Phenolic Extracts of Zizyphus lotus L. (Rhamnaceae) and Ruta chalepensis L. (Rutaceae) as Alternatives to Antibiotics and their Antimicrobial Effects on Clinical Multidrug-Resistant Pathogens

Year 2022, Volume: 81 Issue: 2, 163 - 183, 29.12.2022

Nour El Houda Bekkar , Meddah Boumediene , Yavuz Selim Cakmak , Bahadır Keskin , Pascal Sonnet

Abstract

Objective: The phytochemical composition and the antibacterial and antifungal properties of Zizyphus lotus L. (ZL) leaves and Ruta chalepensis L. (RC) aerial parts harvested from Oran in northwest Algeria were assessed against multidrug-resistant (MDR) clinical pathogens. Materials and Methods: The phenolic compounds identification in the hydromethanolic (MeOH.E) and the aqueous extracts (Aq. E) was done by HPLC-DAD analysis, while the phenolic, flavonoid and tannin contents were determined using quantitative methods. The antibacterial and antifungal activities were also determined. The synergistic effect between both plants was elucidated using the checkerboard dilution test. Results: An important phenolic content was determined with higher concentrations in Z. lotus leaves extracts than R. chalepensis. The HPLC-DAD analysis allowed us to identify benzoic acid as the major phenolic compound in Z. lotus extracts, while catechin, quercetin and epicatechin were the major compounds identified in R. chalepensis. Important antimicrobial activity was observed against all the clinical pathogen strains. The most potent effect was estimated against MDR Salmonella enterica sp. arizonae with 20±0.1mm of growth inhibition zone diameter using RCMeOH.E, while a diameter of 35.03±0.06 mm was measured using ZLMeOH.E. Also, important anti-Candida activity was estimated. No synergistic interaction against the different microbial strains was determined by applying the combinations of both plants' extracts, with a fractional inhibitory concentration index superior to 4 (FICindex >4). Conclusion: Z. lotus and R. chalepensis can be exploited in the medical field as a potential source of antimicrobial components.

Keywords

Antibiotic alternatives, antimicrobial activity, multidrug-resistant pathogens, phenolic compounds, R. chalepensis, Z. lotus

References

• 1. Karczewska E, Wojtas I, Sito E, Trojanowska D, Budak A, zwolins-ka-Wcislo M, Wilk A. Assessment of Co-Existence of Helicobacter pylori and Candida Fungi in Diseases of the upper Gastrointestinal Tract. J Physiol Pharmacol 2009; 60(6): 33-9. google scholar
• 2. Barbut F, Joly F. The intestinal microbiota: balance and dysbiosis. Pierre and Marie Curie University, Paris-France. 2017. google scholar
• 3. Guo S, Duan JA, Li Y, Wang R, Yan H, Qian D, et al. Comparison of the bioactive components in two seeds of Ziziphus species by different analytical approaches combined with chemometrics. Front Pharmacol 2017; 8: 609. google scholar
• 4. Zhang J, Wu Y, Chen M, Yu B, Wang D, Liu JG. C-glucosyl flavones from Ziziphus jujuba var. spinosa. Chem Nat Compd 2015; 51: 247-51. google scholar
• 5. Hamood A, Hossain A, Humaid MT. Comparative evaluation of total phenols, flavonoids content and antioxidant potential of leaf and fruit extracts of Omani Ziziphus jujuba L. Nat Sci Eng 2015; 18: 78-83. google scholar
• 6. Wu Y, Zhang J, Wang D, Liu JG, Hu Y. Triterpenoid saponins from Ziziphus jujuba var. spinosa. Chem Nat Compd 2013; 49: 677-81. google scholar
• 7. Kang KB, Jang DS, Kim JW, Sung SH. UHPLC-ESI-qTOF-MS analysis of cyclopeptide alkaloids in the seeds of Ziziphus jujuba var. spinosa. MSL 2016; 7: 45-9. google scholar
