HPLC-HESI-MS/MS Analysis of Phenolic Compounds from Leaf Extracts: An Assessment of Their Cytotoxic, Antioxidant, and Antibacterial Properties.

Dhouha Dallali, Jawhar Fakhfakh, Cédric Paris, Nissaf Aoiadni, Stéphanie Philippot, Arnaud Risler, Mihayl Varbanov, Noureddine Allouche
Author Information
  1. Dhouha Dallali: Laboratory of Organic Chemistry LR17ES08, Natural Substances Team, Faculty of Sciences of Sfax, University of Sfax, Sfax P.O. Box 1171, Tunisia.
  2. Jawhar Fakhfakh: Laboratory of Organic Chemistry LR17ES08, Natural Substances Team, Faculty of Sciences of Sfax, University of Sfax, Sfax P.O. Box 1171, Tunisia.
  3. Cédric Paris: Université de Lorraine, LIBio, F-54000 Nancy, France. ORCID
  4. Nissaf Aoiadni: Laboratory of Animal Eco-Physiology, Faculty of Sciences of Sfax, Sfax P.O. Box 1171, Tunisia.
  5. Stéphanie Philippot: Université de Lorraine, CNRS, L2CM, F-54000 Nancy, France.
  6. Arnaud Risler: Université de Lorraine, CNRS, L2CM, F-54000 Nancy, France.
  7. Mihayl Varbanov: Université de Lorraine, CNRS, L2CM, F-54000 Nancy, France.
  8. Noureddine Allouche: Laboratory of Organic Chemistry LR17ES08, Natural Substances Team, Faculty of Sciences of Sfax, University of Sfax, Sfax P.O. Box 1171, Tunisia. ORCID

Abstract

The current study aimed to investigate the chemical composition, antioxidant, antibacterial, and cytotoxic properties of three extracts (hexane, dichloromethane, and methanol) from The composition of the methanolic extract was elucidated using HPLC-HESI-MS/MS analysis. The antioxidant effect was examined using NO, DPPH, FRAP, and TAC assays. Antimicrobial activity was evaluated by broth microdilution using various bacterial strains such as , , , and . Structural disruptions in Gram-positive bacteria were visualized using scanning electron microscopy (SEM). Cytotoxic effects were evaluated on human MRC-5 in culture according to the MTT assay. The outcomes suggest that methanol extract contained a high amount of phenolic compounds (254.35 0.360 mg GAE/g DE and 211.59 0.939 mg QE/g DE). By applying the HPLC-HESI-MS/MS analysis, 32 compounds were identified, including phenolic acids, flavonoids, lignans, and fatty acids. This extract showed strong antioxidant (IC = 0.043 ± 0.001 mg/mL) and antimicrobial (MIC = 156 µg/mL) activities. The SEM suggests that cells exhibited membrane distortions characterized by surface depressions and alterations in bacterial shape, including dents, when compared to untreated cells. The in vitro cytotoxicity effect on human MRC-5 cells showed no toxicity effects at a concentration of 600 µg/mL. In silico analysis predicted low toxicity for all tested compounds across four different administration routes. This research indicates that this plant could be explored as a powerful source of natural drugs to target pathogens, with applications in the food, pharmaceutical, and medical industries.

