TOTAL PHENOLS CONTENT, FLAVONOID CONCENTRATION AND ANTIOXIDANT ACTIVITIES OF LEAVES EXTRACTS OF VITEX AGNUS-CASTUS L. GROWING WILD IN MOROCCOHTML Full Text
TOTAL PHENOLS CONTENT, FLAVONOID CONCENTRATION AND ANTIOXIDANT ACTIVITIES OF LEAVES EXTRACTS OF VITEX AGNUS-CASTUS L. GROWING WILD IN MOROCCO
Fatima EL kamari *, Amal Taroq, Yassine EL atki, Imane Aouam, Badiaa Lyoussi and Abdelfattah Abdellaoui
Laboratory of Physiology Pharmacology and Environmental Health, Department of Biology, Faculty of Sciences Dhar EL Mehraz, University Sidi Mohamed Ben Abdellah, B.P. 1796, Atlas, Fez, Morocco.
ABSTRACT: Vitex agnus-castus L. (Verbenaceae) commonly known as Chaste tree, is a shrub widely distributed in the Middle East and Southern Europe. Traditionally used for the treatment of several health problems and symptoms, such as premenstrual ones and spasmodic dysmenorrhea, certain menopausal conditions, insufficient lactation, and acne. Several reports have indicated that Vitex agnus-castus contained flavonoids, diterpenoids, and essential oils. To our knowledge, no data are available on the antioxidant activities of Moroccan Vitex agnus-castus. This work aims to evaluate the antioxidant activity of four extracts from leaves of Vitex agnus-castus growing wild in Morocco. Phytochemical tests of the extracts (ethanol, methanol, ethyl acetate, and water) were carried out qualitatively for the presence of Alkaloids, Tannins, Anthraquinones, Terpenes, and saponins. In-vitro antioxidant activity, total phenols content and total flavonoid content of different extracts were determined using spectrophotometric methods. The total phenolic content ranged from (10 ± 0.2 to 53.33 ± 1.38 mgEq GAE/g DW), and the total flavonoid concentrations varied from (13.66 ± 0.33 to 95.33 mg RE/gDW). Ethanolic extract of Vitex agnus-castus leaves has shown the highest phenols and flavonoid concentrations and strong antioxidant activity. A very strong positive correlation between the total antioxidant activity of the extracts and their content of phenols and flavonoids (P<0.05) is observed. Therefore, Moroccan Vitex agnus-castus L. can be regarded as promising candidates for natural plant sources of antioxidants with high value.
Antioxidant activity, Flavonoids, Vitex agnus-castus L., Phenols
INTRODUCTION: Free radicals and reactive oxygen species (ROS) are considered to be harmful to human health and play an essential causative role in disease initiation such as neurodegenerative disease, and cancer 1.
Thus, there is a growing interest in finding natural substances exhibiting antioxidant properties to substitute the synthetic ones, which were restricted due to their side effects. Several previous studies have reported that medicinal plants contain a large variety of free radical scavenging molecules such as phenols, anthocyanins, tannins, alkaloid and saponins which can play a significant role in the prevention and protection against many diseases 2.
Vitex agnus-castus L. (VAC) commonly known as Chaste tree 3 or Chasteberry 4 , is a small tree from Verbenaceae family, native to the Mediterranean and Western Asia. It is now cultivated all over the world, including the Southern part of the United States 5. Traditionally, this plant, particularly its fruit extract, has been used in the treatment of premenstrual syndrome (PMS), abnormal menstrual cycles, amenorrhea, mastodynia, hyperprolactinemia, premenstrual dysphoric disorder, lactation difficulties, and low fertility 6. The aromatic leaves are used as a spice 7. Previous studies on the phytochemical analysis of V. agnus-castus L. revealed the presence of essential oils 8, flavonoids, iridoids, and diterpenoids 9. It was reported that VAC was also used as a diuretic, digestive, anxiolytic and antifungal agent 10. Besides, different extracts of Vitex agnus-castus L. fruits have shown a significant cytotoxic, DNA damaging and apoptotic effects in MCF-7 human breast cancer cells 11. A recent study has demonstrated that the essential oils of fruits and leaves of Vitex agnus-castus have strong antibacterial activity against some bacteria causing nosocomial infections in the neonatal and intensive care rooms at the university hospital center of Fez, Morocco 12. This work aims to evaluate, for the first time, the antioxidant activity, flavonoids and total phenolic content of extracts of Vitex agnus-castus leaves from the Moroccan Middle Atlas.