• 8. Li M, Wang Y, Tsoi B, Jin XJ, He RR, Yao XJ. Indoleacetic acid derivatives from the seeds of Ziziphus jujuba var. spinosa. J Fitote 2014; 99: 48-55. google scholar
• 9. Zhao J, Li SP, Yang FQ, Li P, Wang YT. Simultaneous determination of saponins and fatty acids in Ziziphus jujuba (Suanzaoren) by high performance liquid chromatography- evaporative light scattering detection and pressurized liquid extraction. J Chromatogr A 2006; 11: 188-94. google scholar
• 10. Xie J, Zhang Y, Wang L, Qi W, Zhang M. Composition of fatty oils from Semen Ziziphus spinosae and its cardiotonic effect on isolated toad hearts. Nat Prod Res 2012; 26: 479-83. google scholar
• 11. Zhang Y, Qiao L, Song M, Wang L, Xie J, Feng H. HPLC-ESI-MS/MS analysis of the water-soluble extract from Ziziphi spinosae semen and its ameliorating effect of learning and memory performance in mice. Pharmacogn Mag 2014; 10: 509-16. google scholar
• 12. Chen J, Liu X, Li Z, Qi A, Yao P, Zhou Z, et al. A review of dietary Ziziphus jujuba fruit (Jujube): Developing health food supplements for brain protection. Evid Based Complement Alternat Med 2017; 24: 1-10. google scholar
• 13. Benammar C, Baghdad C, Belarbi M, Subramaniam S, Hichami A, Khan NA. Antidiabetic and antioxidant activities of Zizyphus lotus L. aqueous extracts in Wistar rats. J Nutr Food Sci 2014; 8: 1-6. google scholar
• 14. Bnouham M, Mekhfi H, Legssyer A, Ziyyat A. Ethnopharmacology Forum: Medicinal plants used in the treatment of diabetes in Morocco. Int J Diabetes Metab 2002; 10: 33-50. google scholar
• 15. Borgi W, Ghedira K, Chouchane N. Anti-inflammatory and analgesic activities of Zizyphus lotus root barks. J fitote 2007; 78: 16-9. google scholar
• 16. Abdoul-Azize S. Potential Benefits of Jujube (Zizyphus lotus L.) Bioactive Compounds for Nutrition and Health. J Nutr Metab 2016; 2867470. google scholar
• 17. Mrabti HN, Jaradat N, Kachmar MR, Ed-Dra A, Ouahbi A, Cherrah Y, El Abbes Faouzi M. Integrative Herbal Treatments of Diabetes in Beni Mellal Region of Morocco. J Integr Med 2019; 17: 93-9. google scholar
• 18. Hammi KM, Jdey A, Abdelly C, Majdoub H, Ksouri R. Optimization of ultrasound-assisted extraction of antioxidant compounds from Tunisian Zizyphus lotus fruits using response surface methodology. Food Chem 2015; 184: 80-9. google scholar
• 19. Bencheikh N, Bouhrim M, Kharchoufa L, Mohamed Al Kamaly O, Mechchate H, Es-safi I, et al. The Nephroprotective Effect of Zizy-phus lotus L. (Desf.) Fruits in a Gentamicin-Induced Acute Kidney Injury Model in Rats: A Biochemical and Histopathological Investigation. Molecules 2021; 26(16): 4806. google scholar
• 20. El-Hachimi F, Alfaiz C, Bendriss A, Cherrah Y, Alaoui K. Anti-inflammatory activity of Zizyphus lotus seed oil (L.). J Phyto 2016; 1-8. google scholar
• 21. Borgi W, Recio MC, Rios JL, Chouchane N. Anti-inflammatory and analgesic activities of flavonoid and saponin fractions from Zizy-phus lotus (L.) Lam. J SAJB 2008; 74: 320-24. google scholar
• 22. Benammar. C, Hichami A, Yessoufou A, Simonin AM, Belarbi M, Allali H, et al. Zizyphus lotus L. (Desf.) modulates antioxidant activity and human T-cell proliferation. BMC Complement Altern Med 2010; 10: 54. google scholar
• 23. Wahida B, Abderrahman B, Nabil C. Antiulcerogenic Activity of Zi-zyphus lotus (L.) Extracts. J Ethnopharmacol 2007; 112: 228-31. google scholar
