EJBR2021v11i4art404 ISSN 2449-8955 European Journal of Biological Research Research Article European Journal of Biological Research 2021; 11(4): 404-416 DOI: http://dx.doi.org/10.5281/zenodo.5484499 Phytochemical profile, total phenolic content and antioxidant activity of ethanolic extract of fumitory (Fumaria capreolata L.) from Algeria Ismahene Sofiane*, Ratiba Seridi Plant Biology and Environment Laboratory, “Medicinal Plants” Axis, Biology Department, Faculty of Sciences, BADJI Mokhtar - Annaba University, Bp 12, 23000 Annaba, Algeria * Corresponding author: Phone: 0698 10 37 64/ 07 98 42 26 19, E-mail: sofiane-ismahene@hotmail.fr Received: 19 June 2021; Revised submission: 08 August 2021; Accepted: 02 September 2021 https://jbrodka.com/index.php/ejbr Copyright: © The Author(s) 2021. Licensee Joanna Bródka, Poland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/) ABSTRACT: Fumitory or Fumaria capreolata L. is a medicinal plant, spontaneous and widely distributed in North Africa, particularly in Algeria. It has been recognized for centuries for its therapeutic virtues, and it is used in traditional medicine in the treatment of hepatobiliary diseases, gastrointestinal disorders and in the treatment of skin diseases. The phytochemical screening carried out on the aerial part of the species F. capreolata L., revealed the richness of this plant in secondary metabolites, such as alkaloids, catechic tannins, sterols and terpenes. On the other hand, we noticed the absence of cardinolides, leuco-anthocyanins, quinones and starch in all parts of the plant. Quantitative spectrophotometric analysis allowed us to detect the levels of total polyphenols using the reagent of Folin-Ciocalteu, according to the results obtained we find that the species F. capreolata is rich in these compounds (14.27 ± 1.65 mg GAE/g). The evaluation of the antioxidant activity was carried out using the DPPH method, indicated that the ethanolic extract of F. capreolata L. showed significant antioxidant activity, with an IC50 = 0.27 mg/ml. And it also has a strong inhibitory activity of the coupled oxidation of linoleic acid and β-carotene, with a percentage of 88.46 ± 1.02% at a concentration of 0.5 mg/ml. In addition, the crude extract of F. capreolata L., also exhibits a good iron reduction capacity, with a maximum optical density of 0.349 at a concentration of 0.5 mg/ml. Keywords: Fumaria capreolata L.; Phytochemical screening; Antioxidant activity; DPPH; β-carotene; FRAP. 1. INTRODUCTION Fumitory, Fumaria capreolata L. is a medicinal plant belonging to the Papaveraceae family, endemic to the Edough peninsula in Seraidi (Annaba Province), in Algeria. Commonly called by the local Algerian population “hechichate el siban”. Fumitory is used in traditional Algerian medicine in hepatobiliary dysfunction, gastrointestinal disorders and for the treatment of skin pathologies. In addition, it has been reported that traditional medicine from many countries like Pakistan and India also use this herb as: cholagogue, diuretic, laxative, sedative, tonic and also considered useful to treat abdominal cramps, fever, diarrhea as well as syphilis and leprosy [1]. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 405 European Journal of Biological Research 2021; 11(4): 404-416 Medicinal plants of the genus Fumaria represent an inexhaustible source of natural antioxidants. Moreover, the number of studies carried out on these plants as well as antioxidants of plant origin undoubtedly reflects their importance in many areas, medicine or food. The anti-radical properties of these natural products are often linked to their ability to perpetrate stable radicals [2]. The objective of this study is the knowledge of Algerian natural resources in medicinal plants. In this context, the phytochemical composition, the content of phenolic compounds and the antioxidant activity in vitro of the species F. capreolata L., from the North-East of Algeria were studied. 2. MATERIALS AND METHODS 2.1. Plant material Aerial parts of the species Fumaria capreolata L. were collected in full bloom and fruiting, between the years 2014 and 2015, from Edough in Seraidi, (Annaba Province, in Northeastern of Algeria). The samples were taken manually and randomly, the botanical identification was made according to the flora of Quezel and Santa [3], and validated by Doctor Hamel. T, teacher-researcher in plant physiology at the Biology Department of Annaba University, Algeria. 