MATERIALS AND METHODS:
Plants Materials: Plant material (leaves of Vitex agnus-castus) was collected during June October in 2016 (flowering period) in Khenifra. Identification was confirmed by Professor Amina Bari, botanist (Department of Biological Sciences, Faculty of Science, Sidi Mohammed Ben Abdellah University, Fes, Morocco). The material was dried for 7 to 10 days in the shade temperature and then ground.
Extract preparation: Ethanol (Etoh. E.), methanol (Metoh. E.), water (water E.) and ethyl acetate (Ethy Ac. E.) extractions were performed at the ratio of 10% (w/v) for 48 h under agitation for plant powder. Then the mixture was filtered through a filter paper (Whatman no. 1) and concentrated in vacuo at 45 °C, then stored at 4 °C for further use.
Phytochemical Analysis: Phytochemical tests of the extracts were carried out qualitatively for the presence of alkaloids, saponins, tannins, flavonoids anthraquinones, terpenes, according to the methods described by Edeoga et al., with slight modifications 13.
Total Antioxidant Capacity: The total antioxidant capacity of the extract was evaluated by the phosphomolybdenum method as described by Pavithra et al., with minor modifications 14. Briefly, a volume of 25 µL extract was added to 1 mL of reagent solution (0.6 mol/L sulphuric acid, 28 mmol/L sodium phosphate and 4 mmol/L ammonium molybdate). The mixtures were incubated at 95 °C for 90 min and then cooled to room temperature. The absorbance was measured at 695 nm. The total antioxidant activity was expressed as the number of equivalence of ascorbic acid (mg AAE/g DW).
Determination of Total Phenols Content (TPC): Total phenol content of the extract was determined by the Folin-Ciocalteu method described by Jadouali et al., with slight modifications 15. The 0.5 ml of a known dilution of the extract and 2 ml of 7% sodium carbonate solution were added to 2.5 ml of 10% (v/v) Folin - Ciocalteau reagent. The absorbance was read at 760 nm (Jasco v-530) after 2 h of reaction at room temperature in the dark. Gallic acid was used as a standard for the construction of calibration curve Fig. 1. Total phenols content was expressed as milligrams of gallic acid equivalents per gram dry weight (mg GAE/g DW).
FIG. 1: GALLIC ACID STANDARD CUVE FOR THE CALCULATION OF TOTAL PHENOLS CONTENT
Determination of Total Flavonoid Content: Total flavonoid content of extracts was measured by the aluminum chloride colorimetric assay 16. 1 ml of sample or rutin standard solution was added to a 10 mL volumetric flask containing 4 ml of distilled water. To the flask 0.30 ml 5% NaNO2 was added, after five minutes 0.3 ml 10% AlCl3 was added to react for 6 min. After that, 2 ml IM NaOH was added and the total was made up to 10 ml with distilled water. The solution was mixed, and absorbance was measured against the blank at 510 nm (Jasco v-530). Rutin was used as a standard for the construction of calibration curve Fig. 2. Total flavonoid content was expressed as mg Rutin equivalents per gram dry weight of each extract (mg RE/g DW).
FIG. 2: RUTIN STANDARD CUVE FOR THE CALCULATION OF TOTAL FLAVONOIDS CONTENT
DPPH Radical Scavenging Assay: The radical scavenging activity of the VAC extracts against 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical was measured using the method of Adebiyi et al., with minor modifications 17. 0.1 ml of various concentrations of the extracts or standard was added to 1.5 ml of an ethanolic solution containing 0.1 mmol of DPPH (2, 2-diphenyl-1picryl-hydroxyl). The absorbance of the mixture was measured at 517 nm with a spectrophotometer (Jasco V-530) after 30 min of incubation time at room temperature in the dark. The percentage of inhibition was calculated by the following equation:
% Inhibition = [(AC –AS)/AC] × 100
Where AC is the absorbance of the control, and AS is the absorbance of the sample. BHT served as positive control. The IC50 values were calculated as the concentration of extract providing 50% inhibition of DPPH radical.
