INTRODUCTION:

The significant role of oxidation in the bodies and in nutrients has been widely acknowledged and it is necessary for cell’s functioning. One unintended result of this dependency is the formation of free radicals as well as other reactive oxygen species, causing oxidative alterations¹. Increasing data suggests that these species have a function in various conventional in vivo regulation systems. Excessive amounts of free radicals may overpower defensive enzymes like catalase, superoxide dismutase and peroxidase. This may result deadly impacts on organisms through oxidizing cellular proteins, membrane lipids, DNA, and enzymes, ultimately halting cell respiration².

Moreover, reactive oxygen species appear to modify cell signaling processes in previously unknown manners. An antioxidant is a substance which, even in little amounts, has the ability to inhibit or significantly slow down the process of oxidation in materials that are subject to oxidation³. It can also delay or prevent the oxidation of substances when exist in low quantities comparing to the oxidizable substrate. In addition, antioxidants possess the ability to regulate or counteract the process of lipid or biomolecule oxidation by scavenging reactive oxygen species. The action of different antioxidants serves as a means of protection against the harmful consequences of excessive oxidation, and the need of assessing antioxidant activity is widely acknowledged^4,5.

A natural state of balanced dynamic presents between the creation of free radicals in our bodies and the existence of antioxidants that shield the organism from detrimental effects. The quantity of antioxidant compounds available under typical physiological conditions is often insufficient to counteract the effects of free radicals. Consequently, there is an increasing fascination in the food industry and in preventive medicine regarding the creation of "natural antioxidants" derived from plant⁶. As a result, plants having antioxidant capabilities are becoming increasingly popular across the world.

Scabiosa is a taxonomically intricate genus consisting of several species that are distributed over the Mediterranean Basin, southern Africa and Asia⁷. Scabiosa genera are recognized for their medical properties, and studies on their phytochemical composition have shown the presence of intriguing secondary metabolites, among them have demonstrated interest as medical treatments⁸. Numerous reports have shown that Scabiosa species in the Mediterranean area possess a wide range of biopotential and therapeutic benefits. These species have demonstrated many biological benefits, notably antioxidant, anti-inflammatory and antibacterial activities⁹. Scabiosa atropurpurea is a yearly plant that forms clusters of leaves at the base and has stems with leaves, attaining an altitude of 20-60 cm. This genus may be identified by its unique bluish-lilac blossoms with peculiar fruit form^10,11.

To our understanding, the antioxidant properties of the above-ground portion of S. atropurpurea have not been investigated utilizing a comparison between aqueous and methanolic extracts. Hence, this research investigation seeks to analyze the antioxidant efficacy of aqueous and methanolic extracts from S. atropurpurea, a plant species found in the Algerian flora, utilizing different techniques and assays.

MATERIALS AND METHODS:

Plant species selection

The plant samples were gathered locally in Chaabia region of Ouricia, Setif, located in northeastern Algeria during the blossom and fruiting season of may to july (2021). The plant was identified under voucher specimen (020/DBEV/UFA/22). The entire aerial portion was completely cleansed, dehydrated in a shaded area, powdered, and thereafter kept for testing purposes.

Extractions procedures:

Preparation of the decocted extract:

The aqueous extract (AE) was obtained by boiled 100g of dehydrated aerial part in one liter of purified water for a duration of 20 minutes¹². Subsequently, the mixture underwent filtration using muslin cloth and was subjected to centrifugation at 3000 rpm for a duration of 15 minutes. The extract’s solution was gathered and evaporated to dryness. The resulting dry extract was then kept at a temperature of 4°C.

Preparation of the crude methanolic extract:

The complete above-ground portion of scabiosa atropurpurea weighing 4kg was defatted by 15L of petroleum ether. Subsequently, the plant was subjected to air drying and then underwent maceration using 15 liters of 100% methanol. The crude methanolic extract (CrE) was evaporated and the resultant concentrated extract was preserved at 4°C¹³.