2.2. Phytochemical screening The screening method by tube reactions allowed us to identify some major chemical families such as: alkaloids, flavonoids, quinones, sterols and terpenes. This is a qualitative analysis based on coloring and/or precipitation reaction compounds of the major chemical families present. This is carried out in the dry and/or fresh plants [4]. These preliminary tests were carried out according to the techniques of Solfo [5] and Harborne [6]. Table 1 indicates the different chemical groups sought and the specific reagents used. Table 1. Reagents used in the characterization of chemical groups. Chemical groups Reagents Reagent composition Positive results Alkaloids Draragendorf Nitrate of bismuth + acetic acid orange-red precipitate Mayer potassium iodide + mercury chloride yellowish precipitate Flavonoids Shinoda Ethanol 95° + HCl (N/2) + (Mg ou Zn) orange color, red or purple Steroids and Terpenoids Lieberman Bouchard Acetic anhydride + sulfuric acid purple coloring, blue or green Tannins FeCl3 1% FeCl3 1% dark blue, green or black coloring. Quinones Bornstraëgen / red coloring or violet Anthocyanins HCl 20% pink coloring, red-orange Saponosides Distilled water Foam Index (MI): positive test if IM > 100 2.3. Preparation of the ethanolic extract The ethanolic extract of F. capreolata L. was prepared according to the method of Rehman et al. [7]. 10 g of the powdered herbal drug (leaf, stem and flower) and 150ml of 90% ethanol were placed in a soxhlet apparatus (behr Labor) at 40°C for a period of 150 min (eleven extraction cycles). The extract was filtered and evaporated to dryness under reduced pressure with a Rotavap (Hei-Vap Ultimate, Heidolph), the latter is considered to be the crude extract. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 406 European Journal of Biological Research 2021; 11(4): 404-416 2.4. Thin layer chromatographic analysis (TLC) The analyzes by thin layer chromatography or (TLC) were performed on Silicagel aluminum plates 60 F254 (MARCHERY-NAGEL). The plates are developed in saturated glass vessels with the appropriate eluent. The mobile phase consists of a binary mixture of solvents. The solvent systems used are as follows (the proportions are given by volume and they are classified by increasing polarity): - nonpolar extracts: dichloromethane/methanol (9:1) - polar extracts: dichloromethane/methanol (9.5:0.5) (9.8:0.2) - hexane/ethyl acetate (4:6) (2:8) The TLCs are analyzed in visible light and under U.V. (254 and 356 nm), before and after revelation with appropriate reagents. Using reagents provides additional information about the type of a molecule (specific reagent cases). Three alkaloids were used as controls: the atropine, berberine and scopolamine, and a flavonoid: quercetin. The retention factors (Rf) of the spots resulting from the separation were calculated and compared to those of the controls, thus allowing the identification of the different compounds in the extract. Table 2. Main reagents used for (TLC) development. Reagents Substances revealed Method of preparation and use Sulphuric anisaldehyde (Deleu-Quettier 2000) Versatile reagent - Prepare a 0.5% solution of p-anisaldehyde in a mixture of CH3OH/Ac OH/H2SO4 (85:10:5). - Spray on the plate. After intense heating, the organic compounds appear as colored spots in daylight. Mounir Alkaloid revealer - Prepare 10 ml of the stock solution and 20 ml of acetic acid and make up to 100 ml with distilled water. - Spray on the plate: the alkaloids appear as orange-colored spots in daylight. 2.5. Determination of total phenolic compounds by colorimetry The method that we were able to adapt to our plant material, described by Juntachote et al. [8]. 0.5ml of the ethanolic extract of F. capreolata L. diluted in 5 ml of distilled water was mixed with 0.5 ml of the Folin-Ciocalteu reagent (FCR) in a test tube. Then 0.5 ml of 20% (w/v) anhydrous sodium carbonate solution (Na2CO3) was added to the mixture. After incubating the reaction mixture for one hour at room temperature in the dark, the absorbance is measured at 765 nm. The phenolic content of the extract was determined from the regression equation of the calibration range established with gallic acid. The results are expressed in mg gallic acid equivalent per gram of dry plant material (mg GAE/g). All the measurements are repeated 3 times. 2.6. Evaluation of antioxidant activity in vitro 2.6.1. DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging activity The experimental protocol followed to measure the scavenging activity of DPPH is that by Benhammou et al. [9]. The DPPH is dissolved in methanol to obtain a solution of 0.3 mM. In tubes 1 ml of methanol and 1 ml of the ethanolic extract (at different concentrations 1 mg/ml in methanol) are introduced and 2 ml of the methanolic solution with DPPH is added. After vortexing, the tubes are placed in the dark at room temperature for 30 minutes. The reading is taken by measuring the absorbance with a spectrophotometer (Perkin-Elmer UV/Vis Lambada 35) at 517 nm. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 407 European Journal of Biological Research 2021; 11(4): 404-416 The negative control is composed of 1 ml of the methanolic solution with DPPH and 2.5 ml of methanol. BHT, BHA and ascorbic acid have been used as standard synthetic antioxidants. The percentage of antioxidant activity was determined according to the following equation: % Anti-free radical activity = (Abs control - Abs sample / Abs control) × 100 The results are the mean of three separate measurements ± standard deviation. Calculation of IC50: IC50 or 50% inhibitory concentration is the concentration of the test sample necessary to reduce 50% of the free radical DPPH. 2.6.2. β-carotene bleaching method The experimental protocol followed is that of Ozsoy et al. [10]. The β-carotene/linoleic acid emulsion was prepared by dissolving 2 mg of β-carotene in 10 ml of chloroform, then 1 milliliter of this solution is mixed with 20 mg of purified linoleic acid and 200 mg of Tween 40, the chloroform was completely evaporated on a rotary evaporator at 40°C. And the residue obtained is taken up in 50 ml of water saturated with oxygen (H2O2), the resulting emulsion was stirred vigorously. Tubes containing 5 ml of this emulsion are prepared, for which 200 μl of a solution of the ethanolic extract of the plant studied or of reference antioxidant (BHA) at different concentrations are added. The mixture is stirred well and the absorbance reading at 470 nm is taken immediately against a blank, which contains the emulsion without the β-carotene. The covered tubes are placed in a water bath set at 50 ° C and the absorbance reading is taken after 120 minutes. A negative control is carried out in parallel, comprising 5 milliliters of the β-carotene emulsion and 200 μl of ethanol. The results obtained are expressed as a percentage inhibition of β-carotene discoloration using the following formula: Percent inhibition = [1 - (A₀ - At /A00 - A0t )] × 100 [10]. where: AA: Antioxidant activity; A°: Absorbance of the sample at t₀ At: Absorbance of the sample after 120 minutes of incubation A00: Absorbance of negative control at t₀ A0t: Absorbance of negative control after 120 minutes of incubation. 2.6.3. Ferric reducing antioxidant power (FRAP) The crude ethanolic extract of Fumaria capreolata L. diluted (1 ml) at different concentrations was mixed with 2.5ml of the phosphate buffer solution (0.2 M, pH 6.6) and 2.5 ml of potassium ferricyanide (K3Fe(CN)6) at 1%. The whole was incubated at 50°C for 20 min. Then, 2.5 ml of 10% trichloroacetic acid (TCA) was added to the mixture to stop the reaction, then the tubes are centrifuged for 10 min at 3000 rpm. Distilled water (2.5ml) and ferric chloride (FeCl3) (0.1%) were added to 2.5 ml of the supernatant. The absorbance reading was measured at 700 nm against a blank using a spectrophotometer (Perkin Elmer UV/Vis Lambada 35) [11]. Ascorbic acid was used as a positive control at the same chosen concentrations and under the same operating conditions as the samples. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 408 European Journal of Biological Research 2021; 11(4): 404-416 3. RESULTS AND DISCUSSION 3.1. Phytochemical screening of Fumaria capreolata L. The results of the chemical screening carried out on the infused, the macerated and the powder of Fumaria capreolata L. from Eastern Algeria are shown in Table 3. Table 3. The Phytochemical profile of the different organs of Fumaria capreolata L. Phytochemical families Fumitory Fumaria capreolata L. Flower Leaf Stem Root Alkaloids + + + + Flavonoids - - - - Sterols and terpenoids + + + + Tannins gallic + + + - catechetical - + + + Free quinones - - - - Anthocyanins + - + - Leuco anthocyanins + - - - Saponosides + + + + Starch - - - - Cardinolides - - - - Coumarins + - - - According to the results of the phytochemical screening carried out on the species F. capreolata L., we were able to detect different families of chemical compounds co-existing in this species by staining and precipitation reactions. Table 3 shows that the organs of the species Fumaria capreolata L. contain several chemical groups. We observed a significant presence of alkaloids, sterols, catechic tannins and saponins, in all organs of the plant. While coumarins could only be detected in the flowers. We noted a virtual absence of anthocyanins and leuco-anthocyanins in all organs, except in the flowers and stems of climbing fumitory where they are present. However, testing for free