Reducing Power Assay: The reducing capacity of the tested extracts was determined by the procedure of Oyaizu 18. 200 µl of the extract was mixed with 500µl of phosphate buffer (0.2M, pH 6.6) and 500 µl of potassium ferricyanide [K3Fe(CN)6] 1%. The obtained solution was incubated at 50 °C for 20 min.
The mixture was acidified with 500 µl of Trichloroacetic (TCA) 10% which was then centrifuged at 3000 rpm for 10 min. The upper layer of the solution (2.5 ml) was mixed with 500 µl of distilled water and 100 µl of FeCl3 (0.1%), and the absorbance was measured at 700 nm (Jasco v-530). Quercetin was used as a standard. The results were expressed as IC50 (mg/ml). IC50 (concentration corresponding 0.5 of absorbance) was calculated by plotting absorbance against the corresponding concentration.
Statistical Analysis: All tests were performed in triplicate and results were expressed as mean ± SD. The results were compared by one-way ANOVA followed by Tuckey-test, using the GraphPad Prism 5 (Microsoft Software). Differences at P<0.05 were considered to be significant. Pearson correlation was performed using the same test.
RESULTS AND DISCUSSION:
Phytochemical Analysis: Results of phytochemical screening of all the extracts are shown in Table 1. Flavonoids and phenols are present in all extracts, while saponins were present in water and ethanol extracts and absent in ethyl acetate and methanol extract. Alkaloids and terpenes were not detected in water and ethanol extract. Anthraquinones were observed only in the ethanol extract. These bioactive components have been shown to possess interesting biological activities such as antioxidant, antidiabetic, antibacterial, anti-inflammatory and many others 19, 20, 21.
TABLE 1: PHYTOCHEMICAL SCREENING TEST RESULTS
+ (present); - (Absent)
Extraction Yield and Total Antioxidant Capacity: The yields of extracts obtained from Vitex agnus-castus leaves using various solvents are shown in Table 2. Water gives the higher extraction yield with 24%, followed by methanol (22.44%), ethanol (10.02%) and ethyl acetate (6.12%). Our results are in agreement with those obtained by saglam et al., who reported that water gives the highest yield extract (38.06%) when compared with ethanol and n-hexane extracts of vitex agnus-castus leaves and fruits 22. This observation is also supported by results obtained from the extraction from Limnophila aromatica where the polar solvents, methanol, and water, gave the best extraction yields 23. It has been reported that the efficiency of the extraction depends on many parameters, including the extraction time and temperature, the volume and type of the solvents used 24, 25. The total antioxidant capacity assay showed that the ethanol extract had the most important activity with value 357.66 ± 5.08 mg equivalent to ascorbic acid/g DW. The water extract had a weak antioxidant activity.
TABLE 2: EXTRACTION YIELD AND TOTAL ANTIOXIDANT CAPACITY OF DIFFERENT EXTRACTS OF VITEX AGNUS-CASTUS LEAVES
(mg Eq A.As/gDW)
|Ethanol||10.02||357.66 ± 5.08a|
|Methanol||22.44||194 ± 1.41b|
|Ethyl acetate||6.12||138 ± 2.04c|
|Water||24||33.6 ± 1.16d|
Data are expressed as mean ± SEM of tree measurements. Different letters symbolized significant differences (P<0.05) by mean of the non-parametric Turkey-test.
