Variation in Phenolic Composition, Antioxidant Activity and Cyclic Voltammetry Study of Juncus rigidus Desf. (Juncaceae) Root Extracts according Flowering and Ripening Period

 

Mimouna Hani^1,3, Chérifa Boubekri^2,3*, Touhami Lanez^1,3

¹University of Echahid Hamma Lakhdar, Faculty of Exact sciences, PO Box 789, 39000, El Oued, Algeria.

²University Mohamed Khider, Faculty of Exact sciences, Natural and Life Sciences,
Department of Material sciences, BP145 RP Biskra 07000 Algeria.

³VTRS Laboratory, B.P.789, 39000, El Oued, Algeria.

*Corresponding Author E-mail: cherifa.boubekri@univ-biskra.dz

 

 

INTRODUCTION:

The Juncus genus is the most prevalent in the Juncaceae family, comprising over 300 species distributed across the continents, mostly found in salt marshes and in poorly drained soils. Several Juncus species are used for herbal medicine as sedatives and for treating various health issues, such as insomnia. This health benefit can be attributed to the presence of different classes of compounds including flavonoids, coumarins, sterols, phenolic acids, stilbenes and phenanthrens¹ which are commonly present in this genus².

 

Therefore, systematic studies have recently been carried out to identify plants with anti-radical potential for human consumption³. In recent years, numerous researchers have paid attention to bioactive substances, the most important of which are phenolic compounds that play a main role in protecting human health⁴. Phenolic compounds are secondary metabolites which are synthesized in plants⁵. It has potent antioxidant property, thereby which plays the important role in treatment of many of the diseases⁶.

 

Polyphenolic can interfere with the oxidation process by different mechanism⁷. As plants produce a lot of antioxidants to control the oxidative stress caused by sun beams and oxygen⁸. The oxidative stress is one of the most common pathogens currently time, it produces of imbalance between the formation and neutralization of reactive free radicals, this last are continuously neutralized and produced in our body to maintain the constant internal environment redox state⁹. Damage mediated by free radicals results in the disruption of membrane fluidity, protein denaturation, DNA mutation and lipid peroxidation and alteration of platelet functions¹⁰.

 

Antioxidant activity in food and beverages has become one of the most interesting features in the science community¹¹. The isolation and identification of antioxidant compounds in plant content is a long process, expensive and difficult. Therefore, researchers desire to develop methods that can provide rapid, reliable and direct measurement¹².

 

Several assays have been frequently used to estimate antioxidant capacities in fresh fruits and vegetables and their products¹³. This study aimed to compare the variation in the composition of the root of Juncus rigidus, collecting during two periods, by determining total phenolic content and total flavonoid content, and total flavonols contents and to investigate the antioxidant activity using different methods such as DPPH radical (1,1-diphenyl-2-picrylhydrazyl), FRAP (ferric reducing antioxidant power) and phosphomolybdenum assay. The electrochemical behavior was also studied using cyclic voltammetry method.

 

MATERIALS AND METHODS:

Chemical reagents:

All reagents were used for analysis, obtained from (Sigma; Aldrich) and (Merck Co.) : Ethanol, acetone, methanol, Folin-Ciocalteu reagent, sodium carbonate, gallic acid, aluminium trichloride, quercetin, sodium acetate, DPPH, ascorbic acid, sulphuric acid, sodium phosphate, ammonium molybdate, TPTZ (2,4,6-tripyridyl-s-triazine), iron (III) chloride hexahydrate (FeCl₃.6H₂O), hydrochloric acid.

 

Plant material:

Juncus rigidus plant was identified by the botanist Chehma (Laboratory for the protection of ecosystems in arid and semi-arid zones, Ouargla University). The harvest of the plant was carried out according to two periods (the period of flowering and the period of ripening) from the region of El-Oued. After the separation of the other aerial parts of the plant, roots were washed with tap water, drying in room temperature without exposure to sunlight or humidity, powdered using an electric grinder and conserved in boxes until uses.

 

Extract Preparation:

20 g of each dried powdered sample were successively mixed with 200 ml of each of the following solvents: ethanol and acetone for 24 h at room temperature and under magnetic stirring. After filtration, these four solutions were evaporated by rotary evaporator type Buchi R-200 at 30°C until obtaining a dry extract. All extracts were stored at 4 °C for further use. The yield of each extract was calculated.

 

Phytochemical analysis:

Total phenolic content (TPC):

Determination of total phenolic content in all samples was carried out according to the Folin-Ciocalteu assay reported by Li and al¹⁴ with simple modifications. Briefly, after the dilution of each dry extract in absolute ethanol to obtain the concentration of 1 mg/mL, 300 µL was mixed with 1.5 mL of Folin-Ciocalteu reagent diluted 10 times with distilled water, and the reaction mixture is left for 8 minutes at room temperature, so we add 1.2 mL of sodium carbonate Na₂CO₃ at a concentration of 7.5% to the previous mixture, which we allowed to stand at room temperature for 30 minutes. The absorbance was read using a UV-visible spectrophotometer at the wavelength of 765 nm. The calibration curve was reported using the gallic acid solution as a standard using the concentration range from (0.001-0.01) mg/mL. Total phenolic content was expressed as milligrams of gallic acid equivalents per gram of dry extract (mgGAE/g).  Analyses for each extract were repeated in triplicate.