• 24. Bakhtaoui FZ, Lakmichi H, Megraud F, Chait A, Gadhi CEA. Gas-tro-Protective, Anti-Helicobacter Pylori and, Antioxidant Properties of Moroccan Zizyphus lotus L. J Appl Pharm Sci 2014; 4: 81-7. google scholar
• 25. Hossain MA. A phytopharmacological review on the Omani medicinal plant: Ziziphus jujube. J King Saud Univ Sci 2018; 31(4): 1352-7. google scholar
• 26. Marmouzi I, Kharbach M, El-Jemli M, Bouyahya A, Cherrah Y, Bouk-louze A, et al. Antidiabetic, Dermatoprotective, Antioxidant and Chemical Functionalities in Zizyphus lotus Leaves and Fruits. Ind Crops Prod 2019; 132: 134-9. google scholar
• 27. Khouchlaa A, Talbaoui A, El-Yahyaoui EIA, Bouyahya A, Ait-Lahsen S, Kahouadji A, Tijane M. Determination of phenolic compounds and evaluation of in vitro litholytic activity on urinary lithiasis of Zizyphus lotus L. extract of Moroccan origin. J Phyto 2017; 1-6. google scholar
• 28. Ghalem M, Merghache S, Belarbi M. Study on the antioxidant activities of root extracts of Zizyphus lotus from the western region of Algeria. Pharmacogn J 2014; 6: 32-42. google scholar
• 29. Ghedira K. Zizyphus lotus (L.) Desf. (Rhamnaceae): jujubier sauvage. J Phyto 2013; 11: 149-53. google scholar
• 30. Borgi W, Chouchane N. Anti-spasmodic effects of Zizyphus lotus ( L .) Desf. extracts on isolated rat duodenum. J Ethnopharmacol 2009; 126: 571-7. google scholar
• 31. Bencheikh N, Bouhrim M, Kharchoufa L, Mohamed Al-Kamaly O, Mechchate H, Essafi I, et al. The Nephroprotective Effect of Zizy-phus lotus L. (Desf.) Fruits in a Gentamicin-Induced Acute Kidney Injury Model in Rats: A Biochemical and Histopathological Investigation. Molecules 2021; 26: 4806. google scholar
• 32. Gonzalez-Trujano ME, Carrera D, Ventura-Martinez A, Cedillo-Portugal E, Navarrete A. Neuropharmacological profile of an ethanol extract Ruta chalepensis L. in mice. J Ethnoparmacol 2006; 106: 129-35. google scholar
• 33. Loizzo MR, Falco T, Bonesi M, Sicari V, Tundis R, Bruno M. Ruta chalepensis L. (Rutaceae) leaf extract: chemical composition, antioxidant and hypoglicaemic activities. Nat Prod Res 2017; 32. google scholar
• 34. Gomez-Flores R. Antimicrobial effect of chalepensin against Streptococcus mutans. J Med Plant Res 2016; 10(36): 631-4. google scholar
• 35. Szewczyk A, Marino A, Molinari J, Ekiert H, Miceli N. Phytochemical Characterization, and Antioxidant and Antimicrobial Properties of Agitated Cultures of Three Rue Species: Ruta chalepensis, Ruta corsica, and Ruta graveolens. Antiox 2022; 11: 592. google scholar
• 36. Khadhri A, Bouali I, Belkhir S, Mokded R, Smiti S, Falé P. et al. In vitro digestion, antioxidant and antiacetylcholinesterase activities of two species of Ruta: Ruta chalepensis and Ruta montana. Pharmaceutic Biol 2016; 55: 101-7. google scholar
• 37. Adsersen A, Gauguin B, Gudiksen L, Jäger AK. Screening of plants used in Danish folk medicine to treat memory dysfunction for acetylcholinesterase inhibitory activity. J Ethnopharmacol 2006; 104: 418-22. google scholar
• 38. Gonzalez-Trujano ME, Urbina-Trejo E, Santos-Valencia F, Villasa-na-Salazar B, Carmona-Aparicio L, Martinez-Vargas D. Pharmacological and toxicological effects of Ruta chalepensis L. on experimentally induced seizures and electroencephalographic spectral power in mice. J Ethnopharmacol 2021; 271. google scholar
• 39. Kloss WE, Wolfshohl JF. Identification of Staphylococcus species with the API STAPH-IDENT system. J Clin Microbiol 1982; 16(3): 509-16. google scholar