quinones, cardinolides, flavonoids and starch produced a negative inference for all organs of the plant. Much research has revealed the richness of European, Asian and African species of the genus Fumaria in different types of isoquinoleic alkaloids, in particular aporphine, protoberberine, protopine and benzophenanthridine. Also spiro-benzylisoquinoline alkaloids have been isolated such as fumaricin, fumarilin, fumaritin, fumarophycin, O- methylfumarophycin and parfumin [12-14]. Maiza-Benabdesselam and his collaborators [15] in Algeria identified the alkaloids contained in the methanolic extracts of the aerial part of two Algerian species: F. capreolata and F. bastardi by GC/MS (Gas Chromatography coupled with Mass Spectrometry). They showed the presence of a large number of alkaloids such as: stylopine, protopine, fumaritin, fumaricin, fumarophycin, fumarilin and fumarofin. Gupta and Rao [16] in a phytochemical study of the methanolic extract of the species Fumaria indica, observe four major chemical groups: alkaloids, flavonoids, sterols and saponins. Several authors have also reported the presence of tannins, tri-terpenoids, saponins and flavonoids in different parts of the plant [17-19]. These results confirm the presence of these different chemical compounds in the organs of our plant: Fumaria capreolata L. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 409 European Journal of Biological Research 2021; 11(4): 404-416 3.2. Yield of ethanolic extract of F. capreolata L. The results we obtained indicate that from 10 g of the powder of aerial parts of the plant Fumaria capreolata L. and 150 ml of ethanol and following evaporation to dryness of ethanol. We obtained an ethanolic extract considered to be the crude extract of blackish green color and with a semi-solid appearance. This extract may contain chlorophyll, polyphenols and other compounds. The yield is calculated on the total weight of dry and ground plant material, is expressed as a percentage. According to the results, the ethanolic extract of the plant studied has a low yield with a percentage of 8.4 ± 1.94%. This yield is higher than that obtained in another study carried out on the same species collected from the city of Constantine (Algeria), where the yield of the crude ethanolic extract is of the order of 2.5% [20]. Furthermore, the value we obtained in our study is consistent with the results obtained in other work carried out on species of the same genus, where the ethanolic extracts of Fumaria officinalis and F. parviflora gave similar yields with the values of the order of 8% and 7.4%, respectively [21, 22]. However, Mohajerani et al in 2019, mentioned that the ethanolic (80%) extract of the species Fumaria vaillantii L. has a very high yield with a percentage of 20.3% [23]. While other researchers have reported a much lower yield, with a percentage of 11% and 10.2%, for ethanolic extracts of F. officinalis and F. indica [24, 25]. The difference in yield noted in our study may be due to the chemical composition which differs from one species to another, the possible content of active ingredients, the plant material to be extracted, the nature of the solvent used and the extraction technique used without forgetting the nature and composition of the soil [26]. 3.3. Thin layer chromatographic analysis (TLC) In the analysis of plant extracts by thin layer chromatography, each substance is characterised by its fluorescence under UV light (254 nm and 365 nm), its "Rf" and its color after development with the appropriate chemical developer. According to the results, we observe nine (9) spots with different colors and migration distances (Rf) under both wavelengths for the ethanolic extract of F. capreolata L. TLC plates developed in the sulphuric anisaldehyde (followed by heating) showed colored spots (green, blue and black spots), indicating the presence of organic compounds. On the other hand, the appearance of certain orange-yellow colored spots after the revelation of the plates by the Mounir reagent indicates the presence of alkaloids. Figure 1. Chromatograms of the ethanolic extract of F. capreolata L. after development. TLC showed the probability of the presence of isoquinoline alkaloid such as berberine (Rf = 0.80) and the absence of the atropine and scopolamine in the ethanolic extract of the studied plant. Also a large group of Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 410 European Journal of Biological Research 2021; 11(4): 404-416 phenolic compounds was detected. These results are consistent with those of Naz and co-workers in 2013 and Guna in 2017 in their studies on related species: Fumaria parviflora and Fumaria indica [27, 28]. 