Characterization and antibacterial activity of alkaloids and polyphenols extracts from Haplophyllum tuberculatum (Forssk.) A. Juss.

Abstract

Haplophyllum tuberculatum is a medicinal plant belonging to the Rutaceae family, is renowned for its various therapeutic properties. This study aims to characterize the alkaloids and polyphenol extracts from this plant and assess their antibacterial activity. Herein, the extraction of polyphenols and alkaloids from this plant was performed by the maceration method. Folin Ciacalteu’s method was used to estimate the total phenolic content, and the qualitative characterization of the two extracts was performed by thin-layer chromatography. Whilst, the antibacterial activity of the two extracts was tested with the disk diffusion method on a solid medium and the minimal inhibitory concentration (MIC) of susceptible bacteria was determined using the agar dilution method. Our results indicate respective yields of 8.39% in polyphenols and 0.37% in alkaloid extracts, while the total phenolic content was estimated to be 74.45 mg GAE/g of dry matter in polyphenolic extract. Thin-layer chromatography analysis allowed choosing the system toluene-acetate-ethanol-concentrated ammonia (40:4:8:3, v/v) to separate H. tuberculatum alkaloids, and ethyl acetate-methanol-water (100:13.5:10, v/v/) for the separation of its polyphenols. The same analysis detected traces of quercitin, catechin, and rutin in the polyphenolic extract. Our findings demonstrated good antibacterial activity on Gram-positive strains such as Staphylococcus aureus ATCC 25923 and Bacillus subtilis ATCC 6633, and moderate activity on Pseudomonas aeruginosa ATCC 27953, with MICs ranging from 0.625 to 10 mg/mL for alkaloids and from 5 to 20 mg/mL for polyphenols.

Keywords

• Haplophyllum tuberculatum
• Alkaloids
• Polyphenols
• Maceration
• Antibacterial activity

Supporting Institution

/

Project Number

/

Thanks

/

References

1. Abdelgaleil, S., Saad, M., Ariefta, N., & Shiono, Y. (2020). Antimicrobial and phytotoxic activities of secondary metabolites from Haplophyllum tuberculatum and Chrysanthemum coronarium. South African Journal of Botany, 128, 35 41. https://doi.org/10.1016/j.sajb.2019.10.005
2. Abdelkhalek, A., Salem, M.Z., Hafez, E., Behiry, S.I., & Qari, S.H. (2020). The phytochemical, antifungal, and first report of the antiviral properties of Egyptian Haplophyllum tuberculatum extract. Biology, 9(9), 248. https://doi.org/10.3390/biology9090248
3. Aboshora, W., Lianfu, Z., Dahir, M., Qingran, M., Qingrui, S., Jing, L., Al-Haj, N.Q.M., & Ammar, A. (2014). Effect of extraction method and solvent power on polyphenol and flavonoid levels in Hyphaene thebaica L mart (Arecaceae) (Doum) fruit, and its antioxidant and antibacterial activities. Tropical Journal of Pharmaceutical Research, 13(12), 2057-2063. https://doi.org/10.4314/tjpr.v13i12.16
4. Abou-Zeid, H.M., Bidak, L.M., & Gohar, Y.M. (2014). Phytochemical screening and antimicrobial activities of some wild medicinal plants of the western mediterranean coastal region, Egypt. International Journal of Pharmaceutical Sciences and Research 5(7), 3072-3080. http://doi.org/10.13040/IJPSR.0975-8232.5(10).4445-55
5. Al-Brashdi, A.S., Al-Ariymi, H., Al Hashmi, M., & Khan, S.A. (2016). Evaluation of antioxidant potential, total phenolic content and phytochemical screening of aerial parts of a folkloric medicine, Haplophyllum tuberculatum (Forssk) A. Juss. Juss Journal of Coastal Life Medicine, 4, 315-319. https://doi.org/10.12980/jclm.4.2016J6-4
6. Al-Burtamani, S.K.S., Fatope, M.O., Marwah, R.G., Onifade, A.K., & Al-Saidi, S.H. (2005). Chemical composition, antibacterial and antifungal activities of the essential oil of Haplophyllum tuberculatum from Oman. Journal of Ethnopharmacology, 96(1-2), 107-112. https://doi.org/10.1016/j.jep.2004.08.039
7. Al-Rehaily, A.J., Al-Howiriny, T.A., Ahmad, M.S., Al-Yahya, M.A., El-Feraly, F.S., Hufford, C.D., & McPhail, A.T. (2001). Alkaloids from Haplophyllum tuberculatum. Phytochemistry, 57(4), 597-602. https://doi.org/10.1016/S0031-9422(01)00041-3
8. Al-Saeghi, S.S., Hossain, M.A., & Al-Touby, S.S.J. (2022). Characterization of antioxidant and antibacterial compounds from aerial parts of Haplophyllum tuberculatum. Journal of Bioresources and Bioproducts, 7(1), 52-62. https://doi.org/10.1016/j.jobab.2021.09.004