Qualitative phytochemical screening:

Analysis of plant compounds using qualitative methods to identify the presence or absence of certain chemical constituents. The samples were analyzed to determine the presence of bioactive components including polyphenols, flavonoids, tannins, coumarins, quinones, terpenoids, and anthraquinones adopting the procedures described by^14,15

Quantification phytochemical estimation:

Total phenolic content:

Polyphenol tenor was conducted employing Folin-Ciocalteu (FC) procedure^16,17. 0.1 mL of samples was combined with 0.5 mL (FC) reagent. 0.4 mL of sodium carbonate (7.5%) was then introduced. Following two hours of incubation, the spectrophotometric measurement was estimated at 765 nm and the findings were represented as ug of gallic acid equivalents per mg of extract (ug GAE/mg DE).

Overall flavonoid tenor:

Aluminum chloride reagent (ACR) was utilized to calculate the total flavonoid amount¹². 0.5 mL of samples was combined with 0.5 mL ACR (2%), and the resulting combination was allowed to incubate for ten minutes. The spectrophotometric analysis conducted at 430 nm. The findings were expressed as ug of quercetin equivalent per mg of dried extract (µg QE/mg DE).

Condensed tannins content:

The condensed tannins tenor has been performed using a modifying vanillin procedure¹⁴. 250 μL of samples was combined with 375 μL of vanillin reagent (4%). Later, 187.5 μL of HCl was added. The resulting blend has been incubated for twenty minutes. The spectrophotometric measurement was conducted at 550 nm. The obtained data were reported in ug of catechin equivalent per mg of dry extract (μg CE/mg DE).

Evaluation of the anti-oxidant property:

DPPH radical scavenging activity:

Examining samples' capacity to remove free radicals has been achieved via measuring a decreased absorption (at 517 nm) of a DPPH reagent¹⁸. 50 μL of samples' varied doses or butylated hydroxytoluene (BHT) as a reference were blended with 1.25 mL of a DPPH reagent (0.004 %). The absorption was recorded at 517 nm over 30 min of incubating in the absence of light.

Iron chelating assay:

The assessment of ferrous iron chelation of samples has been performed through a spectrophotometric test¹⁹. 50 µL of Fecl2 (0.6 mM) and 450 µL of methanol were combined with 250 µL of extracts at various concentrations. 50 µL of ferrozine (5 mM) was introduced after five minutes, and the mixture was let to interreact for ten minutes. A reading of results was estimated at 562 nm. An IC50 value was calculated, which is the amount of products that produce 50% of the maximal neutralizing efficiency.

Reducing power assay:

This assay is employed to assess the reduction capacity of samples¹⁹. 400 μL of either extract or ascorbic acid was blended with a similar amount of phosphate buffer (0.2 M, pH = 6.6) and potassium ferricyanide (1%) and rest for 20 minutes at 50 °C. A 400 μL of 10% TCA was introduced and the produced mix was subjected to centrifugation at 3000 rpm during ten minutes, then combined with purified water (400 μL) and 80 μL of 0.1% ferric chloride. Following ten minutes of incubating, the formed product was observed at 700 nm.

Phosphomolybdate assay:

This method is used to evaluate the total antioxidant effect^20,21. One milliliter of combination consists of (28 mM sodium phosphate, 4 mM ammonium molybdate and 0.6 M H2SO4) has been blended with 100 μL of samples. The resulting mixture rest during 90 minutes at 95 °C. Next, the results were determined at 695 nm. The outcomes were determined as μg equivalents of ascorbic acid per mg of extract (μg EAA/mg extract).

Statistical analysis:

Statistical analysis implied Student’s and one-way ANOVA tests, coupled with Dunnett and Tukey tests. The results were provided in triplicate, with the mean value accompanied by the standard deviation (SD). The study was conducted using GraphPad Prism-8. Variation defined Significant at a p-value less than 0.05. An IC₅₀ was calculated by linear regression.

RESULTS:

Phytochemical screening:

The extracts of S. atropurpurea comprised various phytochemical compounds, as documented in Table 1. The outcomes demonstrated the existence of polyphenols, flavonoids, free quinones, tannins, saponins, terpenoids, anthraquinones, coumarins and reducing sugars in aqueous extract. However, CrE contains all phytochemical compounds except coumarins. This phytochemical analysis reveals the abundance of secondary metabolites in both extracts.

