Supplementary data for the article: Aleksić, I., Ristivojević, P., Pavić, A., Radojević, I., Comić, L. R., Vasiljević, B., Opsenica, D., Milojkovic-Opsenica, D., & Šenerović, L. (2018). Anti-quorum sensing activity, toxicity in zebrafish (Danio rerio) embryos and phytochemical characterization of Trapa natans leaf extracts. Journal of Ethnopharmacology, 222, 148–158. https://doi.org/10.1016/j.jep.2018.05.005

Related to published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1111]Related to accepted version:[https://imagine.imgge.bg.ac.rs/handle/123456789/1762]Supplementary material for: [https://doi.org/10.1016/j.jep.2018.05.005

Anti-quorum sensing activity, toxicity in zebrafish (Danio rerio) embryos and phytochemical characterization of Trapa natans leaf extracts

Ethnopharmacological relevance: Trapa natans L. (water chestnut or water caltrop) is a widespread aquatic plant, which has been cultivated for food and traditional medicine since ancient times. Pharmacological studies showed that water chestnut exhibits the wide range of biological activities, such as antimicrobial, antioxidative, analgesic, anti-inflammatory, as well as antiulcer. Aim of the study: Evaluation of anti-virulence potential and toxicity of T. natans methanol (TnM), acetone (TnA) and ethyl acetate (TnEA) leaf extracts. Materials and methods: The anti-quorum sensing activity of Tn extracts was addressed by measuring their effects on biofilm formation, swarming motility and pyocyanin and elastase production in Pseudomonas aeruginosa. Specific P. aeruginosa biosensors were used to identify which of the signaling pathways were affected. The lethal and developmental toxicity of extracts were addressed in vivo using the zebrafish (Danio rerio) model system. The phenolic composition of T. natans leafs extracts was analyzed by a linear ion trap-OrbiTrap hybrid mass spectrometer (LTQ OrbiTrapMS) and UHPLC system configured with a diode array detector (DAD) hyphenated with the triple quadrupole mass spectrometer. Results: Subinhibitory concentrations of Tn leaf extracts (0.2 MIC) inhibited pyocyanin and elastase production up to 50% and 60%, respectively, and reduced swarming zones, comparing to non-treated P. aeruginosa. TnA inhibited biofilm formation by 15%, TnM showed a stimulatory effect on biofilm formation up to 20%, while TnEA showed no effect. The bioactive concentrations of TnM and TnA were not toxic in the zebrafish model system. Twenty-two phenolic compounds were tentatively identified in TnM, where thirteen of them were identified in T. natans for the first time. Tn extracts, as well as their major components, ellagic and ferulic acids, demonstrated the ability to interfere with P. aeruginosa Las and PQS signaling pathways. Conclusions: This study demonstrates anti-virulence potential of Tn leaf extracts against medically important pathogen P. aeruginosa and confirms the ethnopharmacological application of this plant against microbial infections.Related to published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1111]This is the peer reviewed version of the paper: Aleksić, I., Ristivojević, P., Pavić, A., Radojević, I., Comić, L. R., Vasiljević, B., Opsenica, D., Milojkovic-Opsenica, D., & Šenerović, L. (2018). Anti-quorum sensing activity, toxicity in zebrafish (Danio rerio) embryos and phytochemical characterization of Trapa natans leaf extracts. Journal of Ethnopharmacology, 222, 148–158. [https://doi.org/10.1016/j.jep.2018.05.005]Supplementary data: [https://imagine.imgge.bg.ac.rs/handle/123456789/1768

Reversed-phase thin-layer chromatography of some angiotensin converting enzyme (ACE) inhibitors and their active metabolites

The chromatographic behaviour of some ACE inhibitors and their active metabolites was examined under conditions of reversed-phase thin-layer chromatography on RP-18 silica using water–methanol, water–ethanol and water–acetone as binary solvent systems. The relationship between the RM values and the concentration of organic modifier in the mobile phases was linear. It was found that an increase in the content of the organicmodifier in the employed solvent systems led to a decrease of the RM values, i.e., of the retention. Also, the more hydrophobic compounds had a longer retention. Based on regression analysis of the plots, the lipophilicity parameters RM0 and c0 were calculated. The chromatographically obtained lipophilicity parameters were correlated with the calculated log P values

Thin-layer chromatography of several antihypertensive drugs from the group of angiotensin converting enzyme inhibitors

A rapid and simple method for the chromatographic separation of pharmacologically active components contained in some antihypertensive drugs has been developed employing thin-layers of silica gel and polyacrylonitrile sorbent (PANS). The active compounds of Captopril (S)-1-(3-mercapto-2-methyl-1-oxopropyl)-L-proline, Enalapril (S)-1-[N- [1-(ethoxycarbonyl)-3-phenylpropyl]-L-alanyl]-L-proline, Lisinopril (S) -1-[N2-(carboxy-3-phenylpropyl)-L-lysyl]-L-proline, Quinapril [3S-[2[R*(R*)],3R*]], -2- [2-[[1(- etho- xycarbonyl)-3-phenylpropyl]amino]-1-oxopropyl]-1,2,3,4-tetrahydro-3-iso-quinoline-carboxylic acid, Ramipril [2S-[1[R*(R*)],2a3ab,6ab]]-1-[2[[1-(ethoxycarbo- nyl)-3-phenylpropyl]amino]-1-oxopropyl]octahydrocyclopenta[b]-pyrole-2-carboxylic acid and Cilazapril [1S-[1a,9a (R*)]]- 9-[[1-(ethoxycarbonyl)-3-phenylpropyl]amino] octahydro-10-oxo-6H-pyridazino[1,2-a][1,2] diazepine-1-carboxylic acid, were successfully separated by the presented procedures. For their chromatographic separation on silica gel sixteen and on PANS thirteen solvents were used

Mold/aflatoxin contamination of honey bee collected pollen from different Serbian regions

Assessment of microbiological quality of bee collected pollen is very important, because of its use as a supplement in the human diet. In this study, 26 samples collected from different location in Serbia were tested for the presence of mold through mycologial analysis. The presence of aflatoxin B1, one of the most dangerous and the most widespread mycotoxin was also determined. It was established that 10 of the investigated samples were contaminated with some genera or species of mold, but all of the investigated samples were contaminated with aflatoxin B1. Considering that there is no unique and official procedure for mycological analysis of bee collected pollen, these findings suggest the need for their establishment. Mycological analysis should be followed by mycotoxicological analysis since the absence of mold does not confirm the absence of aflatoxin B1 in bee pollen