Evaluation of organic acids, cytotoxicity, oral acute toxicity and diuretic effect of Phaseolus vulgaris crops ethyl acetate extract for cardiovascular health improvement
Abstract
Diuretics play an important role in managing high blood pressure. The aim of this study was to determine the organic acids, toxicity, and diuretic properties of the ethyl acetate fraction of Phaseolus vulgaris (EPV) in Wistar rats. High Performance Liquid Chromatography (HPLC) analysis has been used to detect and quantify the organic acids in the EPV fraction. Cytotoxicity tests of Artemia salina larvae and acute oral toxicity tests are carried out to determine the lethal concentrations and doses of EPV. The diuretic properties of EPV were evaluated in Wistar rats for 24 hours by measuring diuresis parameters and urinary electrolyte concentrations.
Results: The HPLC analysis report showed the presence of various organic acids in EPV, including malic acid (117.79±0.01 mg/g), oxalic acid (52.47±0.00 mg/g), and citric acid (42.85±0.00 mg/g). EPV seems almost non-toxic, as demonstrated by the larvicidal death lethal concentration (LC50) and the estimated 50% lethal dose of 5000 mg/kg body weight. Measuring diuresis parameters reports that EPV may increase urine volume at the test doses. In addition, the report states that EPV affects electrolyte balance by increasing the excretion of Na+, K+, Cl+, and pH. The intensity of the diuretic effect was significantly higher than that of the negative control. These effects are less pronounced than those caused by pharmacological references but are largely similar to those caused by furosemide. A high concentration of oxalic acids, citric acids and ascorbic acids, combined with the proven urine effects of EPV, strengthens the antihypertension, antioxidant and vasorelaxant properties previously reported. These drug properties demonstrate not only the traditional use of EPV but also its use as a database for the development of innovative therapeutic tools to reduce the risk of cardiovascular diseases.
Keywords: Diuretic effect; Organic acids; Phaseolus vulgaris ethyl acetate fraction
Keywords:
Diuretic effect, Organic acid, Phaseolus vulgaris ethy acetate fractionDOI
https://doi.org/10.22270/jddt.v16i5.7765References
1. Pandey M. Clinical pharmacology & therapeutic uses of diuretic agents: A review. J Res Appl Sci Biotechnol. 2022;1(3):11-20. DOI:https://doi.org/10.55544/jrasb.1.3.3
2. Kehrenberg MCA, Bachmann HS. Diuretics: A contemporary pharmacological classification? Naunyn Schmiedebergs Arch Pharmacol. 2022 Jun 1;395(6):619-27. DOI: 10.1007/s00210-022-02228-0 https://doi.org/10.1007/s00210-022-02228-0 PMid:35294605 PMCid:PMC9072265
3. Tamargo J, Segura J, Ruilope LM. Diuretics in the treatment of hypertension. Part 1: Thiazide and thiazide-like diuretics. Expert Opin Pharmacother. 2014 Mar;15(4):527-47. DOI: 10.1517/14656566.2014.879118 https://doi.org/10.1517/14656566.2014.879118 PMid:24444254
4. Morales-Olivas FJ. Diuretics use in the management of hypertension. Hipertens y Riesgo Vasc. 2024 Jul 1;41(3):186-93. DOI: 10.1016/j.hipert.2024.03.004 https://doi.org/10.1016/j.hipert.2024.03.004 PMid:38853071