Table 1: Phytochemical screening of AE and CrE extracts from S. atropurpurea.

Phytochemical compounds	AE	CrE
Polyphenols	+	+
Flavonoids	+	+
Tannins	+	+
Terpenoids	+	+
Free quinones	+	+
Anthraquinones	+	+
Coumarins	+	-
Saponins	+	+
Reducing sugar	+	+

Extraction yields and determination of bioactive compounds:

The outcomes of extraction are illustrated in Table 2. The yield percent of CrE (13.8%) was higher than AE (9.06%). The overall phenolic amount, flavonoids, and condensed tannins are presented in Table 2. The findings of the quantified phytochemical assessment suggest that CrE exhibited a significant level of TPC and TFC with 114.13±0.92µg GAE/mg DE and 100.57±0.93 µg QE/mg DE, respectively. However, the highest CT content was recorded by AE with 41.04±0.64 µg CE/mg DE.

Table 2: Extraction yields and total phenolic, flavonoids and condensed tannins content of S. atropurpurea extracts.

Extracts

Yields %

TPC (µg GAE/

mg DE)^a

TFC (µg QE/

mg DE)^b

CT (µg CE/

mg DE)^c

9.06

81.6±1.6

45.33±0.36

41.04±0.64

CrE

13.8

114.13±0.92

100.57±0.93

26.22±1.11

Data are represented as mean ±SD (n=3). ^aGallic acid equivalent/mg of dried extract, ^bQuercetin equivalent/mg of dried extract, ^cCatechin equivalent/mg of dried extract.

In vitro antioxidant capacities:

The DPPH scavenging activity of the AE and CrE produced from S. atropurpurea was expressed using the inhibitory concentration (IC50), and the results appear in Table 3. Both extracts demonstrated a high eliminating efficacy compared to BHT. It is noteworthy that the significant chelator effect of AE (0.16±0.03mg/ml) compared to the EDTA as a reference (0.01 ± 0.00 mg/ml), whereas CrE exhibited the lowest effect (0.64 ± 0.01 mg/ml). The extracts' reducing power effect was evaluated by measuring A0.5 (mg/mL), a dose corresponds to the absorbance at 0.50 nm. Despite the ferrous reducing capacity of vitamin C (0.01 ± 0.00 mg/mL), AE and CrE demonstrated significant ability. Furthermore, it is essential to mention the superior reducing ability of AE compared to CrE (0.08 ± 0.003 mg/mL) (Table 3). The total antioxidant ability was assessed employing the phosphomolybdate test. AE and CrE showed almost a similar potency with 0.2±0.01 and 0.19±0.01 µg EAA/mg E, respectively.

Table 3: Antioxidant capacities of S. atropurpurea.

Extracts/ standards (mg/mL)	IC₅₀ (mg/ mL)		A_0.5 (mg/ mL)	µg EAA/ mg E
Extracts/ standards (mg/mL)	DPPH	Chelating assay	Reducing power assay	Phosphomoly- bdate assay
AE	0.06±0.00^**	0.16±0.03^***	0.08 ±0.00^***	0.2±0.01^ns
CrE	0.06 ± 0.02^**	0.64 ± 0.01^***	0.13 ± 0.01^***	0.19±0.01^ns
BHT	0.014 ± 0.00	--	--	--
EDTA	--	0.01 ± 0.00	--	--
Ascorbic acid	--	--	0.01 ± 0.00	--

Data are represented as mean ±SD of three measurements compared to standards. ns: no significant difference, ** p<0.01, *** p<0.001. BHT: Butylated hydroxytoluene, EDTA: Ethylenediaminetetraacetic acid

DISCUSSION:

