Phytochemical Characterization, Antioxidant and In Vitro Cytotoxic Activity Evaluation of Juniperus oxycedrus Subsp. oxycedrus Needles and Berries
Abstract
:1. Introduction
2. Results and Discussion
2.1. Identification and Quantification of Phenolic Compounds
2.2. Mineral Composition of Joo Needles and Berries
2.3. Total Phenolic and Flavonoid Contents
2.4. Antioxidant Activity
2.5. Cytotoxicity of Methanolic Extracts of Joo Needles and Berries Against MDA-MB-468 and MCF7 Cell Lines
2.6. Clonogenic Assay of MDA-MB-468 and MCF-7 Treated with Methanolic Extracts of Joo Needles and Berries
2.7. Effect of Methanolic Extracts of Joo Needles and Berries on Antioxidant Enzyme Activities in MDA-MB-468 and MCF-7 Tumors Cell Lines
3. Materials and Methods
3.1. Chemicals and Reagents
3.2. Plant Material and Extracts Preparation
3.3. Total Phenolic Content
3.4. Total Flavonoid Content
3.5. HPLC-DAD Analysis
3.6. Mineral Content by ICP-AES
3.7. DPPH Radical Scavenging Assay
3.8. Radical Scavenging Activity Against the Radical ABTS+
3.9. Metal Chelating Activity
3.10. Reducing Power Assay (FRAP)
3.11. Cell Culture
3.12. MTT Assay
3.13. Cytotoxic Effect against Peripheral Blood Mononuclear Cells (PBMCs)
3.14. Clonogenic Formation Assay
3.15. Hydrogen Peroxide (H2O2) Content Determination
3.16. Malondialdehyde (MDA) Content Determination
3.17. Enzyme Activity Assays
3.17.1. Preparation of Cell Extracts for Antioxidant Enzyme Assays
3.17.2. Antioxidant Enzyme Assays
3.17.3. Protein Content Determination
3.18. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are available from the authors. |
Phenolic Compounds mg/100 g dw | Joo Needles | Joo Berries | ||
---|---|---|---|---|
Aqueous | Methanolic | Aqueous | Methanolic | |
Hydroxycinnamic acids | ||||
Caffeic acid | 8.68 ± 0.001 | 5.0 ± 0.001 | 11.4 ± 0.004 | 8.1 ± 003 |
p-Coumaric acid | 17.5 ± 0.001 | 21.2 ± 0.001 | ND | ND |
Hydroxybenzoic acids | ||||
Gallic acid | ND | ND | ND | ND |
Salicylic acid | 3398.1 ± 0.026 | 2942.7 ± 0.017 | 128.0 ± 0.010 | ND |
Syringic acid | ND | ND | ND | ND |
p-Hydroxybenzoic acid | 147.7 ± 0.009 | 416.6 ± 0.006 | 5.9 ± 0.003w | 2.9 ± 0.005 |
Flavonoids | ||||
Hesperidin | 278.8 ± 0.010 | 257.2 ± 0.006 | 147.0 ± 0.013 | 164.6 ± 0.009 |
Naringenin | 64.5 ± 0.004 | 18.4 ± 0.003 | 13.8 ± 0.007 | 19.0 ± 0.001 |
Rutin | 1080.5 ± 0.025 | 160.6 ± 0.007 | 8.1 ± 0.015 | 60.0 ± 0.011 |
Terpenes | ||||
Limonene | ND | ND | ND | ND |
Thymoquinone | ND | 81.4 ± 0.007 | 4.3 ± 0.009 | 37.5 ± 0.006 |
Mineral Content mg/kg dw | Joo Needles | Joo Berries |
---|---|---|
Macroelements | ||
Ca | 20,188 ± 1046 | 4609 ± 164 |
K | 7953 ± 188 | 2778 ± 89 |
Mg | 4539 ± 250 | 3413 ± 109 |