5. Ferrari F, Zanza C, Tesauro M, Longhitano Y, Planinsic R, Voza A, et al. Clinical Pharmacology of Loop Diuretics in Critical Care. Clin Pharmacokinet [Internet]. 2025 Jul 1 [cited 2025 Oct 8];64(7):987-97. DOI: 10.1007/s40262-025-01524-1 https://doi.org/10.1007/s40262-025-01524-1 PMid:40445471 PMCid:PMC12185637
6. Ellison DH. Clinical pharmacology in diuretic use. Clin J Am Soc Nephrol. 2019 Aug 7;14(8):1248-57. DOI: 10.2215/CJN.09630818 https://doi.org/10.2215/CJN.09630818 PMid:30936153 PMCid:PMC6682831
7. Sica DA. Diuretic‐Related Side Effects: Development and Treatment. J Clin Hypertens. 2007;6(9):532. DOI: 10.1111/j.1524-6175.2004.03789.x https://doi.org/10.1111/j.1524-6175.2004.03789.x PMid:15365284 PMCid:PMC8109680
8. Trullàs JC, Casado J, Morales-Rull JL, Formiga F, Conde-Martel A, Quirós R, et al. Prevalence and outcome of diuretic resistance in heart failure. Intern Emerg Med. 2019 Jun 1;14(4):529-37. DOI: 10.1007/s11739-018-02019-7 https://doi.org/10.1007/s11739-018-02019-7 PMid:30610440
9. Masella C, Viggiano D, Molfino I, Zacchia M, Capolongo G, Anastasio P, et al. Diuretic resistance in cardio-nephrology: role of pharmacokinetics, hypochloremia, and kidney remodeling. Kidney Blood Press Res. 2019 Oct 1;44(5):915-27. DOI: 10.1159/000502648 https://doi.org/10.1159/000502648 PMid:31437845
10. El-Tantawy WH, Temraz A, Mohamed SA. Herbal Extracts Acting as Diuretics: A Comprehensive Review. Egypt J Chem. 2024 Mar 1.DOI: 10.21608/ejchem.2024.254378.8972 https://doi.org/10.21608/ejchem.2024.254378.8972
11. Ouedraogo WRC, Belemnaba L, Nitiema M, Kaboré B, Compaoré S, Koala M, et al. Safety use of Phaseolus vulgaris L. immature pods decoction and cardiovascular effect in rat without hypertension. Int J Pharm Sci Res. 2023;14(10):5050-61. DOI: 10.13040/IJPSR.0975-8232.14(10).5050-61 https://doi.org/10.13040/IJPSR.0975-8232.14(10).5050-61
12. Ouedraogo WRC, Belemnaba L, Nitiema M, Kabore B, Ouedraogo N, Koala M, et al. Antioxidant and Vasorelaxant Properties of Phaseolus vulgaris Linn (Fabaceae) Immature Pods Extract on the Thoracic Aorta of NMRI Mice. Biomed Pharmacol J. 2023;16(1):533-48. DOI : https://dx.doi.org/10.13005/bpj/2635 https://doi.org/10.13005/bpj/2635
13. Salas SP, Russo N M. Analysis of the main ethical conflicts in the 2008 declaration of Helsinki and the proposed changes in the new version. Rev médica Chile. 2014;142(4):475-80. DOI: 10.4067/S0034-98872014000400009 https://doi.org/10.4067/S0034-98872014000400009 PMid:25117038
14. Jospin AF, Boya AA, Abiodoun OP, Fidèle A, Frédéric H. Valeurs nutritives, propriétés antioxydantes et cytotoxicité des cossettes de Dioscorea dumetorum pour la valorisation en alimentation animale et humaine au Bénin. Tropicultura. 2022 Jan 1;40(1). DOI: 10.25518/2295-8010.1963 https://doi.org/10.25518/2295-8010.1963
15. Vanhaecke P, Persoone G, Claus C, Sorgeloos P. Proposal for a short-term toxicity test with Artemia nauplii. Ecotoxicol Environ Saf. 1981;5(3):382-7. DOI: 10.1016/0147-6513(81)90012-9 https://doi.org/10.1016/0147-6513(81)90012-9 PMid:7297475
16. Mousseux M. Test de toxicité sur larves d’ Artemia salina : entretien d’un élevage de balanes. Nouméa : Centre Universitaire de Nouvelle Calédonie; 1995.