• 40. Beutin L, Montenegro MA, Orskov I, Orskov F, Prada J, Zimmermann S, Stephan R. Close association of verotoxin (Shiga-like toxin) production with enterohemolysin production in strains of Escherichia coli. J Clin Microbiol 1989; 27(11): 2559-64. google scholar
• 41. CLSI: Clinical and Laboratory Standards Institute. Performance standards for Antimicrobial Disk Susceptibility Tests. 364 Approved Standard Twelfth Edition. Clinical and Laboratory Standards Institute, 365 Wayne, PA: 2015. google scholar
• 42. FSM-AC: French Society of Microbiology-Antibiogram Committee. Antibiotics to be tested, concentrations, critical diameters and specific interpretative reading rules. 2013. google scholar
• 43. FMS-AC/EUCAST: French Society of Microbiology-Antibiogram Com-mittee/European Committee on Antimicrobial Susceptibility Testing. Antibiogram Committee of the French Society of Microbiology: 2018. google scholar
• 44. Romani A, Pinelli P, Cantini C, Cimato A, Heimler D. Characterization of Violetto di Toscana, a typical Italian variety of artichoke (Cynara scolymus L.). J Food Chem 2006; 95: 221-5. google scholar
• 45. Chavan UD, Shahidi F, Naczk M. Extraction of condensed tannins from beach pea (Lathyrus maritimus L.) as affected by different solvents. J Food Chem 2001; 75(4): 509-12. google scholar
• 46. Harborne JB. Phytochemical methods: A guide to modern techniques of plant analysis.3rd ed. Chapman & Hall Thomson Sci-ence(UK); 1998. p.203-34. google scholar
• 47. Boizot N, Charpentier JP. Méthode rapide d’évaluation du contenu en composés phénoliques des organes d’un arbre forestier. Cah Tech INRA 2006; 79-82. google scholar
• 48. Samatha T, Shyamsundarachary R, Srinivas P, Swamy NR. Quantification of total phenolic and total flavonoid contents in extracts of Orox-ylum indicum L. Kurz. Asian J Pharmaceutic Clin Res 2012: 5(4): 177-9. google scholar
• 49. Ba K, Tine E, Destain J, Cisse N, Thonart P. Comparative study of phenolic compounds, antioxidant potency of different varieties of Senegalese sorghum and amylolytic enzymes of their malt. Bio-technol Agro Soc Environ 2010; 14(1): 131-9. google scholar
• 50. Sanogo Y, Guessennd NK, Tra-Bi HF, Kouadio NJ, Konan FK, Bamba M, Danho N, Bakayoko A, Ya K, Dosso M. Evaluation in vitro de l’activité des écorces de tige de Anogeissus leiocarpus (DC) Guill. et Perr. (Combretaceae) sur des bactéries responsables de maladies courantes en Afrique et criblage phytochimique. Int J Biol Chem Sci 2016; 10(3): 1139-52. google scholar
• 51. Caponio F, Alloggio V, Gomes T. Phenolic compounds of virgin olive oil: Influence of paste preparation techniques. Food Chem 1999; 64(2): 203-9. google scholar
• 52. Ponce AG, Fritz R, Del-Valle CE, Roura SI. Antimicrobial activity of essential oils on the native microflora of organic Swiss chard. 2003; 36(7): 679-84. google scholar
• 53. Chandrasekaran M, Venkatesalu V. Antibacterial and antifungal activity of Syzygium jambolanum seeds. J Ethnopharmacol 2004; 91(1): 105-8. google scholar
• 54. Orhan G, Bayram A, Zer Y, Balci I. Synergy Tests by E Test and Checkerboard Methods of Antimicrobial Combinations against Brucella melitensis. J Clin Microbiol 2005; 43(1): 140- 3. google scholar
• 55. Mulyaningsih S, Sporer F, Zimmermann S, Reichling J, Wink M. Synergistic properties of the terpenoids aromadendrene and 1,8-cineole from the essential oil of Eucalyptus globulus against antibiotic-susceptible and antibiotic-resistant pathogens. Phytomed 2010; 17: 1061-6. google scholar