9. Al-Said, M.S., Tariq, M., Al-Yahya, M.A., Rafatullah, S., Ginnawi, O.T., & Ageel, A.M. (1990). Studies on Ruta chalepensis, an ancient medicinal herb still used in traditional medicine. Journal of Ethnopharmacology, 28(3), 305-312. https://doi.org/10.1016/0378-8741(90)90081-4
10. Al-Shamma, A., Al-Douri, N.A., & Phillipson, J.D. (1979). Alkaloids of Haplophyllum tuberculatum from Iraq. Phytochemistry, 18(8), 1417-1419. https://doi.org/10.1016/0031-9422(79)83044-7
11. Alvarez, A.I., Real, R., Pérez, M., Mendoza, G., Prieto, J.G., & Merino, G. (2010). Modulation of the activity of ABC transporters (P-glycoprotein, MRP2, BCRP) by flavonoids and drug response. Journal of Pharmaceutical Sciences, 99(2), 598 617. https://doi.org/10.1002/jps.21851
12. Al-Yahya, M.A., Al-Rehaily, A.J., Ahmad, M.S., Al-Said, M.S., El-Feraly, F.S., & Hufford, C.D. (1992). New alkaloids from Haplophyllum tuberculatum. Journal of Natural Products, 55(7), 899-903. https://doi.org/10.1021/np50085a008
13. Amhis, W., Benslimane, A., Tiouit, D., & Naim, M. (2001). Tests de sensibilité utiles au traitement antibiotique [Useful sensitivity tests for antibiotic treatment]. Médecine du Maghreb, 91, 22-25. http://www.santetropicale.com/Resume/9105.pdf
14. Balouiri, M., Sadiki, M., & Ibnsouda, S.K. (2016). Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis, 6(2), 71 79. https://doi.org/10.1016/j.jpha.2015.11.005
15. Bansemir, A., Blume, M., Schröder, S., & Lindequist, U. (2006). Screening of cultivated seaweeds for antibacterial activity against fish pathogenic bacteria. Aquaculture, 252(1), 79-84. http://doi.org/10.1016/j.aquaculture.2005.11.051
16. Benchelah, A.C., Bouziane, H., & Maka, M. (2004). Fleurs du Sahara, arbres et arbustes, voyage au coeur de leurs usages avec les Touaregs du Tassili [Flowers of the Sahara, trees and shrubs, journey to the heart of their uses with the Tuaregs of Tassili]. Phytothérapie, 2, 191-197. https://doi.org/10.1007/s10298-004-0052-z
17. Bouzid, W., Yahia, M., Abdeddaim, M., Aberkane, M., & Ayachi, A. (2011). Evaluation of antioxidant and antimicrobial activities of extract of Crataegus monogyna (hawthorn). Lebanese Science Journal. 12, 59 69. https://inis.iaea.org/search/search.aspx?orig_q=RN:44051613
18. Bruneton, J., & Hatton, C.K. (1999). Pharmacognosy, phytochemistry, medicinal plants (2nd ed.). Intercept, Lavoisier publishing.
19. Catier, O., & Roux, D. (2007). Botanique pharmacognosie phytothérapie: Cahiers du préparateur en pharmacie [Botany pharmacognosy phytotherapy: Pharmacy preparer's notebooks] (3rd ed.). Groupe Liaisons. Netherlands.
20. Çolak, F., Savaroğlu, F., & İlhan, S. (2009). Antibacterial and antifungal activities of Arum maculatum L. leaves extracts. Journal of Applied Biological Sciences, 3(3), 13-16. https://dergipark.org.tr/en/pub/jabs/issue/34906/387137.
21. Daglia, M. (2012). Polyphenols as antimicrobial agents. Current Opinion in Biotechnology, 23(2), 174-181. https://doi.org/10.1016/j.copbio.2011.08.007
22. Diallo, A. (2005). Etude des plantes médicinales de Niafunke (Region de Tombouctou): Phytochimie et pharmacologie de Maerua crassifolia Forsk (Capparidaceae) [Study of medicinal plants from Niafunke (Timbuktu region): Phytochemistry and pharmacology of Maerua crassifolia Forsk (Capparidaceae)]. [Doctoral dissertation, University of Bamako]. Bamako University, Mali. https://pesquisa.bvsalud.org/portal/resource/pt/afr-190537?lang=fr