Determination of Total Phenols Content: Most antioxidant activities from plant sources are derived from phenolic compounds 26. As we can see from Fig. 3, the amount of TPC of VAC extracts, measured by Folin-Ciocalteu method varied significantly (P<0.05) from 10 ± 0.2 to 53.33 ± 1.38 mg GAE/g DW. Pure ethanol produced extracts with the highest levels of total phenols content. Our finding is supported by Latoui et al., who demonstrated that ethanol behaved as a better solvent than methanol to extract from fruits of VAC the same classes of compounds whose concentrations in the extract reached after only 30 min maximum values (19.2 mg CAE/gDB TP and 5.4 mg CAE/Gdb OD) about 3-5-fold those obtained with methanol 27. Furthermore, several studies have shown that the amount of polyphenolics in plants depends on biological factors (genotype, organ, and ontogeny), as well as edaphic, and environmental (temperature, salinity, water stress and light intensity) conditions.
Besides, the solubility of phenolic compounds is governed by the type of solvent used, the degree of polymerization of phenolics, and their interaction28.
FIG. 3: PHENOLS CONTENT OF DIFFERENT EXTRACTS OF VITEX AGNUS-CASTUS LEAVES Different letters symbolized significant differences (P<0.05) by mean of the nonparametric Turkey-test
Determination of Total Flavonoid Content: The total flavonoid content was expressed as mg Rutin equivalents per gram dry weight of each extract (mg RE/g DW). Results can be seen from Fig. 4, the concentration of flavonoids in VAC extracts have ranged from 13.66 ± 0.33 to 95.33 ± 6.16 mg RE/g DW. The ethanol extract contains the highest content of flavonoids. The lowest flavonoids content is obtained in ethyl acetate and water extract. The concentration of flavonoids in plant extracts depends on the polarity of solvents used in the extract preparation 29. According to work performed by Latoui et al., the methanolic extract of VAC leaves from Tunisia contains 19.4 mg Catechin Equivalent/g dry biomass, which is below that found in the present study (48.66 ± 5.33 mg RE/g DW) 27. Our results have shown the richness of the VAC leaves for these bioactive molecules.
FIG. 4: FLAVONOIDS CONTENT OF DIFFERENT EXTRACTS OF VITEX AGNUS CASTUS LEAVES. Different letters symbolized significant differences (P<0.05) by mean of the nonparametric Turkey-test.
DPPH Radical Scavenging Activity: Antioxidant activity of V. agnus-castus extracts has been determined by two different test systems namely DPPH and reducing power (FRAP). Results are shown in Table 3, from which it can be observed that all the extracts possess radical scavenging potential. Ethanol has proved to be the most active of the extracts with (IC50 = 0.53 ± 0.006 mg/ml), while the water extract was the least (IC50 = 1.01 ± 0.019 mg/ml). None of the samples evaluated have shown activity as strong as the synthetic antioxidant BHT (0.10 ± 0.001 mg/ml).
An extract is considered to be active against free radicals if IC50 < 5 mg/ml 30. All our extracts have IC50 values of less than 5 mg/ml. Therefore, all the extracts for the solvents used are a possible good source of antioxidants. Also, we compared our results with an early report from Turkey, the antioxidant activity of methanolic extract of VAC fruits and leaves collected from Antalya was investigated by DPPH assay, methanol extracts of both leaves and fruits were found to possess strong antioxidant activity 31. Our data are supported by this result, the methanol extract of Moroccan leaves of VAC exhibit a good antioxidant activity with an IC50 value of 0.66 ± 0.012 mg/ml.
TABLE 3: ANTIOXIDANT ACTIVITY OF VITEX AGNUS CASTUS EXTRACTS, REDUCING CAPACITY (FRAP TEST) AND RADICAL SCAVENGING ACTIVITY (DPPH TEST) EXPRESSED IN IC50 (mg/ml)
|Ethanol||0.53 ± 0.006b||0.63 ± 0.004 c|
|Methanol||0.66 ± 0.012b||0.44 ± 0.011b|
|Ethyl Acetate||0.78 ± 0.020c||0.98 ± 0.016d|
|Water||1.01 ± 0.019d||2.02 ± 0.202e|
|BHT||0.10 ± 0.001a||0.12 ± 0.004a|
Data are the mean of three measurements ± SEM. In the column, Different letters symbolized significant difference (P<0.05) by mean of the nonparametric Turkey-test.