 

Total flavonoid content (TFC):

The total flavonoid content in all root extracts of Juncus rigidus plant was quantified by aluminum trichloride method reported by Bahrun and al¹⁵. 1 mL of each extract (1 mg/mL) was added to 1 mL of AlCl₃ solution prepared previously in methanol at the concentration of 2%. The reaction mixture is allowed to react for 30 minutes at room temperature and the absorbance is read using a UV-visible spectrophotometer at a wavelength of 430 nm. The calibration curve was reported using the quercetin solution as a standard using the concentration range from (0.001-0.01) mg/mL. Total flavonoid content was expressed as milligrams of quercetin equivalent per gram of dry extract mgQE/g of dry extract. Analyses for each extract were repeated in triplicate.

 

Total flavonol content (TFLC):

Total flavonols in the dry extract of the plant were determined by the method reported by Formagio and al. with simple modification¹⁶. In this assay, 400 µL of extract solution was added to 400 µL of aluminum trichloride (2%) prepared in ethanol, and 0,6 mL of sodium acetate with a concentration of (50g/L) was also added. the mixture is left to react for 2,5 h at room temperature. and the absorbance was read using a UV-visible spectrophotometer at a wavelength of 440 nm. The calibration curve was reported using the quercetin solution as a standard using the concentration range from (0.01-0.1) mg/mL. Total flavonol content was expressed as milligrams of quercetin equivalent per gram of dry extract mgQE/g of dry extract. Analyses for each extract were repeated in triplicate.          

Antioxidant activity:

DPPH radical scavenging assay:

The hydrogen atom or electron donation ability of the extract was measured from the bleaching of purple-colored MeOH solution of DPPH. This spectrophotometric assay uses stable radical 1,1-Diphenyl-2-picrylhydrazyl (DPPH) as a reagent^17,18. To evaluate the free radical scavenging activity, we used the method cited by Que et al. with some modifications¹⁹. 1 mL of DPPH radical solution (4%) prepared in MeOH was added to 1 mL of extract solution prepared with different concentrations. The mixtures were incubated at room temperature for 30 min and the absorbance was read against a blank using a UV-visible spectrophotometer at a wavelength of 517 nm. The inhibition of free radical DPPH in percent was calculated according to the formula:

 

Where A control is the absorbance of the control and A sample is the absorbance of the test sample, extract concentration providing 50% inhibition (IC50) was calculated from the graph plotted inhibition percentage, tests were carried out in triplicate. Analyses for each extract were repeated in triplicate.

 

Total antioxidant activity assay (TAC):

The total antioxidant activity of the extract solution was determined by the method described by Amo-Mensah and al. with some modifications. Phosphomolybdenum (PPM) reagent was prepared by combining 0.6 M sulphuric acid, 28 mM sodium phosphate, and 4 mM ammonium molybdate²⁰, 0.1 mL of each extract was mixed with 1 mL of the reagent solution; the mixture was incubated in a water bath at 95°C for 90 min, after cooling to 25°C, the absorbance was read at 695 nm. Ascorbic acid was employed as standard. Analyses for each extract were repeated in triplicate.

 

Ferric reducing antioxidant power assay (FRAP):

The ferric reducing antioxidant power (FRAP) assay provides a measure of the reducing ability of the plant extracts²¹. Ferric reducing antioxidant power assay of all extracts was evaluated using the ferric-reducing ability described by Benzie and Strain²². A potential antioxidant will reduce the ferric ion (Fe³⁺) to the ferrous ion (Fe²⁺); the latter forms a blue complex (Fe²⁺/TPTZ), which increases the absorption at 593 nm. Briefly, the FRAP reagent was prepared by mixing acetate buffer (300 mM, pH 3.6), a solution of 10 mM TPTZ in 40 mM HCl, and 20 mM FeCl₃ at 10:1:1 (v/v/v)²³. 3 mL of FRAP reagent were added to 100 μL of extract solution, the mixture was allowed to stand 30 min at room temperature the absorbance was read at 593 nm. The standard calibration curve was prepared with various concentrations of FeSO₄. Analyses for each extract were repeated in triplicate.

Cyclic voltammetry assay:

Cyclic voltammetry is a unique technique for the electrochemical characterization of compounds by providing data about their oxidation/reduction potentials²⁴. In an electrochemical cell of 25 mL, we mixed 20 mL of phosphate buffer at pH=7 (0,2 M) as a supporting solution and 5 mL of extract solution. We placed a glassy carbon working electrode, a platinum wire counter electrode, and Hg/Hg₂Cl₂ reference electrode. The electrochemical cell was related to PGP 301 potentiostat. A cyclic voltammogram was obtained by one single cycle performed at a scan rate of 100 mV.S-1 and therefore the potential was swept in direct scanning mode ranging from 00 to +1000 mV. Ascorbic acid was used as a standard²⁵.