3.4. Total phenolic content of ethanolic extract of Fumaria capreolata L. The family of phenolic compounds includes a large number of secondary metabolic products which differ in their structures and reactivity. These compounds have been of great interest in recent years due to their beneficial effects on human health, their powerful antioxidant properties and their credible effects on the prevention of various diseases associated with oxidative stress [29]. Figure 2. Gallic acid calibration curve for the determination of total polyphenols. The content of phenolic compounds in the ethanolic extract of the species F. capreolata L. is 14.27 ± 1.65 mg GAE/g. While, the ethanolic extract of Romanian F. capreolata contains the highest amount of total phenolics (18.56 mg GAE/g d.w.) [30]. In addition, Ivan et al. [31], studied the levels of phenolic compounds in ethanolic extracts of five species of the genus Fumaria: F. officinalis, F. thuretii, F. kralikii, F. rostellata and F. schrammii. They showed that the polyphenol content in these species is between 20.20 ± 0.29 mg GAE/g (in the species F. thuretii) and 30.30 ± 0.31 mg GAE/g (in the species F. officinalis). These values are significantly higher than what we noted in our study on the species F. capreolata L. Significantly, the lowest values of phenolic compounds were recorded by Orhan et al. [32], in their study on four species of the genus Fumaria collected from Turkey: F. cilicica, F. densiflora, F. kralikii and F. parviflora. The values obtained varied between 0.05 and 0.09 mg GAE/g of dry extract. According to these results, we find that the species F. capreolata L. collected from Algeria is richer in phenolic compounds. The variation noted in in the quantity of polyphenols from one extract to another and from one species to another is probably due to several parameters such as: the operating conditions of the extraction, the nature and the polarity of the solvent used. These variations are also considerable depending on the variety, the physiological stage of the plant and the nature of the plant tissues [33]. 3.5. Antioxidant activity 3.5.1. DPPH radical scavenging activity The anti-free radical activity of extract was evaluated by the DPPH spectrometry method. The DPPH molecule is a free radical, dark purple in color, characterized by an absorption band between 515-520 nm. The DPPH free Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 411 European Journal of Biological Research 2021; 11(4): 404-416 radical scavenging assay is based on the reduction of the latter when mixed with an antioxidant such as polyphenols, which leads to a loss of its violet color which turns pale yellow and to a reduction in its absorption at 520 nm [29]. Figure 3 reports the percentages of inhibition obtained from the ethanolic extracts of F. capreolata L., Compared to that of the positive controls used (BHA, BHT and ascorbic acid). Figure 3. Anti-free radical activity of the ethanolic extract of Fumaria capreolata L. Each value represents the mean of three tests ± SD. Figure 3 shows that the percentages of inhibition are important at different concentrations; which increase in anti-free radical activity proportional to the increase in the concentration of the extract tested. At a concentration of 0.5 mg/ml the ethanolic extract of Fumaria capreolata L. shows significant anti-free radical activity with a high DPPH radical scavenging power (72.35 ± 0.27%). Table 4. IC50 found in the extract of the plant studied. Extracts IC50 expressed in mg/ml Ethanolic extract of F. capreolata L. 0.0300 BHT 0.0078 BHA 0.0058 Ascorbic acid 0.0071 From the results shown in the Table 4, we note that the three positive controls used have a potent anti-free radical activity and superior to that of the extract of the plant studied. Indeed, BHA is the most active with an IC50 = 0.0058 mg/ml, then ascorbic acid, then BHT (0.0071 mg/ml and 0.0078 mg/ml). We also note that the ethanolic extract of F. capreolata L. showed a still high IC50 value (0.030 mg/ml). Much research describes the antioxidant activity by trapping the free radical DPPH of species of the genus Fumaria. Previously, Bribi et al. [34] evaluated the antioxidant activity of the extract of total alkaloids of the same species Fumaria capreolata L., by the DPPH radical scavenging method in a concentration range between 0 and 800 µg/ml. The strong anti-free radical effect of the extract tested was estimated at 68.31 ± 0.35% at a concentration of 100 µg/ml. And the IC50 values found in the tested extract and the BHA were in the order of 28.87 µg/ml and 8.21 µg/ml. This result is clearly superior to that which we obtained in our study, where the ethanolic extract of the aerial part of the species Fumaria capreolata L. collected from the Edough region (Annaba, Algeria), seems to have anti-radical activity with a DPPH radical scavenging power of around 72.35 ± 0.05% at a concentration of 0.5 mg/ml. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 412 European Journal of Biological Research 2021; 11(4): 404-416 In the same context, Maiza Benabdesselam and these collaborators [15], studied the antioxidant activity of alkaloids extracts of the two species Fumaria capreolata L. and Fumaria bastardii native to Algeria. The high inhibitory effect of the free radical DPPH was exerted by the extract of the total alkaloids of the species F. bastardii with a percentage of 86%, followed by the extract of the total alkaloids of the species F. capreolata L. with a percentage inhibition of 45.6%, only at a concentration of 50 µg/ml. Several other studies have demonstrated the anti-free radical activity of the ethanolic extract of F. indica by this same method, and each time the extract has shown a significant inhibitory power of the order of 61.8% [19], and with an IC50 of 11 mg/ml [25]. In the work of Orhan et al. [34], the antioxidant activity of several types of extracts from four species of the genus Fumaria: F. cilicica, F. densiflora, F. kralikii and F. parviflora was studied by several methods. The results obtained show that the extracts tested have a significant DPPH radical scavenging activity at the concentrations of 250, 500 and 1000 μg/ml. And the fraction of ethyl acetate and dichloromethane of the species F. cilicica exhibits the highest activity with a percentage of inhibition of the order of 76.16 ± 0.12% and 51.86% respectively, at a concentration of 1000 µg/ml. Theremore, Ivan et al. [31], observed that the ethyl acetate extract of the species F. vaillantii has the highest antioxidant activity compared to the rest of the extracts tested, with a percentage inhibition of the DPPH radical of 83.41%. This result joins that of Moghaddam et al. [35], who noted that the highest antioxidant power of the same species was observed at the vegetative stage, with an IC50 value of the order of 1217.85 ± 1.02. These authors reported that this activity is probably due to the presence of phenolic compounds and flavonoids. This antioxidant property of the species F. capreolata L. may be due to protopine, which is an isoquinoline alkaloid present in the ethanolic extract of this plant as a major compound. The latter is endowed with several biological activities. Species of this genus also contain a number of fatty acids with an antioxidant effect, such as: linoleic acid, oleic acid, palmitic acid and myristic acid [30]. 3.5.2. β-carotene bleaching method The antioxidant power of our extract has also been tested by the β-carotene bleaching method. The oxidation of linoleic acid generates peroxide radicals, and conjugated diene hydro peroxides. This test is based on the fact that these freed radicals will subsequently oxidize the highly unsaturated β-carotene, which loses its double bonds, thus causing the disappearance of its red color, which is measured spectrophotometrically at a wavelength λ = 490 nm. However, the presence of an antioxidant could neutralize free radicals derived from linoleic acid, and therefore prevents the oxidation and bleaching of β-carotene [37]. From our results, we clearly notice that the ethanolic extract of the studied plant and BHA exert a powerful inhibitory effect on the oxidation of β-carotene, after 120 minutes of incubation. The ethanolic extract of F. capreolata L. shows the greatest inhibitory activity of the coupled oxidation of linoleic acid and β-carotene with a percentage of 88.46 ± 1.02% at a concentration of 0.5 mg/ml, followed by BHA with a percentage of 82.69 ± 0.03%. According to the literature, two species of the genus Fumaria native to Algeria were investigated for their ability to inhibit the peroxidation of linoleic acid. Maiza-Benabdesselam et al. [15] studied the antioxidant activity extracts of the total alkaloids of F. capreolata L. and F. bastardii L. The alkaloids extracts of both plants expressed strong antioxidant activity; however, the activity of F. bastardii extract was more potent than of F. capreolata L. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 413 European Journal of Biological Research 2021; 11(4): 404-416 Figure 4. The antioxidant power of the ethanolic extract of Fumaria capreolata L. tested by β-carotene decoloration method. At a concentration of 500 μg/ml, the two extracts tested showed a percentage of 65 and 67.8% inhibition of the peroxidation of linoleic acid, for the species F. capreolata L. and F. bastardii respectively. On the other hand, at the same concentration the antioxidant butylated hydroxyanisole (BHA), quercetin, and caffeine have an inhibition rate of 80, 56.2 and 64.3%, respectively. 