For millennia, traditional medicine has utilized herbs to cure various ailments among different communities worldwide. The medical effectiveness can be related to the abundant collection of diverse bioactive components found in plants²². This research examines the phytochemical composition and antioxidant properties of S. atropurpurea utilizing several experimental techniques. The results indicated that the yields of the extracts varied, and this variation may be explained by taking into account the extraction technique used, the selection of solvents, and the particular circumstances within the extraction process²³. The phytochemical screening of S. atropurpurea revealed the existence of several secondary metabolites with established biological activity, involving flavonoids, polyphenols, free quinones, tannins, terpenoids, anthraquinones, saponins, coumarins, and reducing sugars. Multiple investigations have verified the abundance of phytochemical substances in scabiosa species, which have advantageous biological effects²⁴. Multiple studies on scabiosa species have conclusively demonstrated the presence of diverse bioactive chemicals S. atropurpurea²⁵, and S. comosa²⁶. Flavonoids were

isolated from S. caucasica²⁷, S. stellata²⁸ and S. atropurpurea²⁹. Terpenoids and irridoid glucosides were detected in S. stellata³⁰, S. tschilliensis³¹, S. atropurpurea subsp. Maritima^32,33 and S. arenaria³⁴. Coumarins were identified in S. hymettia³⁵. The amount of polyphenols, flavonoids, and condensed tannins was measured using spectrophotometry. The findings demonstrated a high level of these compounds, particularly when compared to other scabiosa species such as S. arenaria³⁴ and S. stellata³⁶. The choice of solvent for extraction and its polarity can have an impact on the phenolics content and the characteristics of the extracted molecules²¹.

Antioxidants are highly regarded for their significant health benefits and crucial role in preventing against diseases and the evaluation of antioxidant capacity should be conducted using multiple approaches³⁷. This study utilizes different in vitro techniques to assess the antioxidant properties of S. atropurpurea, including scavenging of free radicals, ferric reducing power, metal chelating, and the phosphomolybdate method. DPPH is a highly stable free radical that is commonly used to study the capability of natural products to scavenge free radicals. It is also the favored approach assessing the ability to remove free radicals of novel medications because of its simplicity and effectiveness and rapidity³⁸. The scavenging effect seen in AE and CrE was superior to that of the ethanolic extract of S. atropurpurea (IC50 0.1383 ± 0.0789 mg mL-1)24 and the methanolic extract of S. stellata (IC50=86 µg mL-1)³⁶. The antiradical action of S. atropurpurea extracts is attributed to their high content of polyphenols, which are well-known for their capacity to eliminate free radicals by donating hydrogen. Also, the most significant mechanisms of polyphenols are estimated to be free radical scavenging, in which they can interrupt the free radical chain reaction, and eradication of free radical formation through regulating enzyme activity³⁹. The scavenging effect is influenced by the presence and positioning of hydroxyl and carboxyl groups on the molecules of polyphenols. In addition, the metal chelation characteristic is used as a sign of antioxidant potential. It has been studied in conjunction with other antioxidant tests for various antioxidants and extracts⁴⁰. Antioxidants are shown to effectively bind iron due to their functional groups that work as metal binders⁴¹. The chelating effect values of S. stellata were inferior to our results³⁶. Moreover, the presence of polyphenols in extracts enables them to bind to metal ions, such as iron, thereby increasing their antioxidant properties by inhibiting redox-active transition metals from supporting free radical formation of free radicals⁴². Furthermore, one of the main characteristics of flavonoids is their antioxidant activity, plus their capability to chelate transition metal ions⁴³. Conversely, the reduction of antioxidant power assay is a widely used and dependable method for assessing antioxidant capacity⁴⁴. Furthermore, compounds possessing reduction potential react with potassium ferricyanide to produce potassium ferrocyanide. This compound then interacts with FeCl3, resulting in the creation of a highly concentrated blue complex with a maximum absorbance at 700 nm⁴⁵. Among extracts, AE revealed the greater reducing effect with 0.08 ± 0.00 mg mL^-1, this activity was higher than those of n-butanol fraction of flowers, stem and leaves from S. arenaria and lower than the fruits’ butanolic fraction³⁴. In fact, the features of components that reduce ferric ions are attributed to their ability to transport electrons, which is an important characteristic indicating their antioxidant capacity⁴⁶. A spectrophotometric test was devised to accurately measure the total antioxidant capacity by utilizing the phosphomolybdenum method. This approach relies on the conversion of molybdenum (IV) to molybdenum (V) by the plant samples, causing the creation of a phosphate/molybdenum (V) complex under acidic circumstances. This complex exhibits a green color its highest absorbance at 695 nm²¹. The phosphomolybdate assay has been documented as a reliable technique for assessing the overall antioxidant capabilities of different plant extracts⁴⁷. Our investigation found that both AE and CrE exhibited a suppressive effect on the molybdenum (IV), indicating a high antioxidant capability. The overall antioxidant capacity of S. atropurpurea may be related to the existence of flavonoids and phenolics in the plant's extracts.