Keywords

References

  1. J Cell Mol Med. 2010 Apr;14(4):840-60 [PMID: 19754673]
  2. Antioxidants (Basel). 2022 Sep 19;11(9): [PMID: 36139916]
  3. Planta Med. 2013 May;79(8):616-21 [PMID: 23568627]
  4. Phytochemistry. 2019 May;161:117-129 [PMID: 30825706]
  5. J Agric Food Chem. 2004 Jun 16;52(12):3679-87 [PMID: 15186082]
  6. Phytother Res. 2014 Feb;28(2):224-30 [PMID: 23559516]
  7. Biotechnol Appl Biochem. 2022 Aug;69(4):1757-1764 [PMID: 34490944]
  8. J Pharm Biomed Anal. 2019 Feb 5;164:135-147 [PMID: 30390555]
  9. Lett Appl Microbiol. 2010 Aug;51(2):149-57 [PMID: 20557450]
  10. J Ethnopharmacol. 2012 Jan 6;139(1):149-54 [PMID: 22075456]
  11. Pharmaceuticals (Basel). 2022 May 25;15(6): [PMID: 35745582]
  12. J Chromatogr A. 2006 Jan 13;1102(1-2):184-92 [PMID: 16325827]
  13. J Agric Food Chem. 2014 Oct 8;62(40):9722-35 [PMID: 25266067]
  14. Rapid Commun Mass Spectrom. 2004;18(23):2801-10 [PMID: 15508135]
  15. Malays J Med Sci. 2021 Dec;28(6):64-75 [PMID: 35002491]
  16. Bioresour Technol. 2008 Jul;99(10):4484-94 [PMID: 17935981]
  17. Front Microbiol. 2022 May 04;13:885092 [PMID: 35602020]
  18. BMC Chem. 2022 May 12;16(1):30 [PMID: 35550631]
  19. J Agric Food Chem. 2004 Dec 1;52(24):7251-7 [PMID: 15563203]
  20. Food Chem. 2014 Mar 1;146:289-98 [PMID: 24176345]
  21. Int J Antimicrob Agents. 2011 Jan;37(1):70-4 [PMID: 21075604]
  22. Molecules. 2022 Apr 02;27(7): [PMID: 35408713]
  23. Int J Mol Sci. 2022 Nov 30;23(23): [PMID: 36499380]
  24. Molecules. 2023 May 22;28(10): [PMID: 37241981]
  25. Sichuan Da Xue Xue Bao Yi Xue Ban. 2012 May;43(3):386-90 [PMID: 22812243]
  26. Molecules. 2017 Oct 12;22(10): [PMID: 29023413]
  27. J Toxicol Environ Health B Crit Rev. 2010 Oct;13(7-8):509-26 [PMID: 21170807]
  28. Molecules. 2019 Jan 22;24(3): [PMID: 30678248]
  29. Rapid Commun Mass Spectrom. 2007;21(5):715-29 [PMID: 17279595]
  30. J Med Chem. 2020 Jun 11;63(11):5797-5815 [PMID: 32400157]
  31. FEMS Microbiol Lett. 2022 Aug 8;369(1): [PMID: 35798014]
  32. Microorganisms. 2023 Feb 16;11(2): [PMID: 36838459]
  33. Food Chem. 2014 May 15;151:333-42 [PMID: 24423541]
  34. J Chromatogr Sci. 2013 Mar;51(3):273-85 [PMID: 22977122]
  35. Molecules. 2023 Mar 01;28(5): [PMID: 36903527]
  36. Chem Biodivers. 2020 Dec;17(12):e2000614 [PMID: 33084194]
  37. Food Chem. 2011 Nov 15;129(2):333-344 [PMID: 30634235]
  38. Mar Drugs. 2020 Sep 26;18(10): [PMID: 32993146]
  39. Can Med Assoc J. 1978 Dec 23;119(12):1438-41 [PMID: 737624]
  40. Microorganisms. 2020 Sep 04;8(9): [PMID: 32899776]
  41. Nutrition. 2012 Feb;28(2):197-203 [PMID: 22208556]
  42. J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Feb 15;1139:121992 [PMID: 31991318]
  43. Curr Top Microbiol Immunol. 2017;409:95-120 [PMID: 26667044]
  44. Phytomedicine. 2003 Mar;10(2-3):170-5 [PMID: 12725572]
  45. PhytoKeys. 2022 Mar 15;193:9-42 [PMID: 35586124]
  46. Bioorg Chem. 2020 Mar;96:103535 [PMID: 32000017]
  47. J Mass Spectrom. 2008 Oct;43(10):1421-31 [PMID: 18464298]
  48. Mol Inform. 2011 Mar 14;30(2-3):241-50 [PMID: 27466777]
  49. J Pharm Anal. 2012 Feb;2(1):35-42 [PMID: 29403718]
  50. Biochem Pharmacol. 2015 Dec 1;98(3):371-80 [PMID: 26206193]
  51. J Food Sci. 2011 Aug;76(6):M398-403 [PMID: 22417510]
  52. J Biol Chem. 1981 May 10;256(9):4494-7 [PMID: 7217094]
  53. J Agric Food Chem. 2002 Jun 5;50(12):3579-85 [PMID: 12033832]
  54. Food Chem Toxicol. 2007 Sep;45(9):1731-7 [PMID: 17459553]
  55. World J Microbiol Biotechnol. 2020 Jan 21;36(2):24 [PMID: 31965331]
  56. J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Aug 15;879(24):2459-64 [PMID: 21783434]
  57. Cell Commun Signal. 2023 May 1;21(1):89 [PMID: 37127651]
  58. Molecules. 2013 May 08;18(5):5265-87 [PMID: 23698042]
  59. Molecules. 2020 Aug 09;25(16): [PMID: 32784926]
  60. J Agric Food Chem. 2006 Apr 5;54(7):2745-9 [PMID: 16569070]
  61. Molecules. 2014 Jul 17;19(7):10427-39 [PMID: 25036153]
  62. Chem Biodivers. 2023 Mar;20(3):e202200944 [PMID: 36757004]
  63. Nat Prod Res. 2018 Feb;32(4):481-485 [PMID: 28391725]
  64. J Ethnopharmacol. 2019 Apr 24;234:27-35 [PMID: 30572091]
  65. Bioorg Chem. 2024 Feb;143:107100 [PMID: 38211550]
  66. Food Chem. 2015 Apr 15;173:14-30 [PMID: 25465990]
  67. Molecules. 2020 Mar 24;25(6): [PMID: 32213889]
  68. Int J Biomater. 2022 Jul 22;2022:4427804 [PMID: 35912375]
  69. J Chromatogr A. 2000 Dec 15;902(2):369-79 [PMID: 11192169]
  70. Arch Pharm Res. 2012 Nov;35(11):1919-26 [PMID: 23212633]
  71. Front Microbiol. 2020 Jul 29;11:1829 [PMID: 32849430]