3.5.3. Ferric reducing antioxidant power (FRAP) The FRAP method is a simple, inexpensive and robust spectrophotometric technique. It is based on the ability of polyphenols to reduce ferric iron Fe³⁺ to ferrous iron Fe²⁺ [40]. From the results obtained, we note that the increase in ferric iron reducing absorbances is proportional to the concentrations used. The crude extract of F. capreolata L. expressed a very low reducing power, with observed values of optical densities not exceeding 1 (OD = 0.349 ± 0.0062) at a concentration of 0.5 mg/ml. From the graphs shown in figure 3, we can clearly observe the low capacity of the crude extract of F. capreolata to reduce iron by comparing the latter with the reducing power of ascorbic acid which is of the order from 2.52 ± 0.0052. This potential is related to the nature of the reducing substances existing in the extract tested. Figure 5. Reducing power of the ethanolic extract of the plant studied and of the ascorbic acid tested by the FRAP method. Each value represents the average of three trials. Sofiane & Seridi Phytochemical profile, total phenolics and antioxidant activity of Fumaria capreolata 414 European Journal of Biological Research 2021; 11(4): 404-416 A compound's reducing capacity can serve as an indicator of its antioxidant potential. The presence of reducing agents (such as antioxidants) causes the conversion of Fe3+ ferricyanide complex in the ferrous form Fe2+. Although iron is essential for oxygen transport for respiration and enzyme activity, it is a reactive metal that catalyzes oxidative damage in living tissues and cells [41]. The reducing activity of the ethanolic extract of the plant Fumaria capreolata L. from the Edough region was moderate and significantly lower than that of ascorbic acid, with an optical density of 0.349 ± 0.03 at a concentration of 0.5 mg/ml. Our results join those obtained by Maiza-Benabdesselam et al. [15], where the extracts of the total alkaloids of F. bastardii and F. capreolata showed a low activity for the reduction of iron compared to the standards used, in the following order: quercetin > BHA > gallic acid followed by the extracts of the total alkaloids of F. bastardii and F. capreolata. In another study, the best reducing capacity of the extract of the total alkaloids of F. capreolata was obtained at a concentration of 800 µg / ml with an optical density of around 0.57 ± 0.005 [33]. On the other hand, the ethanolic extract and the fractions of four plants belonging to the genus Fumaria from Turkey were tested for their reducing capacity. All of the extracts tested exerted a low reducing capacity compared with the positive control. And the greatest reducing activity was obtained from the ethanolic extract of Fumaria kralikii with an absorbance of 0.390 ± 0.04 at a concentration of 1000 µg/ml [37]. Further, the antioxidant activity of F. vaillantii extracts reported by FRAP assay, demonstrates that the reducing power of BHT (585.91 Fe2+/mg extract) was significantly higher than vegetative, budding and flowering stages (359.48 and 248.87 μmol Fe2+ per mg EO, respectively) [38]. Generally, the variation in the reducing activity is attributed to the chemical composition of the extracts tested. However, it may be due to one of the majority constituents or to other minority constituents or also to a synergy between them. 4. CONCLUSIONS We can conclude that, the plant that we studied as well as various other species of the same genus, were markedly different with regard to their phytochemical composition, in particular alkaloids and phenolic compounds. As a result, they also differ in their anti-oxidant activities. However, we noted that the antioxidant properties of the species Fumaria capreolata L from Edough region (Annaba Province, Algeria), correlated with their phytochemical composition. Authors' Contributions: SI: collecting plant samples, extraction, determination of total phenolic content, studied antioxidant activity of the plant and wrote the manuscript. SR: Supervised the findings of this work. All authors are read and approved the final manuscript. Conflict of Interest: The author has no conflict of interest to declare. REFERENCES 1. Gilani AH, Bashir S, Janbaz KH, Khan A. Pharmacological basis for the use of Fumaria indica in constipation and diarrhea. J Ethnopharmacol. 2005; 3: 585-589. 2. 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