The findings of this work confirm the conventional use of S. atropurpurea for treating various ailments and show that different extracts containing distinct secondary metabolites possess strong antioxidant properties and benefits. Typically, the antioxidant properties of plants are mainly attributed to the existence of polyphenols and may be influenced by other substances such as coumarins and flavonoids⁴⁸. Numerous investigations have demonstrated that plant polyphenols can serve as antioxidants, safeguarding against diseases caused by oxidative stress⁴⁹. Current studies have demonstrated that plants contain numerous bioactive chemicals that exhibit potent antioxidant actions. The antioxidant properties exhibited by our plant extracts can be related to the existence of polyphenols, flavonoids and tannins. The extracts exhibit properties such as free radical inhibition, chelation of ferrous iron, high reducing power, and total antioxidant capacities. Therefore, it can be utilized as a natural antioxidant and applied for dealing with illnesses linked to oxidative stress.

REFERENCES:

1. Zhong Y, Mab C and Shahidi F. Antioxidant and antiviral activities of lipophilic epigallocatechin gallate (EGCG) derivatives. Journal of Functional Foods. 2012; 4: 87–93. https://doi.10.1016/j.jff.2011.08.003.

2. Manikandan R and Vijaya A. A Review on Antioxidant activity of Psidium guajava. Research Journal of Pharmacy and Technology. 2015; 8(3): 339-342. DOI: 10.5958/0974-360X.2015.00056.6

3. Ashfaq M H, Siddique A and Shahid S. Antioxidant Activity of Cinnamon zeylanicum: (A Review). Asian Journal of Pharmaceutical Research. 2021; 11(2): 106-116. DOI: 10. 52711 /2231-5691.2021.00021

4. Rakhi Menon. Antioxidants and their Therapeutic Potential- A Review. Research Journal of Pharmacy and Technology. 2013; 6(12): 1426-1429.

5. Potbhare M and Khobragade D. In Vitro Evaluation of Antioxidant Potential of Ayurvedic Preparations Lauha Bhasma and Mandura Bhasma. Asian Journal of Pharmaceutical Research. 2017; 7(2): 63-66. DOI: 10.5958/2231-.95691.2017.00011

6. Augusti Boligon AA, Machado MM and Athayde ML. Technical Evaluation of Antioxidant Activity. Journal of Medicinal Chemistry. 2014: 4: 517-522. https://doi.10.4172/2161-0444.1000188.

7. Carlson SE, Linder HP and Donoghue MJ. The historical biogeography of Scabiosa (Dipsacaceae): Implications for Old World plant disjunctions. Journal of Biogeography. 2012; 39: 1086–1100. https://doi.org/10.1111/j.1365-2699.2011.02669.x.

8. Pinto D, Rahmouni N, Beghidja N and Silva A. Scabiosa Genus: A Rich Source of Bioactive Metabolites. Medicines. 2018: 5: 110. https://doi.10.3390/medicines5040110.

9. Rigat M, Bonet MÀ, Garcia S, Garnatje T and Vallès J. Studies on pharmaceutical ethnobotany in the high river Ter valley (Pyrenees, Catalonia, Iberian Peninsula). Journal of Ethnopharmacology. 2007: 113: 267–277.\

10. Bonet MÀ and Vallès J. Ethnobotany of Montseny biosphere reserve (Catalonia, Iberian Peninsula): Plants used in veterinar;y medicine. Journal of Ethnopharmacology. 2007: 110: 130–147. https://doi.10.1016/j.jep.2006.09.016.