 

Statistical analysis:

Each analysis is performed in triplicate and results were reported as the mean and standard deviation (SD).

 

RESULTS AND DISCUSSION:

Extraction yield:

The production from chemical extraction depends on several different factors; the most important is the form of solvent with different polarities and chemical sample compositions²⁶. The yield (%) was calculated as follows:

 

 

The extraction of the roots of the Juncus rigidus plant during the ripening and flowering period was carried out under the same conditions of time and temperature. There is no previous research done on the effect of solvents on the roots of the species Juncus rigidus. The determination of a suitable solvent is necessary to optimize the recovery of phenolic and flavonoid compounds from the plant²⁷. The positive relationship between solvent and various quantities of crude extracts indicates that the roots of the plant have a different quantity of soluble plant chemicals which require a highly specific isolation solvent. Different biological compounds have different polarities and can be extracted by a polarity index solvent^28,29. This research was carried out in two periods; ripening and flowering periods. We used two polar solvents for the extraction which are ethanol and acetone to compare their effect on the extraction yield and the solubility of the chemical compounds present in the extracts. The results showed that the extraction solvents and growing period affected the extraction yield (Fig.1, Tab.1) ; the highest yield of extraction was obtained in ethanol extract (58,5%) compared to acetone extract (24,6%) during ripening stage.

 

 

 

Table 1. Yield of the extraction of the roots of Juncus rigidus extracts during flowering and ripening period.

 

The highest yield of extraction during the flowering period was also obtained in ethanol extract (47,4%), acetone extract was the lowest one, indicating that the biomolecules in the roots of J. rigidus are more soluble in ethanol. The extraction efficiency favors the highly polar solvents³⁰.

 

Figure 1. Yield of the extraction of the roots of Juncus rigidus extracts during flowering and ripening period

 

The yield of extraction was highest during the ripening period than the flowering period, which indicates that the composition of the plant changed during the growing stage³¹.

 

Total phenolic content (TPC):

the structures of polyphenols are very diverse and very varied. Thus, choosing of extraction solvent has a great potential effect on the recovery of phenolic compounds and the measured quantity of total phenolic content³². Total phenolic contents in plants are also affected with used part^33-34. Using the obtained standard curve equation of the gallic acid (y = 11.848 x - 0.274 with R² = 0.9989) we can calculate the total phenolic contents in all samples, expressed as mg GAE /g of dry extract. The results are shown in Table 2.

 

Table 2.  Total phenolic content of the roots of Juncus rigidus extracts during flowering and ripening period

 

The obtained results were significant and showed that the highest amount of TPC in the roots of J. rigidus was observed in acetone extract (87,249 ± 0.005 mg GAE/g) during the flowering period which decreased using ethanol (66,289 ± 0.001 mg GAE/g). Also, during the ripening period acetone extract had the highest amount of total phenolic contents (69,328 ± 0.015 mg GAE/g), and ethanol extract was the lowest (42,150 ± 0.003 mg GAE/g). The amount of TPC using ethanol as an extraction solvent was less than acetone (figure 2). Acetone has the lowest polarity than ethanol but recorded the highest value³⁵. The variations in phenolic content in the plant throughout the growing stages can be explained by biochemical changes occurring during growth as well as an immune response toward biotic and abiotic stresses³⁶.

 

Figure 2. Total phenolic contents of the roots of Juncus rigidus extracts during flowering and ripening period

 

Solvent polarity played a pivotal function in all extraction studies. The net molecular polarities of solvents were measured by their dipole moments. The separation of components depends on the polarity of both solvent and component³⁷.

 

Total flavonoid content (TFC):

Total favonoid contents of the roots of Juncus rigidus extracts of the studied samples were calculated using standard curve equation of the quercetin (y = 40,31x - 0,0242 with R² = 0.9952), expressed as mgQE/g of dry extract, the results are shown in table 3.

 

Table 3. Total flavonoid content of the roots of Juncus rigidus extracts during flowering and ripening period

 

It is very clear that all the results recorded were significant and indicate the presence of flavonoids in the roots of Juncus rigidus. By comparing the effect of the two polar solvents, it is noted that in both periods acetone gives the highest amount of total flavonoids content than ethanol (figure 3). The amount of total flavonoids was higher in flowering stage using acetone as extraction solvent (28,109 ± 0,041 mg QE/g) which was lowest in ethanol extract (20,415 ± 0,074 mg QE/g). This results decrease in ripening period, (23,775 ± 0,003 mg QE/g) in acetone extract and (16,193 ± 0,06 mg QE/g) in ethanol extract. Many researchers have observed the changes in amounts of total flavonoid contents of other plant tissues during development and period of growth^38,39

 

Figure 3. Total flavonoid contents of the roots of Juncus rigidus extracts during flowering and ripening period

 

Total flavonol content (TFlC):

Total flavonol contents of the roots of Juncus rigidus extracts of the studied samples were calculated using standard curve equation of the quercetin (y = 15,247x + 0,0972 with R² = 0,9993), expressed as mgQE/g of dry extract, the results are shown in table 4.