17. OCDE. Toxicité orale aiguë - Méthode par classe de toxicité aiguë. OECD; 2001. (Lignes directrices de l’OCDE pour les essais de produits chimiques, Section 4). https://www.oecd-ilibrary.org/environment/essai-n-423-toxicite-orale-aigue-methode-par-classe-de-toxicite-aigue_9789264071018-fr
18. Nitiéma M, Belemnaba L, Ouédraogo S, Ouédraogo N, Ouédraogo S, Guissou IP. Diuretic activity of aqueous decoction extract and ethyl acetate fraction of Lannea microcarpa Engl. and K. Krause (anacardiaceae) trunk barks in wistar rats. World J Pharm Res World J Pharm Res. 2018;7:39-51. DOI:10.20959/wjpr201819-13750
19. Ow CPC, Okazaki N, Iguchi N, Peiris RM, Evans RG, Hood SG, et al. Effects of furosemide, acetazolamide and amiloride on renal cortical and medullary tissue oxygenation in non-anaesthetised healthy sheep. Exp Physiol. 2024 May 1;109(5):766-78. DOI: 10.1113/EP091479 https://doi.org/10.1113/EP091479 PMid:38551893 PMCid:PMC11061632
20. Fitrya, Amriani A, Novita RP, Gabriella R, Lestari SV, Agustina A. The diuretic effect of ethyl acetate fractions of Artocarpus altilis, Artocarpus champeden, and Artocarpus heterophyllus leaves in normotensive Wistar rats. J Ayurveda Integr Med. 2023 Jul 1;14(4):100746. DOI: 10.1016/j.jaim.2023.100746 https://doi.org/10.1016/j.jaim.2023.100746 PMid:37467570 PMCid:PMC10372390
21. Hakim EM, Sivak K V., Kaukhova IE. Evaluation of the diuretic effect of crude ethanol and saponin-rich extracts of Herniaria glabra L. in rats. J Ethnopharmacol. 2021 Jun 12;273:113942. DOI: 10.1016/j.jep.2021.113942 https://doi.org/10.1016/j.jep.2021.113942 PMid:33610714
22. Ngamlai EV, Pradhan RB, Lalbiaknii PC, Ralte V, Lalnunmawia F, Vanlalhluna PC, et al. Diuretic activity evaluation and chemical composition analysis of Hedyotis scandens extract from Mizoram, India, in rat models. J Ethnopharmacol. 2024 Jan 30;319. DOI: 10.1016/j.jep.2023.117079 https://doi.org/10.1016/j.jep.2023.117079 PMid:37659758
23. Ibarrola DA, Arrua W, Gonzalez JE, Soverina Escobar MS, Centurión J, Campuzano Benitez AM, et al. The antihypertensive and diuretic effect of crude root extract and saponins from Solanum sisymbriifolium Lam., in L-NAME-induced hypertension in rats. J Ethnopharmacol. 2022 Nov 15;298:115605. DOI: https://doi.org/10.1016/j.jep.2022.115605 https://doi.org/10.1016/j.jep.2022.115605 PMid:35973627
24. Escudero VJ, Mercadal J, Molina-Andújar A, Piñeiro GJ, Cucchiari D, Jacas A, et al. New Insights Into Diuretic Use to Treat Congestion in the ICU: Beyond Furosemide. Front Nephrol. 2022 Jul 8; 2:879766. DOI: 10.3389/fneph.2022.879766 https://doi.org/10.3389/fneph.2022.879766 PMid:37675009 PMCid:PMC10479653
25. Almajid AN, Cassagnol M. Amiloride. xPharm Compr Pharmacol Ref. 2022 Oct 10;1-6. DOI: 10.1177/1470320320975893 https://doi.org/10.1177/1470320320975893 PMid:33234024 PMCid:PMC7691917
26. Sun Q, Sever P. Amiloride: A review. J Renin-Angiotensin-Aldosterone Syst JRAAS. 2020;21(4). DOI: 10.1177/1470320320975893 https://doi.org/10.1177/1470320320975893 PMid:33234024 PMCid:PMC7691917