23. Dohou, R., Yamni, K., Tahrouch, S., Hassani, L.I., Badoc, A., & Gmira, N. (2003). Screening phytochimique d'une endémique iberomarocaine, Thymelaea lythroides [Phytochemical screening of an Ibero-Moroccan endemic, Thymelaea lythroides]. Bulletin-Société de Pharmacie de Bordeaux, 142(1/4), 61-78.
24. Falleh, H., Ksouri, R., Chaieb, K., Karray-Bouraoui, N., Trabelsi, N., Boulaaba, M., & Abdelly, C. (2008). Phenolic composition of Cynara cardunculus L. organs, and their biological activities. Comptes Rendus Biologies, 331(5), 372 379. https://doi.org/10.1016/j.crvi.2008.02.008
25. Ferrari, J. (2002). Contribution à la connaissance du métabolisme secondaire des Thymelaeaceae et investigation phytochimique de l'une d'elles:" Gnidia involucrata" Steud. ex A. Rich. [Contribution to the knowledge of the secondary metabolism of Thymelaeaceae and phytochemical investigation of one of them: "Gnidia involucrata" Steud. ex A. Rich.] [Doctoral dissertation, Lausanne University]. Lausanne University, Switzerland] https://patrinum.ch/record/15861/files/These_FerrariJ.pdf
26. Hadjadj, S., Bayoussef, Z., El Hadj-Khelil, A.O., Beggat, H., Bouhafs, Z., Boukaka, Y., Khaldi, I.A., Mimouni, S., Sayah, F., Meriem, T. (2015). Ethnobotanical study and phytochemical screening of six medicinal plants used in traditional medicine in the Northeastern Sahara of Algeria (area of Ouargla). Journal of Medicinal Plants Research, 9(41), 1049-1059. https://doi.org/10.5897/JMPR2015.5932
27. Hamdi, A., Halouani, A., Aouf, I., Viaene, J., Marzouk, B., Kraiem, J., & Vander Heyden, Y. (2021). Cytotoxicity and antiviral activities of Haplophyllum tuberculatum essential oils, pure compounds, and their combinations against Coxsackievirus B3 and B4. Planta Medica, 87(10/11), 827-835. https://doi.org/10.1055/a-1538-5289
28. Hayouni, E.A., Abedrabba, M., Bouix, M., & Hamdi, M. (2007). The effects of solvents and extraction method on the phenolic contents and biological activities in vitro of Tunisian Quercus coccifera L. and Juniperus phoenicea L. fruit extracts. Food Chemistry, 105(3), 1126-1134. https://doi.org/10.1016/j.foodchem.2007.02.010
29. Hoffmann, D. (2003). Medical herbalism: The science and practice of herbal medicine. Simon and Schuster.
30. Iserin, P. (2001). Larousse encyclopédie des plantes médicinales : Identification, préparations, soins [Larousse encyclopedia of medicinal plants: Identification, preparations, care] (2nd ed.). DorlingKindersiey Limited, Londres.
31. Karou, D., Dicko, M.H., Simpore, J., & Traore, A.S. (2005). Antioxidant and antibacterial activities of polyphenols from ethnomedicinal plants of Burkina Faso. African Journal of Biotechnology, 4(8), 823. http://www.academicjournals.org/AJB
32. Lee, K.W., Kim, Y.J., Lee, H.J., & Lee, C.Y. (2003). Cocoa has more phenolic phytochemicals and a higher antioxidant capacity than teas and red wine. Journal of Agricultural and Food Chemistry, 51(25), 7292-7295. https://doi.org/10.1021/jf0344385
33. Miliauskas, G., Venskutonis, P., & Van Beek, T. (2004). Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry, 85(2), 231-237. https://doi.org/10.1016/j.foodchem.2003.05.007
34. Muylaert, A., & Mainil, J. (2013). Résistance bactériennes aux antibiotiques, les mécanismes et leur contagiosité [Bacterial resistance to antibiotics, mechanisms and their contagiousness]. Annales de Médecine vétérinaire, 156, 109 123. https://orbi.uliege.be/bitstream/2268/168957/1/R%C3%A9sistances%20bact%C3%A9riennes%20antiobio.pdf
