Comparison of ethanolic and ethyl acetate fractions of Iraqi Medicago sativa for the treatment of urinary tract infection

Ahmed Ibrahim  Al-Yousif; Sheelan Amir  Al-Darwish; Farah  Fawzi; Alaa Ghaith  Ahmed

doi:10.31450/ukrjnd.2(86).2025.01

Ahmed Ibrahim Al-Yousif College of Pharmacy, Ashur University, Baghdad, Iraq https://orcid.org/0000-0003-1074-7661
Sheelan Amir Al-Darwish Al-Rasheed University College, Baghdad, Iraq https://orcid.org/0009-0000-6010-7553
Farah Fawzi College of Pharmacy, Al-Bayan University, Baghdad, Iraq https://orcid.org/0009-0003-4049-1326
Alaa Ghaith Ahmed Dr. Saad Al-Witry Neuroscience Hospital, Baghdad, Iraq https://orcid.org/0000-0003-2708-1632

DOI: https://doi.org/10.31450/ukrjnd.2(86).2025.01

Keywords: urinary tract infection, antibiotic, ethyl acetate fraction, M. sativa extract, ethanolic fraction.

Abstract

Urinary tract infections (UTIs), predominantly caused by uropathogenic Escherichia coli (UPEC), affect over 100 million people annually and are a leading cause of morbidity due to rising antibiotic resistance. Medicago sativa (alfalfa), a medicinal plant rich in phytochemicals, has shown antibacterial potential, yet its efficacy against UPEC in Iraq remains unexplored. This study investigates the antibacterial effects of M. sativa ethanolic and ethyl acetate fractions as potential alternatives to conventional antibiotics for UTI treatment.

Methods. M. sativa was collected in Kirkuk, defatted with hexane, extracted with 85% ethanol, and fractionated into petroleum ether, chloroform, ethyl acetate, and ethanolic fractions. Phytochemical analyses, including Dragendorff’s, Mayer’s, and ррigh-performance liquid chromatography were performed. Urine samples from 85 UTI patients were cultured, yielding 30 UPEC isolates. Antibacterial activity was evaluated using the agar well diffusion method, with minimal inhibitory concentrations (MICs) determined via microplate serial dilution. Antibiotic susceptibility was tested using the Kirby-Bauer method against eight antibiotics. Data was analyzed using SPSS v26 (ANOVA, LSD).

Results. Most participants (56.7%) were under 40 years old, with females more affected. The ethanolic fraction demonstrated superior antibacterial activity, with a mean inhibition zone of 21.96 ± 1.9 mm at 75 mg/ml (p=0.001), compared to 17.32 ± 1.5 mm for the ethyl acetate fraction. High-performance liquid chromatography confirmed bioactive compounds, including gallic acid, quercetin, and myricetin. Meropenem exhibited 100% sensitivity, while cephalothin showed complete resistance.

Conclusions. M. sativa extracts, particularly the ethanolic fraction, exhibit significant antibacterial activity against UPEC, offering a promising alternative to antibiotics. Larger, multicenter studies are needed to validate these findings and explore clinical applications.

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