27. Salazar-Gómez A, Pablo-Pérez SS, Estévez-Carmona MM, Meléndez-Camargo ME. Diuretic activity of aqueous extract and smoothie preparation of Verbesina crocata in rat. Bangladesh J Pharmacol. 2018 Mar 13;13(3):236-40. DOI: 10.3329/bjp.v13i3.27584 https://doi.org/10.3329/bjp.v13i3.27584
28. Chakraverty R, Samanta K, Mandal P, Karmakar S. Mechanism of action of diuretic and anti-diuretic drugs. How Synth Drugs Work Insights into Mol Pharmacol Class New Pharm. 2023 Jan 1;369-90. DOI: 10.1016/b978-0-323-99855-0.00016-6 https://doi.org/10.1016/B978-0-323-99855-0.00016-6
29. Manvi, Khan MI, Badruddeen, Akhtar J, Ahmad M, Siddiqui Z, et al. Role of Plant Bioactive as Diuretics: General Considerations and Mechanism of Diuresis. Curr Hypertens Rev. 2023 Jun 13;19(2):79-92. DOI: 10.2174/1573402119666230612115220 https://doi.org/10.2174/1573402119666230612115220 PMid:37309769
30. Mariano LNB, Boeing T, da Silva R de CV, da Silva LM, Gasparotto-Júnior A, Cechinel-Filho V, et al. Exotic Medicinal Plants Used in Brazil with Diuretic Properties: A Review. Chem Biodivers. 2022 Jun 1;19(6):e202200258. DOI: 10.1002/cbdv.202200258 https://doi.org/10.1002/cbdv.202200258 PMid:35544364
31. Angappan R, Devanesan AA, Thilagar S. Diuretic effect of chlorogenic acid from traditional medicinal plant Merremia emarginata (Burm. F.) and its by product hippuric acid. Clin Phytoscience 2018 41. 2018 Nov 20;4(1):1-6. DOI:10.1186/s40816-018-0088-5 https://doi.org/10.1186/s40816-018-0088-5
32. Liu Q, Tang GY, Zhao CN, Gan RY, Li H Bin. Antioxidant Activities, Phenolic Profiles, and Organic Acid Contents of Fruit Vinegars. Antioxidants (Basel, Switzerland). 2019 Apr 1;8(4). doi: 10.3390/antiox8040078 https://doi.org/10.3390/antiox8040078 PMid:30934715 PMCid:PMC6523695
33. Us A, Iqbal, Akbar. Furosemide in Acute Kidney Injury - A Vexed Issue. Austin J Nephrol Hypertens Austin J Nephrol Hypertens. 2014;1(1). DOI: 10.1093/ckj/sfx003 https://doi.org/10.1093/ckj/sfx003 PMid:28616210 PMCid:PMC5466115
34. Geldenhuys WJ, Khayat MT, Yun J, Nayeem MA. Drug Delivery and Nanoformulations for the Cardiovascular System. Res Rev Drug Deliv. 2017;1(1). PMID: 28713881; PMCID: PMC5507069.
35. Düsing R. [Diuretics in the treatment of hypertension. Efficacy, safety and tolerability]. Internist (Berl). 2011 Dec;52(12):1484-91. DOI: 10.1007/s00108-011-2915-3 https://doi.org/10.1007/s00108-011-2915-3 PMid:21833757
Published
Abstract Display: 0 
PDF Downloads: 0 PDF Downloads: 0 How to Cite
Issue
Section
Copyright (c) 2026 Windingoudi Rimwagna Christian OUEDRAOGO, Lazare BELEMNABA , Felix Bondo KINI , Fifamin Judith AHOUNOU AÏKPE , Ramona SUHAROSCHI , Floricuta RANGA , Moumouni KOALA , Souleymane COMPAORE , Boukaré KABORE , Mathieu NITIEMA , Sylvin OUEDRAOGO
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0). that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
How to Cite
.