6.52 | −6.78 | −8.15 | ||||
NARG | 0.8–500 | 0.9994 | 0.26/0.8 | 300 | 0.47 | −5.70 | 1.23 | −6.23 | 4.25 |
10 | 3.69 | −6.42 | 4.32 | −8.52 | 5.78 | ||||
HSD | 0.4–500 | 0.9987 | 0.13/0.4 | 300 | 4.21 | 2.14 | 6.05 | 3.46 | 5.23 |
10 | 5.87 | 4.78 | 7.93 | 4.85 | 6.90 | ||||
NHSD | 0.4–500 | 0.9965 | 0.13/0.4 | 300 | 3.78 | 3.58 | 5.25 | 5.39 | 2.14 |
10 | 4.93 | 4.96 | 6.38 | 6.87 | 4.79 | ||||
FIS | 0.4–500 | 0.9971 | 0.13/0.4 | 300 | 2.41 | 1.64 | 4.21 | 2.58 | −3.67 |
10 | 5.87 | 2.83 | 7.28 | 4.65 | −4.52 | ||||
LQG | 0.4–500 | 0.9939 | 0.13/0.4 | 300 | 4.28 | −5.22 | 6.48 | −6.87 | −6.21 |
10 | 6.14 | −5.89 | 8.52 | −7.41 | −7.56 | ||||
ERI | 0.4–500 | 0.9914 | 0.13/0.4 | 300 | 5.02 | 1.74 | 6.35 | 2.69 | −5.47 |
10 | 5.89 | 3.69 | 6.98 | 4.78 | −8.73 | ||||
QUE | 0.8–500 | 0.9975 | 0.26/0.8 | 300 | 2.73 | 4.14 | 4.11 | 6.15 | −5.21 |
10 | 3.85 | 5.63 | 5.28 | 7.25 | −6.78 | ||||
NAR | 0.4–500 | 0.9993 | 0.13/0.4 | 300 | 1.73 | 2.54 | 2.55 | 4.32 | 1.63 |
10 | 4.62 | 4.36 | 5.87 | 5.69 | 2.87 | ||||
HST | 0.8–500 | 0.9945 | 0.26/0.8 | 300 | 3.98 | −5.32 | 3.69 | −6.14 | 3.69 |
10 | 5.87 | −6.12 | 7.21 | −8.25 | 5.74 | ||||
FOR | 0.4–500 | 0.9968 | 0.13/0.4 | 300 | 2.02 | −1.56 | 4.20 | −3.69 | 2.46 |
10 | 4.96 | −3.54 | 5.93 | −4.75 | 6.89 | ||||
PIN | 0.4–500 | 0.9972 | 0.13/0.4 | 300 | 1.52 | 2.47 | 2.47 | 3.62 | 1.25 |
10 | 3.69 | 3.69 | 4.89 | 5.55 | 4.98 | ||||
GLB | 0.4–500 | 0.9954 | 0.13/0.4 | 300 | 0.57 | 4.52 | 1.47 | 5.87 | −3.57 |
10 | 3.58 | 5.69 | 5.63 | 6.45 | −4.82 |
No | Sample Type | Latin Name |
---|---|---|
1 | Radish–sprouts | Raphanus sativus var. Sativus |
2 | Radish–roots | Raphanus sativus var. Sativus |
3 | Radish–spots | Raphanus sativus var. Sativus |
4 | Radish–leaves | Raphanus sativus var. Sativus |
5 | Broccoli–sprouts | Brassica oleracea L. var. italica Plenck |
6 | Broccoli–flowers | Brassica oleracea L. var. italica Plenck |
7 | Brussels sprouts–leaves | Brassica oleracea L. var. gemmifera (DC.) Zenker |
8 | Kale–leaves | Brassica oleracea L. var. sabellica L. |
9 | Canola–roots | Brassica napus L. |
10 | Canola–spots | Brassica napus L. |
11 | Canola–leaves | Brassica napus L. |
12 | Conehead cabbage–leaves | Brassica oleracea ‘Cour di Bue Grosso’ |
13 | Kohlrabi–spots | Brassica oleracea var. gongylodes L. |
14 | Kohlrabi–edible spot | Brassica oleracea var. gongylodes L. |
15 | Kohlrabi–leaves | Brassica oleracea var. gongylodes L. |
16 | Green cabbage–leaves | Brassica oleracea L. var. capitata L. |
17 | Chinese cabbage–midvein | Brassica rapa L. subsp. pekinensis |
18 | Chinese cabbage–leaves | Brassica rapa L. subsp. pekinensis |
19 | Cauliflower–leaves | Brassica oleracea L. var. botrytis L. |
20 | Cauliflower–head | Brassica oleracea L. var. botrytis L. |
21 | Cauliflower–midvein | Brassica oleracea L. var.botrytis L. |
22 | Pak Choi cabbage–leaves | Brassica rapa L. subsp. chinensis Hanelt |
23 | Red cabbage–leaves | Brassica oleracea var. capitata f. rubra |
24 | Italian cabbage–leaves | Brassica oleracea L. var. sabauda L. |
25 | Kale–sprouts | Brassica oleracea L. var. sabellica L. |
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Bajkacz, S.; Ligor, M.; Baranowska, I.; Buszewski, B. Separation and Determination of Chemopreventive Phytochemicals of Flavonoids from Brassicaceae Plants. Molecules 2021, 26, 4734. https://doi.org/10.3390/molecules26164734
Bajkacz S, Ligor M, Baranowska I, Buszewski B. Separation and Determination of Chemopreventive Phytochemicals of Flavonoids from Brassicaceae Plants. Molecules. 2021; 26(16):4734. https://doi.org/10.3390/molecules26164734
Chicago/Turabian StyleBajkacz, Sylwia, Magdalena Ligor, Irena Baranowska, and Bogusław Buszewski. 2021. "Separation and Determination of Chemopreventive Phytochemicals of Flavonoids from Brassicaceae Plants" Molecules 26, no. 16: 4734. https://doi.org/10.3390/molecules26164734