Reducing Power Activity: Regarding the reducing power assay, the strongest activity is exhibited by the methanol extract with (IC50 = 0.44 ± 0.011 mg/ml), followed by ethanol and ethyl acetate. However, all extracts are less effective than the synthetic antioxidant BHT.
Correlation of Antioxidant Activities with Flavonoids and Phenols Content: The correlation coefficients between flavonoids, phenols compounds and antioxidant activities of the four extract are shown in Table 3. Based on these results, it appears that there is a very strong positive correlation between the total antioxidant activity of the extracts and their content of phenols and flavonoids (P<0.05). Indeed, the high correlation between phenols content and antioxidant activity is well documented 32, 33. In previous studies, some phenolics were analyzed, and results demonstrated that especially casticin, caffeic acid and chlorogenic acid had been found in high amount in Vitex agnus-castus as antioxidant agents 34. Furthermore, Vitexin was determined in a significant amount in the leaves, as one of the essential antioxidant constituents of Vitex agnus-castus 31. In this study, we have noticed a low correlation in absolute value between phenols and flavonoids in plant extracts and reducing power activity. According to Yen et al., the reducing power is associated with some anthraquinones 35. However, just one of our extract contains anthraquinones.
TABLE 4: PEARSON CORRELATION COEFFICIENTS BETWEEN COMPOUNDS AND ANTIOXIDANT ACTIVITIES
CONCLUSION: In the current work, the total phenols content, total flavonoids content, and antioxidant activity have been investigated in the first time for Moroccan Vitex agnus-castus. The highest total phenolic and flavonoid contents are obtained from the ethanol extract of VAC leaves. Ethanol and methanol extracts have exhibited an interesting antioxidant activity. Based on these results, it could be concluded that Vitex agnus-castus is a new and inexpensive natural source of antioxidant substances which can combat oxidant damage and prevent pathogenesis of many diseases.
ACKNOWLEDGEMENT: The authors are profoundly thankful to the Department of Biology, of University Sidi Mohamed Ben Abdellah for providing all the necessary facilities to carry out this work.
CONFLICT OF INTEREST: The authors declare that there is no conflict of interest regarding the publication of this paper.
- Poprac P, Jomova K, Simunkova M, Kollar V, Rhodes CJ and Valko M: Targeting free radicals in oxidative stress-related human diseases. Trends in Pharmacological Sciences 2017; 38(7): 592-607.
- Sagbo IJ, Afolayan AJ and Bradley G: Antioxidant, antibacterial and phytochemical properties of two medicinal plants against the wound-infecting Asian Pacific Journal of Tropical Biomedicine 2017; 7(9): 817-825.
- Yushchyshena O and Tsurkan O: Phenolic compounds content in Vitex agnus-castus and V. cannabifolia Sieb. growing in Ukraine. Journal of Medicinal Plants Studies 2014; 2(5): 36-40.
- Šošić-Jurjević B, Ajdžanović V, Filipović B, Trifunovic S, Jaric I, Ristic N and Milosevi´c V: Functional morphology of pituitary-thyroid and -adrenocortical axes in middle-aged male rats treated with Vitex agnus castus essential oil. Acta Histochemica 2016; 118: 736-745.
- Habbab A, Sekkoum K, Belboukhari N, Cheriti A and Aboul-Enein HY: Essential oil chemical composition of Vitex agnus-castus from Southern-West Algeria and its antimicrobial activity. Current Bioactive Compounds 2016; 12: 51-60.
- Ahangarpour A, Najimi SA and Farbood Y: Effects of Vitex agnus-castus fruit on sex hormones and antioxidant indices in a D-galactose-induced aging female mouse model. Journal of the Chinese Medical Association 2016; 79: 589-96.
- Miguel M, Bouchmaaa N, Aazza S, Gaamoussi F and Lyoussi B: Antioxidant, anti-inflammatory and anti-acetylcholinesterase activities of eleven extracts of Moroccan plants. Fresenius Environmental Bulletin 2014; 23(6): 1-14.