• 56. Cheurfa M, Allem R, Zabel K, Aichouni W, Medjkane M. Study of the effects of extracts from the roots of Glycyrrhiza glabra L. and Zizy-phus lotus L. on some human pathogenic bacteria. J Phyto 2017. https://doi.org/10.1007/s10298-017-1116-1. google scholar
• 57. Djemai-Zoughlache S, Yahia M, Hambaba L, Abdeddaim M, Aber-kan MC, Ayachi A. Study of the biological activity of extracts from the fruit of Zizyphus lotus L. TJMPNP 2009; 2: 10- 23. google scholar
• 58. Mansour El-Said S. Studies on Ruta chalepensis, an ancient medicinal herb still used in traditonal medicine. J Ethnopharmacol 1990; 28: 305-12. google scholar
• 59. Loizzo MR, Falco T, Bonesi M, Sicari V, Tundis R, Bruno M. Ruta chalepensis L. (Rutaceae) leaf extract: chemical composition, antioxidant and hypoglicaemic activities. Nat Prod Res 2017; 32. google scholar
• 60. Attou A. Contribution to the phytochemical study and biological activities of the extracts of the plant Ruta chalepensis (Fidjel) from the region of Ain Témouchent. Abou Bekr Belkaid-Tlemcen University. 2011. google scholar
• 61. WHO: World Health Organization. WHO Guidelines on Good Agricultural Practices and Good Harvesting Practices (BPARs) for medicinal plants. 2003. google scholar
• 62. Zouari N, Ayadi I, Fakhfakh , Rebai A, Zouari S. Variation of chemical composition of essential oils in wild populations of Thymus algeriensis Boiss. and Reut., a North African endemic Species. LHD 2012; 11: 28-39. google scholar
• 63. Pinto E, Pina-Vaz C, Salgueiro L, Gonçalves MJ, Costa-De-Olivei-ra S, Cavaleiro C, Palmeira , Rodrigues A, Martinez-De-Oliveira J. Antifungal activity of the essential oil of Thymus pulegioides on Candida, Aspergillus and dermatophyte species. J Med Microbiol 2006;55: 1367-73. google scholar
• 64. Lucchesi ME. Microwave-Assisted Solvent-less Extraction: Design and Application to Essential Oil Extraction. Faculty of Science and Technology, University of Reunion. 2005. google scholar
• 65. Khadri A, Bouali I, Belkhir S, Mokded R, Smiti S, Falé P, Eduarda M, Araujo M, Luisa M, Serralheiro M. In vitro digestion, antioxidant and antiacetylcholinesterase activities of two species of Ruta: Ruta chalepensis and Ruta montana. Pharmaceutic Biol 2016; 55: 101-7. google scholar
• 66. Alotaibi SM, Saleem MS, Al-humaidi JG. Phytochemical contents and biological evaluation of Ruta chalepensis L. growing in Saudi Arabia. SPJ 2018; 26(4): 504-8. google scholar
• 67. Borgi W, Bouraoui A, Chouchane N. Antiulcerogenic activity of Zi-zyphus lotus (L.) extracts. J Ethnopharmacol 2007; 12: 228-31. google scholar
• 68. Chetibi C, Diab S. Study of the biological activity in vitro and in vivo of the methanolic and aqueous extracts of the bark of the roots of Zizyphus lotus L. Frères Mentouri University, Constantine. 2016. google scholar
• 69. Chelli-Chentouf N, Tir-Touil Meddah A, Belgharbi A, Benfreha-Tem-mouri H, Meddah B. Phytochemical and antimicrobial screening of fruits and leaves of Zizyphus lotus L. collected in North West of Algeria. Maghr. J Pure & Appl Sci 2018; 4(1): 50-8. google scholar
• 70. Saiah H, Allem R, El-Kebir FZ. Antioxidant and antibacterial activities of six Algerian medicinal plants. Int J Pharm Pharm Sci 2016; 8(1): 367-74. google scholar
• 71. Belkadi N, Hadj-Ali I. Morphometric study and germination test of jujube seeds (Zizyphus lotus) from southern Algeria. Extraction and dosage of 3 classes of flavonoids and estimation of the effect of fruit powder against Tribolium castaneum Herbst (Coleoptera: Ten-ebrionidae). Mouloud Mammeri University of Tizi-Ouzou, Master’s thesis in Biology. 2016. google scholar