11. Bussmann RW and Glenn A. Medicinal plants used in Northern Peru for reproductive problems and female health. Journal of Ethnobiology and Ethnomedicine. 2010; 6: 30.

12. Karbab A, Mokhnache K, Arrar L, and Charef N. Total phenolic contents and antioxidant capacity of aqueous extract from Pituranthos scoparius (coss. and dur.) Growingin Algeria. Journal of Drug Delivery and Therapeutics. 2020b: 10(3): 125-127. https://doi.org/10.22270/jddt.v10i3.4096.

13. Karbab A, Charef N, Zarga MHA, Qadri MI and Mubarak MS. Ethnomedicinal documentation and anti-inflammatory effects of n-butanol extract and of four compounds isolated from the stems of Pituranthos scoparius: An invitro and in vivo investigation. Journal of Ethnopharmacology. 2021: 267: 113488. https://doi.org/10.1016/j.jep.2020.113488.

14. Amari S, Karbab A, Arrar L and Charef N. Fractionation, phytochemical screening and antioxidant activity of different sub-fractions from leaves and flowers of Erica arborea L. Turkish Journal of Agriculture - Food Science and Technology. 2023: 11(4): 830-837. https://doi.org/10.24925/turjaf.v11i4.830-837.5698.

15. Rao N, Mittal S, Sudhanshu and Menghani E. Assessment of Phytochemical Screening, Antioxidant and Antibacterial Potential of the Methanolic Extract of Ricinus communis l. Asian Journal of Pharmaceutical and Technology. 2013; 3(1): 20-25.

16. Karbab A, Mokhnache K, Arrar L, Baghiani A, Khennouf S and Charef N. Fractionation, phytochemical screening and free radical scavenging capacity of different subfractions from Pituranthos scoparius roots. Journal of Drug Delivery and Therapeutics. (2020a):10(3), 133136.

17. Derouiche S, Azzi M and Hamida A. Phytochemical Analysis and Antioxidant Property of Rhizome extracts aqueous of Phragmites australis in Alloxan Diabetic Rats. Asian Journal of Pharmaceutical and Technology. 2019; 9(4): 249-252. DOI: 10.5958/2231- 5713.2019.00041.2

18. Charef N, Sebti F, Djarmouni M, Boussoualim N, Baghiani A, Khennouf S, Ourari A, damen MAA, Mubarak MS and Peters, DG. Synthesis, characterization, X-ray structures, and biological activity of some metal complexes of the Schiff base2,2'- (((azanediylbis(propane-3,1diyl)) bis(azanylylidene)) bis(methanylylidene)). Polyhedron. 2015: 85: 450–456. https://doi.org/10.1016/j.poly.2014.09.006.

19. Karbab A, Charef N and Arrar L. Phenolic contents, in vitro antioxidant, and in vivo anti-inflammatory studies of aqueous extract from Pituranthos scoparius (Coss. and Dur.) growing in Algeria. Iranian Journal of Pharmacology and Therapeutics. 2019: 17: 1-7.

20. Prieto P et al. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Analytical Biochemistry. 1999: 337-341.269(2).

21. Jain A, Ojha V, Kumar G, Karthik Land Venkata KBR. Phytochemical Composition and Antioxidant Activity of Methanolic Extract of Ficus benjamina (Moraceae) Leaves. Research Journal of Pharmacy and Technology. 2013; 6 (11): 1184-1189.

22. Amutha K and Victor Arokia D. In Vitro Antioxidant Activity of Ethanolic Extract of Barleria cristata L. Leaves. Research Journal of Pharmacognosy and Phytochemistry. 2009; 1(3): 209-212

23. Kaoudoune C, Benchikh F, Abdennour C, Benabdallah H, Souici C, Derafa I, Mamache W and Amira S. Phenolic Content and Antioxidant Activity of Hydroethanolic and Aqueous Extracts of the Inflorescences of Allium sphaerocephalon L. Research Journal of Pharmacy and Technology. 2024; 17(2): 903-909. DOI: 10.52711/0974-360X.2024.00140.