 

Table 4. Total flavonol content of the roots of Juncus rigidus extracts during flowering and ripening period

 

The flavonols are a subclass of the flavonoid family, they are widely diffused in higher plants, where they are uniformly distributed in fruits, flowers, leaves and stems⁴⁰. They are typical and promising phytochemicals belonging to the flavonoids. They have been recognized to exert various effects⁴¹. Figure 4 showed that a total amount of flavonols was higher in acetone extracts during flowering stage (33,195 ± 0,013 mg QE/g) and ripening stage (29,107 ± 0,002 mg QE/g) witch decrease in ethanol extract in the two periods.

 

Figure 4. Total flavonol contents of the roots of Juncus rigidus extracts during flowering and ripening period

 

These results are proportional to those recorded for the total phenolic contents and the total flavonoid contents. The highest concentration in TPC, TFC and TFLC was obtained using acetone as solvent of extraction and the best result was obtained during flowering stage. These results are in agreement with those found by many researchers, however the rate of total phenolics, total flavonoids and total flavonols reached is maximum during the flowering period. Seasonal variations in photoperiod, light intensity and temperature can significantly alter the levels of phenolic compounds in plants⁴².

 

Antioxidant activities:

Generally, antioxidant measurements can be related either to the capacity of extracts to directly transfer hydrogen to a radical or to donate electrons⁴³. Due to the complexity of some plant extracts, the use of several different methods is recommended for the evaluation of antioxidant activity⁴⁴.

 

DPPH radical scavenging activity:

The DPPH test is a method widely used in the analysis of antioxidant activity. Originally, DPPH• is a stable free radical. This stability is due to the delocalization of free electrons. The presence of these DPPH• radicals gives rise to a dark violet coloring of the solution⁴⁵. The DPPH radical scavenging activity of Juncus rigidus is affected by solvent polarity and harvest time. Table 5 showed that the most effective extract is the one with the lowest IC50 value. during ripening stage acetone extract of the roots of Juncus rigidus had the highest DPPH scavenging activity (0,014 ± 0,0003 mg/ml) than ethanol extract (0,028 ± 0,004 mg/ml) and also during flowering stage acetone extract had the highest DPPH scavenging activity (0,041 ± 0.002 mg/ml) than ethanol extract (0,078 ± 0,003 mg/ml).

 

Table 5. DPPH radical scavenging activities of the roots of Juncus rigidus extracts during flowering and ripening period

 

In comparing the two harvest periods, the DPPH scavenging activities of the roots of Juncus rigidus were significantly higher during the ripening period (figure 5).

 

Figure 5. DPPH scavenging avtivity of the roots of Juncus rigidus extracts during flowering and ripening period

 

Several research works have shown that the antioxidant activity of the extracts of the plants varies according to the harvest period^46-49 and according to the solvent polarities^50-52.

Ferric reducing antioxidant power (FRAP)

The harvest period and the polarity of the extraction solvents influenced the ferric-reducing antioxidant power of the roots of Juncus rigidus as shown in (table 6, figure 6). Antioxidants, which can effectively reduce prooxidants, can also effectively reduce Fe³⁺ to Fe²⁺. Therefore, the reducing power of a compound provides important information about its antioxidant activity⁵³.

 

Table 6. Ferric reducing antioxidant activities of the roots of Juncus rigidus extracts during flowering and ripening period

 

The results revealed that the acetone extracts of the roots of Juncus rigidus exhibited the highest FRAP value than ethanol extracts in both stages and the FRAP activity during the ripening period was higher than the activity obtained during the flowering period.

 

Figure 6. Ferric reducing antioxidant activities of the roots of Juncus rigidus extracts during flowering and ripening period

 

Total antioxidant capacity assay (TAC):

The basic principle to assess the antioxidant capacity through phosphomolybdenum assay includes the reduction of Mo (VI) to Mo (V) by the plant extract possessing antioxidant compounds^54,55. Table 7 showed that the highest total antioxidant capacity was exhibited by the acetone extract of roots of Juncus rigidus in both periods, while ethanol extract showed low total antioxidant activity. By comparing the two harvest periods, the total antioxidant capacity was strong during the ripening period (figure 7).

 

Table 7. PPM activities of the roots of Juncus rigidus extracts during flowering and ripening period

 

This result is in agreement with that found for the evaluation of antioxidant activity using DPPH and Frap assays. This perhaps can be explained by the presence of active compounds in this plant during the ripening period which participated in the antioxidant activity and this composition changes during the flowering period.

 

Figure 7. PPM activities of the roots of Juncus rigidus extracts during flowering and ripening period

 

The physiological stage affects the composition and content of polyphenols and biological activity⁵⁶. The distribution of secondary metabolites may change during plant development, perhaps related to the harsh climatic conditions of the plant’s usual habitat (hot temperature, high solar exposure, drought, salinity), which stimulate the biosynthesis of secondary metabolites such as polyphenols⁵⁷.