- Neves RCS and Da Camara CAG: Chemical composition and acaricidal activity of the essential oils from Vitex agnus-castus (Verbenaceae) and selected monoterpenes. Anais da Academia Brasileira de Ciencias 2016; 88(3): 1221-1233.
- Allahtavakoli M, Honari N, Pourabolli I, Arababadi MK, Ghafarian H, Roohbakhsh A, Nadimi AE and Shamsizadeh A: Vitex agnus-castus extract improves learning and memory and increases the transcription of estrogen receptor α in the hippocampus of ovariectomized rats. Basic and Clinical Neuroscience 2015; 6(3): 185-192.
- Katiraee F, Mahmoudi R, Tahapour K , Hamidian G and Emami SJ: Biological properties of Vitex agnus-castus essential oil (Phytochemical component, antioxidant and antifungal activity). Biotech Health Sci 2015; 2(2): e26797.
- Aslantürk SÖ and Çelik AT: Antioxidant activity and anticancer effect of Vitex agnus-castus (Verbenaceae) seed extracts on MCF-7 breast cancer cells. Caryologia 2013; 66(3): 257-267.
- El-Kamari F, Taroq A, El- atki Y, Aouam I, Lyoussi B and Abdellaoui A: Chemical composition of essential oils from Vitex agnus-castus growing in Morocco and its in-vitro antibacterial activity against clinical bacteria responsible for nosocomial infections. Asian Journal of Pharmaceutical and Clinical Research 2018; 11(10): 365-368.
- Edeoga HO, Okwu DE and Mbaebie BO: Phytochemical constituents of some Nigerian medicinal plants. African Journal of Biotechnology 2005; 4(7); 685-688.
- Pavithra S and Banu N: Free radical scavenging activity and total antioxidant capacity of tin chlorophyllin from Morinda citrifolia Research Journal of Pharmacy and Technology 2017; 10(2): 453.
- Jadouali SM, Atifi H, Bouzoubaa Z, Majourhat K, Gharby S, Achemchem F, Elmoslih A, Laknifli A and Mamouni R: Chemical characterization, antioxidant and antibacterial activity of Moroccan Crocus sativus petals and leaves. Journal of Materials and Environmental Science 2018; 9(1): 113-118.
- Rahman MM, Islam MB, Biswas M and Alam AHM: In-vitro antioxidant and free radical scavenging activity of different parts of Tabebuia pallida growing in Bangladesh. BMC Research Notes 2015; 8: 621.
- Adebiyi OE, Olayemi FO, Ning-Hua T and Guang-Zhi Z: In-vitro antioxidant activity, total phenolic and flavonoid contents of ethanol extract of stem and leaf of Grewia carpinifolia. Beni-Suef University Journal of Basic and Applied Sciences 2017; 6(1): 10-14.
- Oyaizu M: Studies on products of browning reactions : Antioxidative activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 1986; 44: 307-315.
- Teugwa CM, Mejiato PC, Zofou D, Tchinda BT and Boyom FF: Antioxidant and antidiabetic profiles of two African medicinal plants: Picralima nitida (Apocynaceae) and Sonchus oleraceus (Asteraceae). BMC Comple-mentary and Alternative Medicine 2013; 13(1): 175.
- Kapewangolo P, Hussein AA and Meyer D: Inhibition of HIV-1 enzymes, antioxidant and anti-inflammatory activities of Plectranthus barbatus. Journal of Ethnopharmacology 2013; 149(1): 184-90.
- Pandey S: Preliminary phytochemical screening and in-vitro antibacterial activity of Bauhinia variegata against human pathogens. Asian Pacific Journal of Tropical Disease 2015; 5(2): 123-129.
- Sağlam H, Pabuçcuoğlu A and Kıvçak B: Antioxidant activity of Vitex agnus-castus extracts. Phytotherapy Research 2007; 21(11): 1059-1060.
- Do QD, Angkawijaya AE, Tran-Nguyen PL, Huynh LH, Soetaredjo FE, Ismadji S, and Ju YH: Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica. Journal of Food and Drug Analysis 2014; 22(3): 296-302.
