Gas Chromatography‐Mass Spectrometry Analysis and Antiulcerogenic Activity of n‐Butanol Fraction of Balanites aegyptiaca L Delile

Abstract

Introduction The stem bark extract of Balanites aegyptiaca L. (Zygophyllaceae) has been extensively used in traditional medicine as an antiulcer, antiviral and anti-inflammatory agent. Previous scientific studies has established its potency in the treatment of ulcer. The aim of the study was to evaluate the antiulcer activity of the n-butanol fraction of the stem bark extract of Balanites aegyptiaca in Wistar rats and to analyse its components through Gas chromatography-mass spectrometry (GC-MS). Materials and methods Preliminary phytochemical studies were carried out following standard procedures. Oral acute toxicity test to determine the LD50 of the fraction was evaluated in female Wistar rats using OECD 425 guidelines at a limit dose of 2000 mg/kg. The antiulcer activity of the fraction (125, 250 and 500 mg/kg, p.o.) was evaluated in ethanol and indomethacin-induced ulcer models in rats. Rats were divided into five groups (n=5). Group I received distilled water and served as the control while group II received omeprazole 20 mg/kg and served as the standard group. Groups III-V were the fraction treatment groups. At the end of the experiment, mean ulcer indices and percentage ulcer inhibition were assessed. The results were expressed as mean±standard deviation. Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Dunnett's post hoc test. Difference between means were considered statistically significance at p<0.05. The fraction was subjected to further analysis using GC-MS to identify its major components. Results In phytochemical studies, n-butanol fraction of Balanites aegyptiaca showed the presence of alkaloids, tannins, flavonoids, saponins, carbohydrates and glycosides. The acute toxicity test at oral limit test dose of 2000 mg/kg produced neither mortality nor changes related to behavioural, neurologic, autonomic and physical profiles during the short and long term periods. In the ethanol and indomethacin-induced ulcer models, the fraction showed a dose dependent decrease in the mean ulcer inhibition. It was significantly (**p<0.01 and *p<0.05) lower than the control at all dose levels except at 125 mg/kg in the indomethacin induced ulcer model (Figure 1). The GC-MS analysis revealed a total of 17 compounds of which the major ones were Dimethylsulfoxide-D6 (57.53%), Dibutylphthalate (14.19%), n-Hexadecanoic acid (7.62%), 1,2-Benzenedicarboxylic acid, bis(2-methylpropyl) ester (3.69%), Benzaldehyde, 3-hydroxy-4-methoxy (2.17%) and Octadecanoic acid, 2-methylpropyl ester (1.69%). Conclusion The LD50 of n-butanol fraction of Balanites aegyptiaca was found to be greater than 2000 mg/kg and therefore considered safe. The fraction contained alkaloids, tannins, flavonoids, saponins, carbohydrates and glycosides. The data obtained from this study confirmed that the fraction possesses antiulcer activity and contains various bioactive compounds that may be of phytopharmaceutical importance. More studies are on-going to isolate the agent(s) responsible for the activity.