24. Skała E and Agnieszka S. Dipsacus and scabiosa species-the source of specialized metabolites with high biological relevance. Molecules. 2023: 28: 3754. https://doi.10.3390/molecules28093754.

25. Elhawary SS, Eltantawy ME, Sleem AA, Abdallah HM and Mohamed NM. Investigation of Phenolic Content and Biological Activities of Scabiosa atropurpurea L. World Applied Sciences Journal. 2011:15: 311–317.

26. Ji M, Li SJ, Ma CM. Chemical constituents of the inforescence of Scabiosa comosa Fisch and their antioxide and α-glucosidase inhibitory activities. J Inner Mong Univ (Natural Science Edition). 2014: 4: 398–403

27. Garaev EA, Movsumov IS and Isaev MI. Flavonoids and oleanolic acid from Scabiosa caucasica. Chemistry of Natural Compound. 2008: 44: 520–521. https://doi.org/10.1007/s10600-008-9108-x.

28. Lehbili M, Magid AA, Kabouche A, Voutquenne-Nazabadioko L, Morjani H, Harakat D and Kabouche Z.Triterpenoid saponins from Scabiosa stellata collected in North-eastern Algeria. Phytochemistry. 2018: 150: 40–49. https://doi.org/10.1016/j.phytochem.2018.03.005.

29. Hrichi S, Chaabane-Banaoues R, Bayar S, Flamini G, Majdoub YOE, Mangraviti D, Mondello L, Mzoughi RE, Babba H, Mighri Z and Cacciola F. Botanical and genetic identification followed by investigation of chemical composition and biological activities on the Scabiosa atropurpurea L. stem from Tunisian Flora. Molecules. 2020: 25: 5032-5052. https://doi.10.330./mplecules25215032.

30. Zheng O, Koike K, Han LK, Okuda H and Nikaido T. New Biologically Active Triterpenoid Saponins from Scabiosa tschiliensis. Journal of Natural Product. 2004: 67: 604–613. https://doi.10.1021/np0304722.

31. Toumia IB, Sobeh M, Ponassi M, Banelli B, Dameriha A, Wink M, Ghedira LC and Rosano CA. Methanol Extract of Scabiosa atropurpurea enhances doxorubicin cytotoxicity against resistant colorectal cancer cells in vitro. Molecules. 2020: 25: 5265. https://doi.10.3390/molecules25225265.

32. Kılınc H, Masullo M, Lauro G, D’Urso G, Alankus O, Bifulco G and Piacente S. Scabiosa atropurpurea: A rich source of iridoids with α-glucosidase inhibitory activity evaluated by in vitro and in silico studies. Phytochemistry. 2023: 205: 113471. Doi.10.1016/j.phytochem.2022.113471.

33. Hlilac MB, Mosbah H, Zanina N, Ben Nejma A, Ben Jannet H, Aouni M and Selmi B. Characterisation of phenolic antioxidants in Scabiosa arenaria flowers by LC–ESI‐MS/MS and NMR. Journal of Pharmacy and Pharmacology. 2016: 68(7): 932-940.

34. Christopoulou C, Graikou K and Chinou I. Chemosystematic value of chemical constituents from Scabiosa hymettia (Dipsacaceae). Chemistry and Biodiversity. 2008: 5: 318–323. https://doi.10.1002/cbdv.200890029.

35. Mouffouk C, Hambaba L, Haba H, Mouffouk S, Bensouici C, Mouffouk S, Hachemi M and Khadraoui H. Acute toxicity and in vivo anti-inflammatory effects and in vitro antioxidant and anti-arthritic potential of Scabiosa stellata. Oriental Pharmacy and Experimental Medicine. 2018: 18, 335–348. DOI 10.1007/s13596-018-0320-3.