• 72. El-Maaiden E, El-Kharrassi Y, Moustaid K, Essamadi AK, Nasser B. Comparative study of phytochemical profile between Ziziphus spina christi and Ziziphus lotus from Morocco. JFMC 2018. google scholar
• 73. Bensal VK, Goel K. Gastroprotective effect of Acacia nilotica young seed less pod extract: Role of polyphenolic constituents. Asian Pac J Trop Med 2012; 5: 523-8. google scholar
• 74. Ghazghazi H, Aouadhi C, Riahi L, Maaroui A, Hasnaoui B. “Fatty acids composition of Tunisian Ziziphus lotus L. (Desf.) fruits and variation in biological activities between leaf and fruit extracts”. Nat Prod Res 2014; 28(14): 1106-10. google scholar
• 75. Borgi W, Recio MC, Rios JL, Chouchane N. Anti-inflammatory and analgesic activities of flavonoid and saponin fractions from Zizy-phus lotus (L.) Lam. SAJB 2008; 74(2): 320-4. google scholar
• 76. Gonzalez-Trujano ME, Carrera D, Ventura-Martinez A, Cedillo-Portugal E, Navarrete A. Neuropharmacological profile of an ethanol extract Ruta chalepensis L. in mice. J Ethnoparmacol 2006; 106: 129-35. google scholar
• 77. Esameldin E, Joslyn T, Annemarie V. Screening of medicinal plants used in South African traditional medicine for genotoxic effects. Toxicol Lett 2003; 143(2): 195-207. google scholar
• 78. Benammara CH, Benkada D, Benammar F. GC/MS analysis of the essential oil of Ruta chalepensis (Fidjel) from Tlemcen. 2nd Symposium on Aromatic and Medicinal Plants, 2006; Marrakech. google scholar
• 79. Bouajaj S, Romane A, Benyamna A, Amri I, Hanana M, Hamrouni L, Romdhane M. Essential oil composition, phytotoxic and antifungal activities of Ruta chalepensis L. leaves from High Atlas Mountains (Morocco). Nat Prod Res 2014; 28(21): 1910-4. google scholar
• 80. Neffati N, Aloui Z, Karoui H, Guizani I, Boussaid M, Zaouali Y. Phytochemical composition and antioxidant activity of medicinal plants collected from the Tunisian flora. Nat Prod Res 2017; 31(13): 1583-8. google scholar
• 81. Abdoul-Azize S, Bendahmane M, Hichami A, Dramane G, Simonin AM, Benammar C, Sadou H, Akpona S, El-Boustani ES, Khan NA. Effects of Zizyphus lotus L. (Desf.) polyphenols on Jurkat cell signaling and proliferation. Int Immunopharmacol 2013; 15: 364-71. google scholar
• 82. Rsaissi N, El-Kamili Bencharki B, Hillali L, Bouhache M. Antimicrobial activity of fruits extracts of the wild jujube "Ziziphus Lotus (L.) Desf. IJSER 2013; 4(9): 1521-8. google scholar
• 83. Saiah H, Allem R, El-Kebir FZ. Antioxidant and antibacterial activities of six Algerian medicinal plants. Int J Pharm Pharm Sci 2016; 8(1): 367-74. google scholar
• 84. N’guessan AHO, Deliko CED, Bekro JAM, Beko YY. Contents of phenolic compounds of 10 medicinal plants used in the traditional therapy ofarterial hypertension, an emerging pathology in Cote d’Ivoire. 2011; 6: 55-61. google scholar
• 85. Pawlowska AM, Camangi F, Bader A, Braca A. Flavonoids of Zizy-phus jujuba L. and Zizyphus spina-christi (L.) Willd (Rhamnaceae) fruits. Food Chem 2009; 112(4): 858-62. google scholar
• 86. Khadri A, Bouali I, Belkhir S, Mokded R, Smiti S, Fale P, Eduarda M, Araujo M, Luisa M, Serralheiro M. In vitro digestion, antioxidant and antiacetylcholinesterase activities of two species of Ruta: Ruta chalepensis and Ruta montana. Pharmaceutic Biol 2016; 55: 101-7. google scholar