Na | 2312 ± 133 | 2415 ± 219 |
P | 1681 ± 170 | 1613 ± 43 |
Microelements | ||
Co | 0.35 ± 0.04 | 0.37 ± 0.04 |
Fe | 285.7 ± 49.07 | 59.66 ± 2.55 |
Mn | 79.91 ± 15.97 | 29.78 ± 0.57 |
Zn | 97.47 ± 16.16 | 59.01 ± 2.59 |
Cr | 0.60 ± 0.17 | ND |
Cu | 1.94 ± 0.43 | 1.91 ± 0.03 |
Se | 0.99 ± 0.36 | 1.28 ± 0.42 |
Heavy metals | ||
Cd | ND | ND |
Plant Material | Extraction Solvent | Polyphenols (mg GAE/g dw) | Flavonoids (mg QE/g dw) | Antioxidant Properties (IC50 Values; mg/mL) | Reducing Power (mg AAE/g dw) | ||
---|---|---|---|---|---|---|---|
DPPH | ABTS | Metal Chelating Activity | |||||
Joo N | Water | 147.29 ± 6.76 a | 28.66 ± 0.97 a | 0.12 ± 0.01 a | 0.47 ± 0.04 a | 1.53 ± 0.20 a | 89.70 ± 4.58 a |
Methanol | 292.52 ± 11.68 b | 54.58 ± 2.98 b | 0.05 ± 0.00 b | 0.12 ± 0.00 b | 1.59 ± 0.11 a | 139.14 ± 2.77 b | |
Joo B | Water | 28.11 ± 3.11 c | 3.20 ± 0.79 c | 0.96 ± 0.07 c | 1.61 ± 0.26 c | 0.96 ± 0.03 b | 257.99 ± 2.16 c |
Methanol | 131.48 ± 4.58 d | 8.28 ± 0.74 d | 0.09 ± 0.00 ab | 0.30 ± 0.01 ab | 1.89 ± 0.20 c | 941.81 ± 43.21 d |
Sample Tested | IC50 of Cytotoxic Activity Against Tumor Cells | % of Viability in PBMCs | ||||
---|---|---|---|---|---|---|
MDA-MB-468 | MCF-7 | PBMCs | Concentration (µg/mL) | |||
12.5 | 3.125 | 0.78125 | ||||
Joo N | 14.30 ± 3.3 a | 10.10 ± 1.40 a | >50 a | 91.34 ± 7.72 a | 110.40 ± 9.51 a | 126.82 ± 10.70 a |
Joo B | 6.40 ± 1.1 b | 5.20 ± 2.00 b | 49 a | 60.40 ± 7.68 a | 106.30 ± 1.89 a | 118.59 ± 5.56 a |
CisP | 0.20 ± 0.0 c | 2.20 ± 0.40 c | 0.27 b | 16.08 ± 3.39 b | 30.08 ± 3.58 b | 37.96 ± 3.44 b |
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Ben Mrid, R.; Bouchmaa, N.; Bouargalne, Y.; Ramdan, B.; Karrouchi, K.; Kabach, I.; El Karbane, M.; Idir, A.; Zyad, A.; Nhiri, M. Phytochemical Characterization, Antioxidant and In Vitro Cytotoxic Activity Evaluation of Juniperus oxycedrus Subsp. oxycedrus Needles and Berries. Molecules 2019, 24, 502. https://doi.org/10.3390/molecules24030502
Ben Mrid R, Bouchmaa N, Bouargalne Y, Ramdan B, Karrouchi K, Kabach I, El Karbane M, Idir A, Zyad A, Nhiri M. Phytochemical Characterization, Antioxidant and In Vitro Cytotoxic Activity Evaluation of Juniperus oxycedrus Subsp. oxycedrus Needles and Berries. Molecules. 2019; 24(3):502. https://doi.org/10.3390/molecules24030502
Chicago/Turabian StyleBen Mrid, Reda, Najat Bouchmaa, Youssef Bouargalne, Btissam Ramdan, Khalid Karrouchi, Imad Kabach, Miloud El Karbane, Abderrazak Idir, Abdelmajid Zyad, and Mohamed Nhiri. 2019. "Phytochemical Characterization, Antioxidant and In Vitro Cytotoxic Activity Evaluation of Juniperus oxycedrus Subsp. oxycedrus Needles and Berries" Molecules 24, no. 3: 502. https://doi.org/10.3390/molecules24030502