 

Cyclic voltammetry assay:

The Cyclic voltammetry assay is an effective method for the study of the electrochemical behavior of chemical substances. Using the obtained standard curve equation of the ascorbic acid (y = 119,46 x + 13,834 with R² = 0.9981) where y represents the anodic current density and x the standard concentration we can calculate the antioxidant capacity expressed in equivalent terms of ascorbic acid. The electrochemical data of the voltammograms above concerning anodic potential (Epa) and the intensity of the anodic current (Ipa) of each extract were presented in table 8.

 

Table 8. Electochemical data of the roots of Juncus rigidus extracts during flowering and ripening period

 

Figure 8 and figure 9 represented respectively cyclic voltammograms of the acetone and ethanol extract of the roots of Juncus rigidus during the flowering period and the acetone and ethanol extract during the ripening period. In all voltammograms a single oxidation peak was observed while the reduction peak does not exist, higher Ipa could be an indication of the higher antioxidant content of extract⁵⁸. Acetone extract during the ripening period has the highest anodic current density (8,6218 µA/cm²) which shows its highest antioxidant concentration compared to the other extract. This indication is very important and shows the efficiency of cyclic voltammetry.

 

Figure 8. Cyclic voltammogram of acetone extract (A) and ethanol extract (B) of the roots of the Juncus rigidus during flowering period                      

 

Figure 9. Cyclic voltammogram of acetone extract (A) and ethanol extract (B) of the roots of the Juncus rigidus during ripening period

Electrochemical properties of pure compounds, foods, and biological samples may be used for the evaluation of their reducing/antioxidant capacity since the electric oxidation potential has a conceptual relationship with the expected antioxidant capacity⁵⁹. Table 9 and figure 10 represent the ascorbic acid equivalent antioxidant capacities of the studied extracts of the roots of Juncus rigidus.

   

Table 9. antioxidant capacity of the roots of Juncus rigidus extracts during flowering and ripening period

 

Figure 10. Ascorbic acid antioxidant capacity (mg/g) of the roots of Juncus rigidus extracts during flowering and ripening period.

 

The obtained results showed that the acetone extract during the ripening period has the highest antioxidant capacity (47,07 ± 0,04 mg/g) compared to the other extracts. Ethanol extract during the flowering period was the lowest one (9,269 ± 0,01mg/g) and the most effective results are obtained during the ripening period compared to the flowering period. These results vary in relativity with those found by the DDPH, FRAP, and Phosphomolybdum assay.

 

CONCLUSION:

In this work of research, we have compared the contents of phenolics, flavonoids, and flavonols from the roots of Juncus rigidus and the evaluation of the antioxidant activity during the flowering and ripening period using two polar solvents acetone and ethanol for the extraction. No research has been done previously. All results indicated that acetone was very efficient in extracting active compounds from the roots of the Juncus rigidus plant and the best period where the composition of the plant in total phenolic contents, total flavonoid contents, and total flavonol contents was higher is the flowering period. The evaluation of the antioxidant activity with different methods during the two periods indicated that the higher antioxidant activity using DPPH, FRAP, phosphomolybdum assay, and cyclic voltammetry indicated that the strong antioxidant activity was obtained during the ripening period using acetone as a solvent for extraction this can be interpreted by the presence of the other active compounds during ripening period which had contributed in antioxidant activity so that the extraction yield was found higher during the ripening period.

 

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

 

ACKNOWLEDGMENTS:

The authors would like to thank Mr. Tliba Ali for his help in experiment analysis during this research.

 

REFERENCES:

1.     El-Shamy AI. Abdel-Razek AF. Nassar MI. Phytochemical review of Juncus L. genus (Fam. Juncaceae). Arabian Journal of Chemistry 2015; 8:614-623.

2.     Kovács A. Vasas A. Hohmann J. Natural phenanthrenes and their biological activity. Phytochemistry 2008; 69(5):1084-110.

3.     Ghareeb MA. Refahy LA. Saad AM. Ahmed WS. Chemical composition, antioxidant and anticancer activities of the essential oil from Eucalyptus citriodora (Hook.) leaves. Der Pharma Chemica. 2016; 8(1):192-200.

4.     Benarima A. Laouini SE. Ben Seghir B. Belaiche Y. Ouahrani MR. Optimization of Ultrasonic-Assisted Extraction of Phenolic Compounds from Moringa Oleifera Leaves using Response Surface Methodology. Asian Journal of Research in Chemistry. 2020; 13(5):307-311.

5.     Shlini P. Siddalinga Murthy KR. Extraction of Phenolics, Proteins and Antioxidant Activity from Defatted Tamarind Kernel Powder. Asian Journal of Research in Chemistry. 2011; 4(6): 936-941.

6.     Gopalasatheeskumar K. Ariharasiva Kumar G. Sengottuvel T. Sanish Devan V. Srividhya V. Quantification of Total Phenolic and Flavonoid content in leaves of Cucumis melo var agrestis using UV- spectrophotometer. Asian Journal of Research in Chemistry. 2019; 12(6):335-337.