• 87. Loizzo MR, Falco T, Bonesi M, Sicari V, Tundis R, Bruno M. Ruta chalep-ensis L. (Rutaceae) leaf extract: chemical composition, antioxidant and hypoglicaemic activities. Nat Prod Res 2017; 32(5): 521-8. google scholar
• 88. Fakhfakh N, Zouari S, Zouari M, Loussayef C, Zouari N. Chemical composition of volatile compounds and antioxidant activities of essential oil, aqueous and ethanol extracts of wild Tunisian Ruta chalepensis L. (Rutaceae). J Med Plants Res 2012; 6: 593-600. google scholar
• 89. Kacem M, Kacem I, Simon G, Mansourd AB, Chaabouni S, Elfeki A, Bouaziz M. Phytochemicals and biological activities of Ruta chalep-ensis L. growing in Tunisia. Food Biosci 2015; 12: 73-83. google scholar
• 90. Kacem M, Simo G, Leschiera R, Misery L, ElFeki A, Lebonvallet N. Antioxidant and antiinflammatory effects of Ruta chalepensis L. extracts on LPS-stimulated RAW 264.7 cells. In Vitro Cell Dev Biol Anim 2015; 51: 128-41. google scholar
• 91. Gali L, Bedjou F. Antioxidant and anticholinesterase effects of the ethanol extract, ethanol extract fractions and total alkaloids from the cultivated Ruta chalepensis. SAJB 2019; 120: 163-9. google scholar
• 92. Zoughlache S. Study of the biological activity of the fruit extracts of Zizyphus lotus L. El-Hadj Lakhdar University, Batna. 2009. google scholar
• 93. Tlili H, Hanen N, Ben-Arfa A, Neffati M, Boubakri A, Buonocore D, Dossena M, Verri M, Doria D. Biochemical properties and in vitro biological activities of extracts from seven folk medicinal 2 plants growing wild in southern Tunisia. 2019. google scholar
• 94. Wagner H, Bladt S. Plant drug analysis, a thin layer chromatography atlas. 2nd ed. Berlin: Springer-Verlag; 1996. google scholar
• 95. Markham KR. Techniques of flavonoids identification. Ed. London. Academic Press; 1982. google scholar
• 96. Fasla B. Evaluation of the antimitotic and genotoxic potential of medicinal plants and phytochemical analysis. Es-Senia University, Oran. 2009. google scholar
• 97. Bekkar NEH, Meddah B, Cakmak YS, Keskin B. Phenolic Composition, Antioxidant And Antimicrobial Activities of Zizyphus lotus L. and Ruta chalepensis L. Growing in Mascara (Western Algeria). JMBFS 2021; 10(5): e3004-e3004. google scholar
• 98. Cacciola A, D’Angelo V, Raimondo FM, Germano MP, Braca A, De Leo M. Ziziphus lotus (L.) Lam. as a Source of Health Promoting Products: Metabolomic Profile, Antioxidant and Tyrosinase Inhibitory Activities. Chem & Biodiver 2022; e202200237. google scholar
• 99. Marmouzi I, Kharbach M, El-Jemli M, Bouyahya A, Cherrah Y, Bouklouze A, Heyden YV, Faouzi ME. Antidiabetic, dermatopro-tective, antioxidant and chemical functionalities in Zizyphus lotus leaves and fruits. Industrial Crops & Products 2019; 132: 134-9. google scholar
• 100. Dracea O, Larion C, Chifiriuc MC, Raut I, Limban C, Nitulesc GM, Badiceanu CD, Israil AM. New thioureides of 2-(4-methylphe-noxymethyl) benzoic acid with antimicrobial activity. Roum Arch Microbiol Immunol 2008; 3(4): 92-7. google scholar
• 101. Wei Q, Wang X, Cheng JH, Zeng G, Sun DW. Synthesis and antimicrobial activities of novel sorbic and benzoic acid amide derivatives. Food Chem 2018; 268: 220-32. google scholar
• 102. Mkadmini hammi K, Jdey A, Abdelly C, Majdou H, Ksouri R. Optimization of ultrasound-assisted extraction of antioxidant compounds from Tunisian Zizyphus lotus fruits using response surface methodology. Food Chem 2015; 184: 80-9. google scholar
• 103. Tukue M, Karismala MB. Phytochemical screening and antibacterial activity of two common terrestrial medicinal plants Ruta chalep-ensis & Rumex nervosus. Carib J Sci Tech 2014; 2: 634-41. google scholar
