Chemical Composition Related to Antimicrobial Activity of Moroccan Nigella sativa L. Extracts and Isolated Fractions

Abstract

Background. Nigella sativa L. (NS) is an aromatic and medicinal plant commonly used in Mediterranean cuisine. Its grains contain a large amount of fixed oil and have many therapeutic virtues and medicinal properties (antioxidant, antidiabetic, antimicrobial, and anticancer). Aim. The aim of this work is to study the antimicrobial activity of Nigella sativa L. extracts and separated fractions on various pathogenic strains and to correlate that with its chemical composition. Methods. Extracts from Moroccan Nigella sativa seeds were extracted using successive organic solvents, and their hexane and acetone extracts were separated by column chromatography. The chemical composition of extracts, fractions, and essential oil was determined by GC-MS and HPLC-DAD. Extracts and fractions were evaluated for antimicrobial activity through disk diffusion against Gram-positive bacteria (Staphylococcus aureus, Bacillus cereus, and Listeria innocua), Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa), and yeast (Candida pelliculosa) for 1 mg/mL concentration. Bacterial strains were followed to study their behaviors over time in different concentrations. The minimum inhibitory concentration of Nigella sativa essential oil was determined against Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Candida albicans. Results and Conclusion. Although hexane extract was active against both types of bacteria (Gram+ and Gram−), some of its fractions were specifically active against only one type. Fraction (SH4) had the highest activity (15 mm inhibitory diameter). Acetone extract was nonactive but surprisingly resulted in specific active fractions, and the most interesting one was (SA7) that had an inhibitory diameter of 13 mm. This antibacterial effect was related to fatty acids (linoleic and palmitic acids) in (SH4) and 17 pentatriacontene in (SA7). Moreover, the antifungal activity of hexane fractions (10–13 mm) was higher than hexane extract (8 mm), but for acetone, it was the opposite. Acetone extract had a higher activity (18 mm) than its fractions (8–12 mm), except for (SA7) (19 mm). Those inhibitions were attributed to gallic acid, cysteine, and apigenin in acetone extract and cysteine with ascorbic acid in fraction (SA7). Antifungal activity of the essential oil was more pronounced than the antibacterial one. Indeed, determined MICs in the first case were on the microgram scale (MIC = 8 μg/mL, Candida albicans), while in the second case, they were on the milligram scale (MIC = 0.96 mg/mL for Staphylococcus aureus, 0.5 mg/mL for Bacillus cereus, and 0.68 mg/mL for Escherichia coli). This antifungal activity was attributed to three major compounds beta-cymene, alpha-thujene, origanene, and thymoquinone. Results of strains behavior over time at different concentrations of the fractions showed all the curves went through a maximum around 20 hours and had a delay of expression of 5 hours at the start. Taking all results into count, Nigella sativa L. extracts and/or derived principles could form promising antimicrobial agents for therapeutical and industrial uses.