7.     Rajurkar RM. Jain RG. Bedmohta PA. Khadbadi SS. Antioxidant Activity of Phenolic Extract from Ginger (Zingiber officinale Roscoe) Rhizome. Asian Journal of Research in Chemistry. 2009; 2(3):  260-261.

8.     Tiwari P. Estimation of Total Phenolics and Flavonoids and Antioxidant Potential of Amritarishta Prepared by Traditional and Modern Methods. Asian Journal of Research in Chemistry. 2013; 6(12): 1173-1178.

9.     Messaoudi A. Dekmouche M. Rahmani Z. Bensaci C. Comparative Analysis of Total Phenolics, Flavonoid content and Antioxidant profile of date palm (Phoenix dactylifera L.) with different watering water from Oued Soufin Algeria; Asian Journal of Research in Chemistry. 2020; 13(1): 28-32.

10.  Habchi A. Dekmouche A. Hamia C. Saidi M. Yousfi M. Bouguerra A. Extraction of Phenolic Compounds of six Algerian date (Phoenix dactylifera L.) cultivars from Ain-Saleh region, using Reflux method and Screening of Antioxidant Activity in vitro. Asian Journal of Research in Chemistry. 2021; 14(3):161-7.

11.  Zehiroglu C.  Beyza S. and  Sarikaya O. The importance of antioxidants and place in today’s scientific and technological studies. Journal of Food Science and Technology. 2019; 56(11):4757–4774.

12.  Isildak Ö. Yildiz I. Genc N. A new potentiometric PVC membrane sensor for the determination of DPPH radical scavenging activity of plant extracts. Food chemistry. 2021; 373: 131420.

13.  Thaipong K. Boonprakob U. Crosby K. Cisneros-Zevallos L. Byrne DH. Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. Journal of Food Composition and Analysis. 2006; 19: 669–675.

14.  Li R. Guo M. Zhang G. Xu X. Li K. Neuroprotection of nicotiflorin in permanent focal cerebralischemia and in neuronal cultures. Biological and Pharmaceutical Bulletin. 2007; 29: 1868-1872.

15.  Bahorun T. Gressier B. Trotin F. Brunet C. Dine T. Luyckx M. Vasseur J. Cazin M. Cazin JC. Pinkas M. Oxygen species scavenging activity of phenolic extracts from hawthorn fresh plant organs and pharmaceutical preparations. Arzneimittelforschung. 1996; 46(11):1086-9.

16.  Formagio ASN. Volobuff CRF. Santiago M. Cardoso CAL. Vieira MDS and Pereira ZE. Evaluation of antioxidant activity, total flavonoids, tannins and phenolic compounds in Psychotria leaf extracts. Antioxidants. 2014; 3: 745-757.   

17.  Burits M. Bucar F. Antioxidant activity of Nigella sativa essential oil. Phytotherapy Research. 2000; 14: 323-328.

18.  Cuendet M. Hostettmann K. Potterat O. Iridoidglucosides with free radical scavenging properties from Fagraea blumei. Helvitica Chimica Acta.1997; 80(4): 1144–1152.

19.  Que F. Mao L. Pan X. Antioxidant activities of five Chinese rice wines and the involvement of phenolic compounds. Food Research International. 2006; 39 (5):581–587.

20.  Amo-Mensah J. Darko G. Borquaye LS. Anti-inflammatory and antioxidant activities of the root and bark extracts of Vitex grandifolia (Verbanaceae). Scientific African. 2020; 10: e00586.

21.  Labed B. Bouhrira A. Benaissa K. Gherraf N. Radical scavenging and Antioxidant activities of three extracts of Lavandula coronopifolia rown in El-Hoggar, Tamanrasset, Algeria. Der Pharma Chemica. 2016; 8(9):132-139.

22.  Benzie IF. Strain JJ. The Ferric Reducing Ability of Plasma (FRAP) as a Measure of ‘‘Antioxidant Power’’: The FRAP Assay. Analytical Biochemistry. 1996; 239: 70–76.

23.  Fernandes RPP. Trindade MA. Tonin GF. Lima CG. Pugine SMP. Munekata PES. Lorenzo JM.  De Melo MP. Evaluation of antioxidant capacity of 13 plant extracts by three different methods: cluster analyses applied for selection of the natural extracts with higher antioxidant capacity to replace synthetic antioxidant in lamb burgers.  Journal of Food Science and Technology. 2016; 53(1): 451- 460.

24.  Amidi S. Majob F. Moghaddem AB. Tabib K. Kobarfard F. A simple electrochemical method for the rapid estimation of antioxidant potentials of some selected medicinal plants. Iranian journal of pharmaceutical research. 2012; 11 (1): 117-121.

25.  Guediri I. Boubekri C. Smara O. Lanez T. Total phenolic contents and determination of Antioxidant activity by DPPH, FRAP, and cyclic voltammetry of the fruit of Solanum nigrum (black nightshade) growing in the south of Algeria.  Asian Journal of Research in Chemistry. 2021; 14(1):47-55.