• 104. Hamza K, Meziani A. Study of the biological activity of the aqueous extract of Zizyphus lotus leaves. Freres-Mentouri University, Constantine. 2015. google scholar
• 105. Chelli Chentouf N, Tir-Touil Meddah A, Belgharbi A, Benfreha Tem-mouri H, Meddah B. Phytochemical and antimicrobial screening of fruits and leaves of Zizyphus lotus L. collected in North West of Algeria. Maghr J Pure & Appl Sci 2018; 4(1): 50-8. google scholar
• 106. Elaloui M, Ennajah A, Ghazghazi H, Ben-Youssef I, Ben-Othman N, Hajlaoui MR, Khouja A, Laamouri A. Quantification of total phenols, flavonoides and tannins from Ziziphus jujuba (mill.) and Ziziphus lotus (l.) (Desf). Leaf extracts and their effects on antioxidant and antibacterial activities. Int J Sec Metabolite 2017; 4(1): 18-26. google scholar
• 107. Lahmer N, Messai S. Phytochemical and biological study of aqueous and methanolic extracts of Zizyphus lotus root bark (L). Freres-Mentouri University, Constantine. 2017. google scholar
• 108. Garcia-Lafuente A, Guillamon E, Villares A, Mauricio A, Jose R, Martinez A. Flavonoids as anti-inflammatory agents: implications in cancer and cardiovascular disease. Inflamm Res 2009; 58: 537. google scholar
• 109. Amri I, Hamrouni L, Hanana M, Gargouri S, Fezzani T, Jamoussi B. Chemical composition, physicochemical properties, antifungal and herbicidal activities of Pinus halepensis Miller essential oils. Biol Agric Hortic 2013; 29: 91-106. google scholar
• 110. Bukar AM, Kyari MZ, Gwaski M, Gudusu FS, Kuburi PA, Abadam YI. Evaluation of phytochemical and potential antibacterial activity of Ziziphus spina-christi L. against some medically important pathogenic bacteria obtained from University of Maiduguri Teaching Hospital, Maiduguri, Borno State-Nigeria. J Pharmacogn Phyto-chem 2015; 3: 98-101. google scholar
• 111. Naili MB, Alghazeer RO, Saleh NA, Al-Najjar AY. Evaluation of antibacterial and antioxidant activities of Artemisia campestris (Atrace-ae) and Ziziphus lotus (Rhamnaceae). Arab J Chem 2010; 3: 79-84. google scholar
• 112. Zazouli S, Chigr M, Ramos PA, Rosa D, Castro MM, Jouaiti A, et al. Chemical profile of lipophilic fractions of different parts of Zizy-phus lotus L. by GC-MS and evaluation of their antiproliferative and antibacterial activities. Molecules 2022; 27(2): 483. google scholar
• 113. Hammi KM, Jdey A, Abdelly C, Majdou H, Ksouri R. “Optimization of ultrasound-assisted extraction of antioxidant compounds from Tunisian Zizyphus lotus fruits using response surface methodology”. Food Chem 2015; 184: 80-9. google scholar
• 114. Ben-Bnina E, Hammami S, Daamii Remadi M, Ben-Jannet H, Mighri Z. Chemical composition and antimicrobial effects of Tunisian Ruta chalepensis L. essential oils. J Soc Chim Tunisie 2010; 12: 1-9. google scholar
• 115. Haddouchi F, Chaouche TM, Zaouali Y, Ksouri R. Chemical composition and antimicrobial activity of the essential oils from four Ruta species growing in Algeria. Food Chem 2013; 141: 253-8. google scholar
• 116. Amirouche R, Belkola F. In vitro effect of the association of essential oils of Salvia officinalis, Melaleuca alternifolia and two major compounds on bacteria. Faculty of Nature and Life Sciences, University of Mira A, Bejaia. 2013. google scholar
• 117. Ghali A, Rafed S. Phytochemical screening and anti-hemolytic activity of two medicinal plants: Allium ursinum and Allium porrum. University of Bouira. 2019. google scholar