Word Cloud

Created with Highcharts 10.0.0antioxidantusingHPLC-HESI-MS/MS0extractanalysiscompoundscellscompositionmethanoleffectevaluatedbacterialSEMCytotoxiceffectshumanMRC-5phenolicmgDEincludingacidsshowed=antimicrobialµg/mLcytotoxicitytoxicitycurrentstudyaimedinvestigatechemicalantibacterialcytotoxicpropertiesthreeextractshexanedichloromethanemethanolicelucidatedexaminedNODPPHFRAPTACassaysAntimicrobialactivitybrothmicrodilutionvariousstrainsStructuraldisruptionsGram-positivebacteriavisualizedscanningelectronmicroscopycultureaccordingMTTassayoutcomessuggestcontainedhighamount25435360GAE/g21159939QE/gapplying32identifiedflavonoidslignansfattystrongIC043±001mg/mLMIC156activitiessuggestsexhibitedmembranedistortionscharacterizedsurfacedepressionsalterationsshapedentscompareduntreatedvitroconcentration600silicopredictedlowtestedacrossfourdifferentadministrationroutesresearchindicatesplantexploredpowerfulsourcenaturaldrugstargetpathogensapplicationsfoodpharmaceuticalmedicalindustriesAnalysisPhenolicCompoundsLeafExtracts:AssessmentAntioxidantAntibacterialPropertiesCynoglossumtubiflorus

Similar Articles

Cited By