26.  López A. Rico M. Rivero A. Tangil MS. The effects of solvents on the phenolic contents and antioxidant activity of Stypocaulon scoparium algae extracts. Food Chemistry. 2011; 125: 1104–1109.

27.  Zhao H. Dong J. Lu J. Chen J. Li Y. Shan L. Lin Y. Fan W. Gu G. Effect of extraction solvent mixtures on antioxidant activity evaluation and their extraction capacity and selectivity for free phenolic compounds in Barley (Hordeum vulgare L.). Journal of Agricultural and Food Chemistry. 2006; 54: 7277–7286.

28.  Musa KH. Abdullah A. Jusoh K. Subramaniam V. Antioxidant Activity of Pink-Flesh Guava (Psidium guajava L.): Effect of Extraction Techniques and Solvents. Food Analytical Methods. 2011; 4: 100-107.

29.  Jacotet-Navarro M. Laguerre M. Fabiano-Tixier AS. Tenon M. Feuillère N. Bily A. Chemat F. What is the best ethanol-water ratio for the extraction of antioxidants from rosemary? Impact of the solvent on yield, composition, and activity of the extracts. Electrophoresis. 2018; 39:1946-1956.

30.  Truong D. Nguyen DH. Ta NTA. Bui AV. Do TH. Nguyen HC. Evaluation of the Use of Different Solvents for Phytochemical Constituents, Antioxidants, and In Vitro Anti-Inflammatory Activities of Severinia buxifolia. Journal of Food Quality. 2019; 1:1-9.

31.  Mollaei S. Hazrati S. Lotfizadeh VD. Dastan D. Asgharian P. Phytochemical variation and biological activities of Zosima absinthifolia during various stages of growth. International Journal of Food Properties, 2020, 23(1), 1556-1567.

32.  AlFaris NA. AlTamimi JZ. AlGhamdi FA. Albaridi NA. Alzaheb RA. Aljabryn DH. Aljahani AH. AlMousa LA. Total phenolic content in ripe date fruits (Phoenix dactylifera L.): A systematic review and meta-analysis. Saudi Journal of Biological Sciences.2021; 28:3566–3577.

33.  Herrera-Pool E. Ramos-Díaz AL. Lizardi-Jiménez MA. Pech-Cohuo S. Ayora-Talavera T. Cuevas-Bernardino JC. García-Cruz U. Pacheco N. Effect of solvent polarity on the ultrasound assisted extraction and antioxidant activity of phenolic compounds from habanero pepper leaves (Capsicum chinense) and its identification by UPLC-PDA-ESI-MS/MS. Ultrasonics Sonochemistry. 2021; 76: 105658.

34.  Ma YL. Sun P. Feng J. Yuan J. Wang Y. Shang YF. Niu XL. Yang SH. Wei ZJ. Solvent effect on phenolics and antioxidant activity of Huangshan Gongju (Dendranthema morifolium (Ramat) Tzvel. cv. Gongju) extract. Food and Chemical Toxicology. 2021; 147: 111875.

35.  Mokrani A. Madani K. Effect of solvent, time and temperature on the extraction of phenolic compounds and antioxidant capacity of peach (Prunus persica L.) fruit. Separation and Purification Technology. 2016; 162: 68–76.

36.  Ben Jalloul A. Chaar H. Saïdani Tounsi M. Abderrabba M. Variations in phenolic composition and antioxidant activities of Scabiosa maritima (Scabiosa atropurpurea sub. maritima L.) crude extracts and fractions according to growth stage and plant part. South African Journal of Botany. 2022; 146:703-714.

37.  Kobus-Cisowska J. Szczepaniak O. Szymanowska-Powałowska D. Piechocka J. Szulc P. Dziedziński M. Antioxidant potential of various solvent extract from Morus alba fruits and its major polyphenols composition. Ciência Rural, Santa Maria. 2020; (50)1: e20190371.

38.  Sunila AV. Murugan K. Variation in phenolics, flavonoids at different stages of fruit development of Pouteria campechiana (Kunth) baehni and its antioxidant activity. International Journal of Pharmacy and Pharmaceutical Sciences. 2017; 9(11): 70-75.

39.  Chepel V. Lisun V. Skrypnik L. Changes in the content of some groups of phenolic compounds and biological activity of extracts of various parts of heather (Calluna vulgaris (L.) Hull) at different growth stages. Plants. 2020; 9(8): 926.

40.  Barreca D. Trombetta D. SmeriglioA. Mandalari G. Romeo O. Felice MR. Gattuso G. Nabavi SM. Food flavonols: Nutraceuticals with complex health benefits and functionalities. Trends in Food Science and Technology. 2021; 117: 194–204.

41.  Zhong R. Miao L. Zhang H. Tan L. Zhao Y. Tu Y. Prieto MA. Simal-Gandara J. Chen L. He C. Cao H. Anti-inflammatory activity of flavonols via inhibiting MAPK and NF-κB signaling pathways in RAW264.7 macrophages. Current Research in Food Science. 2020; 5: 1176–1184.