- Chew KK, Khoo MZ, Ng SY, Thoo YY, Wan Aida WM and Ho CW: Effect of ethanol concentration, extraction time and extraction temperature on the recovery of phenolic compounds and antioxidant capacity of Orthosiphon stamineus International Food Research Journal 2011; 18(4): 1427-1435.
- Chigayo K, Mojapelo PEL, Mnyakeni-Moleele S and Misihairabgwi JM: Phytochemical and antioxidant properties of different solvent extracts of Kirkia wilmsii Asian Pacific Journal of Tropical Biomedicine 2016; 6(12): 1037-1043.
- Mahmoudi S, Khali M, Benkhaled A, Benamirouche K and Baiti I: Phenolic and flavonoid contents, antioxidant and antimicrobial activities of leaf extracts from ten Algerian Ficus carica varieties. Asian Pacific Journal of Tropical Biomedicine 2016; 6(3): 239-245.
- Latoui M, Aliakbarian B, Casazza AA, Seffen M, Converti A and Perego P: Extraction of phenolic compounds from Vitex agnus-castus Food and Bioproducts Processing 2012; 90: 748-754.
- Ksouri R, Megdiche W, Falleh H, Trabelsi N, Boulaaba M, Smaoui A and Abdelly C: Influence of biological, environmental and technical factors on phenolic content and antioxidant activities of Tunisian halophytes. Comptes Rendus Biologies 2008; 331(11): 865-873.
- Gao M and Liu CZ: Comparison of techniques for the extraction of flavonoids from cultured cells of Saussurea medusa World Journal of Microbiology and Biotechnology 2005; 21(8-9): 1461-1463.
- Abdillah S, Tambunan RM, Farida Y, Sandhiutami NMD and Dewi RM: Phytochemical screening and antimalarial activity of some plants traditionally used in Indonesia. Asian Pacific Journal of Tropical Disease 2015; 5(6): 454-57.
- Gökbulut A, Özhan O, Karacaoğlu M and Şarer E: Radical scavenging activity and vitexin content of Vitex agnus-castus leaves and fruits. Fabad Journal of Pharmaceutical Sciences 2010; 35: 85-91.
- Waheed I, Ahmad M, Syed NH and Ashraf R: Investigation of phytochemical and antioxidant properties of methanol extract and fractions of Ballota limbata (Lamiaceae). Indian Journal of Pharmaceutical Sciences 2014; 76(3): 251-256.
- Złotek U, Mikulska S, Nagajek M and Świeca M: The effect of different solvents and number of extraction steps on the polyphenol content and antioxidant capacity of basil leaves (Ocimum basilicum) extracts. Saudi Journal of Biological Sciences 2016; 23: 628-633.
- Hoberg E, Meier B and Sticher: Quantitative high performance liquid chromatographic analysis of casticin in the fruits of Vitex agnus-castus. Pharmaceutical biology 2001; 39(1): 57-61.
- Yen GC and Chuang DY: Antioxidant properties of water extracts from Cassia tora in relation to the degree of roasting. Journal of Agricultural and Food Chemistry 2000; 48(7): 2760-2765.
How to cite this article:
EL kamari F, Taroq A, EL atki Y, Aouam I, Lyoussi B and Abdellaoui A: Total phenols content, flavonoid concentration and antioxidant activities of leaves extracts of Vitex agnus-castus L. growing wild in Morocco. Int J Pharm Sci & Res 2019; 10(4): 1670-76. doi: 10.13040/IJPSR.0975-8232.10(4).1670-76.
All © 2013 are reserved by International Journal of Pharmaceutical Sciences and Research. This Journal licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.
F. E. Kamari *, A. Taroq, Y. E. Atki, I. Aouam, B. Lyoussi and A. Abdellaoui
Pharmacology and Environmental Health, Department of Biology, Faculty of Sciences Dhar EL Mehraz, University Sidi Mohamed Ben Abdellah, Atlas, Fez, Morocco.
15 July 2018
05 October 2018
08 October 2018
01 April 2019