35. Owen, P.L., & Johns, T. (1999). Xanthine oxidase inhibitory activity of northeastern North American plant remedies used for gout. Journal of Ethnopharmacology, 64(2), 149-160. https://doi.org/10.1016/S0378-8741(98)00119-6
36. Pavić, V., Flačer, D., Jakovljević, M., Molnar, M., & Jokić, S. (2019). Assessment of total phenolic content, in vitro antioxidant and antibacterial activity of Ruta graveolens L. extracts obtained by choline chloride based natural deep eutectic solvents. Plants, 8(3), 69. https://doi.org/10.3390/plants8030069
37. Ponce, A.G., Fritz, R., Del Valle, C., & Roura, S.I. (2003). Antimicrobial activity of essential oils on the native microflora of organic Swiss chard. LWT-Food Science and Technology, 36(7), 679-684. https://doi.org/10.1016/S0023-6438(03)00088-4
38. Raissi, A., Arbabi, M., Roustakhiz, J., & Hosseini, M. (2016). Haplophyllum tuberculatum: An overview. Journal of HerbMed Pharmacology, 5(4), 125 130. http://herbmedpharmacol.com/Article/JHP_1044_20160509005140
39. Salvo, G., Manafzadeh, S., Ghahremaninejad, F., Tojibaev, K., Zeltner, L., & Conti, E. (2011). Phylogeny, morphology, and biogeography of Haplophyllum (Rutaceae), a species‐rich genus of the Irano Turanian floristic region. Taxon, 60(2), 513 527. https://doi.org/10.1002/tax.602018
40. Sarni-Manchado, P., & Cheynier, V. (2006). Les polyphénols en agroalimentaire [Polyphenols in food industry]. Collection Sciences et techniques agro-alimentaires, Tec & Doc Editions. Paris.
41. Shan, B., Cai, Y.-Z., Brooks, J.D., & Corke, H. (2007). The in vitro antibacterial activity of dietary spice and medicinal herb extracts. International Journal of Food Microbiology, 117(1), 112-119. https://doi.org/10.1016/j.ijfoodmicro.2007.03.003
42. Sheriha, G.M., & Abou Amer, K.M. (1984). Lignans of Haplophyllum tuberculatum. Phytochemistry, 23(1), 151-153. https://doi.org/10.1016/0031-9422(84)83096-4
43. Singh, A., Palariya, D., Dhami, A., Prakash, O., Kumar, R., Rawat, D., & Pant, A. (2020). Biological activities and phytochemical analysis of Zanthoxylum armatum DC. leaves and bark extracts collected from Kumaun region, Uttarakhand, India. Journal of Medicinal Herbs and Ethnomedicine, 6, 1-10. https://doi.org/10.25081/jmhe.2020.v6.5754
44. Touati, N., Saidani, K., Boudries, H., Hammiche, H., Ouazene, N., & Bedjou, F. (2018). Antibacterial activity of phenolic compounds of Pulicaria odora, wild plant in northern Algeria. International Food Research Journal, 25(5), 2021 2030. https://www.proquest.com/openview/7e79d500e3a361eb929f1dedf3651ae1/1?pq-origsite=gscholar&cbl=816390
45. Tyihák, E., Móricz, Á., & Ott, P. (2008). Biodetection and determination of biological activity of natural compounds. In Waksmundzka-Hajnos, M., Sherma, J., & Kowalska T. (eds), Thin Layer Chromatography in Phytochemistry, Chromatographic Science Series, Vol 99, (pp. 193-213), CRC Press Taylor & Francis Group. http://doi.org/10.1201/9781420046786.ch9
46. Wong, C.-C., Li, H.-B., Cheng, K.-W., & Chen, F. (2006). A systematic survey of antioxidant activity of 30 Chinese medicinal plants using the ferric reducing antioxidant power assay. Food Chemistry, 97(4), 705-711. https://doi.org/10.1016/j.foodchem.2005.05.049