Traditional Middle Eastern spice blends (Baharat): Antimicrobial activity, metabolomic profile, and trace element analysis

Spices and aromatic herbs have traditionally served culinary, preservative, and health purposes (Liu et al., 2017). This study assessed Middle Eastern spice blends (Baharat) for antimicrobial activity,comprisingblackandredpepper,cinnamon,clove,allspice, garlic, mint, black dry lime, turmeric, cumin, and ginger. EtOH:H2O (3:7)was used to extract the spices individually and in four mixtures (blends 1-4). Blends 2 and 4 exhibited good antibacterial activity through the plate microdilution method (Wiegand et al., 2008) against Gram-positive bacteria (MIC = 0.390 and 3.125 mg/mL against Staphylococcus aureus and Listeria monocytogenes, respec- tively) and inhibition against Enterococcus faecalis (MIC = 12.5 and 6.25 mg/mL for blends 2 and 4, respectively). A reduced inhibitory activity was found against Gram-negative bacteria, except blend 4 against Salmonella typhimurium(MIC=6.25mg/mL).A mild bactericidal activity was shown against Gram-positive bacteria (50-200 mg/mL). The MIC values of positive control (gentamycin) ranged 7-28µL/mL. Blends 2 and 4 underwent metabolomic analysis using UHPLC-HR-Orbitrap/ESI-MS. Altogether, 123 compounds were identified, including phenolic and hydroxycinnamic acids, flavonoids, piperamides, and organosulfur compounds (Ali et al., 2021). Blend 4 also contained nine limonoids exclusive to Citrus L. fruits due to black dry lime inclusion. Finally, ICP- QQQ-MS analyzed 12 trace elements in the spices, comparing their concentrations to FAO/WHO limits (FAO/WHO,2023): Al≤49, Cu≤9.9, Fe≤588, Mn≤484, Zn≤17.4 mg/L; As≤38, Cd≤207, Cr≤636, Hg ≤ 10, Ni ≤ 1266, Pb ≤ 1548, Sn ≤ 60 µL/L. References Ali, A., Wu, H., Ponnampalam, E.N., Cottrell, J.J., Dunshea, F.R., Suleria, H.A.R., 2021. Comprehensive profiling of most widely used spices for their phenolic compounds through LC-ESI-QTOF-MS2 and their antioxidant potential. Antioxidants 10(5), 721. FAO/WHO, 2023. Joint FAO/WHO Food Standards Programme Codex Committee on Contaminants in Foods. Working Document for Information and use in discussions related to contaminants and toxins in the GSCTF. Liu, Q., Meng, X., Li, Y., Zhao, C.N., Tang, G.Y., Li, H.B., 2017. Antibacterial and antifungal activities of spices. J. Mol. Sci. 18 (6), 1283. Wiegand, I., Hilpert, K., Hancock, R.E., 2008. Agar and broth dilution methods to determine the minimal inhibitory concentra- tion (MIC) of antimicrobial substances. Nat Protoc. 3(2), 163–175.