42.  Brasileiro BG. Leite JPV. Casali VWD. Pizziolo VR. Coelho OGL. The influence of planting and harvesting times on the total phenolic content and antioxidant activity of Talinum triangulare (Jacq.) Willd. Acta Scientiarum. Agronomy. 2015; 37(2): 249-255.

43.  de Freitas Laiber Pascoal G. de Almeida Sousa Cruz MA. Pimentel de Abreu J. Santos MCB. Bernardes Fanaro G. Júnior MRM. Freitas Silva O. Moreira RFA. Cameron LC. Simões Larraz Ferreira M. Teodoro AJ. Evaluation of the antioxidant capacity, volatile composition and phenolic content of hybrid Vitis vinifera L. varieties sweet sapphire and sweet surprise. Food Chemistry. 2022; 366:130644.

44.  Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958; 181: 1199–1200.

45.  Belfar A. Bensaci C. Belguidoum M. Evaluation of antioxidant capacity by cyclic voltammetry of Phoenix dactylifera L. (date palm). Asian Journal of Research in Chemistry. 2022; 15(2):138-4.

46.  Ghimire BK. Seo JW. Kim SH. Ghimire B. Lee JG. Yu CW. Chung IM. Influence of harvesting time on phenolic and mineral profiles and their association with the antioxidant and cytotoxic effects of Atractylodes japonica Koidz. Agronomy. 2021; 11: 1327.

47.  Makarova K. Sajkowska-Kozielewicz JJ. Zawada K. Olchowik-Grabarek E. Ciach MA, Gogolewski K. Dobros N. Ciechowicz P. Freichels H. Gambin A. Harvest time afects antioxidant capacity, total polyphenol and favonoid content of Polish St John’s wort’s (Hypericum perforatum L.) fowers. Scientifc Reports, (2021) 11:3989.

48.  Ribeiro DA. Camilo CJ. de Fátima Alves Nonato C. Rodrigues FFG. Menezes IRA. Ribeiro-Filho J. Xiao J. de Almeida Souza MM. da Costa JGM. Influence of seasonal variation on phenolic content and in vitro antioxidant activity of Secondatia floribunda A. DC. (Apocynaceae). Food Chemistry. 2020; (15)315:126277.

49.  Pandey MM. Khatoon S. Rastogi S. Rawat AK. Determination of flavonoids, polyphenols and antioxidant activity of Tephrosia purpurea: a seasonal study. Journal of Integrative Medicine. 2016; 14(6):447-455.

50.  Do QD. Angkawijaya AE. Tran-Nguyen PL. Huynh LH. Soetaredjo FE. Ismadji S. Ju Y- H. 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.

51.  Rafińska K. Pomastowski P. Rudnicka J. Krakowska A. Maruśka A. Narkute M. Buszewski B. Effect of solvent and extraction technique on composition and biological activity of Lepidium sativum extracts. Food Chemistry. 2019; 15(289): 16-25.

52.  Sepahpour S. Selamat J. Abdul Manap MY. Khatib A. Abdull Razis AF. Comparative analysis of chemical composition, antioxidant activity and quantitative characterization of some phenolic compounds in selected herbs and spices in different solvent extraction systems. Molecules. 2018; 23(2): 402.

53.  Huyut Z. Beydemir Ş. Gülçin İ. Antioxidant and Antiradical Properties of Selected Flavonoids and Phenolic Compounds. Biochemistry Research International. 2017; 2017:7616791.

54.  Djemoui A. Djemoui D. Souli L. Souadia A. Gouamid M. The Antidiabetic, Antioxidant properties in vitro of Moringa oleifera flowers extracts grown in Sahara of Algeria. Asian Journal of Research in Chemistry. 2021; 14(3):173-8.

55.  Khan RA. Evaluation of flavonoids and diverse antioxidant activities of Sonchus arvensis. Chemistry Central Journal. 2012, 6(1):126.

56.  Medini F. Fellah H. Ksouri R. Abdelly C. Total phenolic, flavonoid and tannin contents and antioxidant and antimicrobial activities of organic extracts of shoots of the plant Limonium delicatulum. Journal of Taibah University for Science. 2014; 8(3): 216-224.

57.  del Baño MJ. Lorente J. Castillo J. Benavente-García O. del Río JA. Ortuño A. Quirin KW. Gerard D. Phenolic diterpenes, flavones, and rosmarinic acid distribution during the development of leaves, flowers, stems and roots of Rosmarinus officinalis. antioxidant activity. Journal of Agricultural and Food Chemistry. 2003; 51(15):4247-53.

58.  Amidi S. Mojab F. Bayandori Moghaddam A. Tabib K. Kobarfard F. A simple electrochemical method for the rapid estimation of antioxidant potentials of some selected medicinal plants. Iranian Journal of Pharmaceutical Research. 2012; 11(1):117-21.

59.  Magalhães LM. Segundo MA. Reis S. Lima JL. Methodological aspects about in vitro evaluation of antioxidant properties. Analytica Chimica Acta. 2008; 613(1):1-19.

 

 

 

Received on 20.10.2022                    Modified on 15.11.2022

Accepted on 01.12.2022                   ©AJRC All right reserved