36. Moharram HA and Youssef MM. Methods for Determining the Antioxidant Activity: A Review. Alexandria Journal of Food Science and Technology. 2014: 11, No. 1, pp. 31-42.

37. Badarinath AV, Mallikarjuna Rao K, Madhu Sudhana Chetty C, Ramkanth S and Rajan TVS. Gnanaprakash,review on in vitro antioxidant methods: comparisions, correlations and consideration. International Journal of Pharmtech Research. 2010:2, 1276-1285.

38. Lehbili M, Magid AA, Hubert J, Kabouche A, Voutquenne-Nazabadioko L, Renault JH, Nuzillard JM, Morjani H, Abedini A, Ganglofd SC and Kabouchea Z. Two new bis-iridoids isolated from Scabiosa stellata and their antibacterial, antioxidant, antityrosinase and cytotoxic activities. Fitoterapia. 2017: 125: 41-48. https://doi.org/10.1016/j.fitote.2017.12.018.

39. Fraga CG, Galleano M, Verstraeten SV and Oteiza PI. Basic biochemical mechanisms behind the health benefits of polyphenols. Molecular Aspects of Medicine. 2010: 31. No.6, 435-445. https://doi.org/10.1016/j.mam.2010.09.006.

40. Kurek-Górecka A, Rzepecka-Stojko A, Górecki M, Stojko J, Sosada M and Grażyna Świerczek-Zięba G. Structure and antioxidant activity of polyphenols derived from propolis. Molecules. 2014:19, 78-101. https://doi.10.3390/molecules19010078.

41. Zhong Y, Mab C and Shahidi F. Antioxidant and antiviral activities of lipophilic epigallocatechin gallate (EGCG) derivatives. Journal of Functional Foods. 2012: 4: 87–93. https://doi.10.1016/j.jff.2011.08.003.

42. Ghosh N, Chakraborty T, Mallick S, Mana S, Singha D and Roy B. Synthesis, characterization and study of antioxidant activity of quercetin-magnesium complex. Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy. 2015: 151: 807–813. https://doi.org/10.1016/j.saa.2015.07.050.

43. Yadav AS and Bhatnagar D. Free radical scavenging activity, metal chelation and antioxidant power of some of the Indian spices. BioFactors. 2007: 31: 219–227. https://doi.10.1002/biof.5520310309.

44. Amorati R and Valgimigli L. Modulation of the antioxidant activity of phenols by non-covalent interactions. Organic Biomolecule Chemistry. 2012: 10: 4147–4158. https://doi: 10.1039/c2ob25174d.

45. Kumar CS, Loh WS, Ooi CW, Quah CK and Fun HK. Structural correlation of some heterocyclic chalcone analogues and evaluation of their antioxidant potential. Molecules. 2013: 18: 11996-12011. https://doi.10.3390/molecules181011996.

46. Mbinda W and Musangi C. Antioxidant activity, total phenolic and total flavonoid contents of stem back and root methanolic extracts of Calotropis procera. Journal of Phytopharmacology. 2019; 8(4): 161-166. doi: 10.31254/phyto.2019.8403.

47. Anjali S and Sheetal S. Phytochemical analysis and free radical scavenging potential of herbal and medicinal plant extracts. Journal of Pharmacognosy and Phytochemistry. 2013: 2: 22-29.

48. Packirisamy S, Gunam V, Mahendra J, Rajendran D and Rajagopal P. Preliminary Phytochemical screening and Antioxidant properties of Methanolic root extract of Picrorhiza kurroa. Research Journal of Pharmacy and Technology. 2023; 16(9): 4266-4270. DOI: 10.52711/0974-360X.2023.00698.

49. Boo YC. Can plant phenolic compounds protect the skin from airborne particulate matter?. Antioxidants. 2019: 8: 379. https://doi.10.3390/antiox8090379.

Received on 16.07.2024 Revised on 14.09.2024

Accepted on 30.10.2024 Published on 24.12.2024

Available online from December 27, 2024

Research J. Pharmacy and Technology. 2024;17(12):6070-6075.

DOI: 10.52711/0974-360X.2024.00920