Untargeted Metabolomics Analysis of Crocus cancellatus subsp. damascenus (Herb.) B. Mathew Stigmas and Their Anticarcinogenic Effect on Breast Cancer Cells.

Safranal, crocin, crocetin, and picrocrocin are major known compounds in the stigma extract of Crocus sativus with various medicinal properties. Crocus cancellatus is another Crocus species that grows extensively in Iran's various regions, such as the Kurdistan province. The predominant metabolites and biological properties of C. cancellatus have not yet been investigated. The ingredients of the stigma ethanol extract of C. cancellatus were investigated using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography with tandem mass spectrometry (LC-MS). The ROIMCR approach was performed to analyze the LC-MS full scan data sets. This method searches the MS regions of interest (ROI) data in the m/z domain and analyses the results using the multivariate curve-resolution alternating least squares (MCR-ALS) chemometrics technique for simultaneous resolution of two extracts. Also, the antiproliferative properties of C. cancellatus against MDA-MB-231 and MCF-7 cancer cells were examined by MTT, dual acridine orange/ethidium bromide test, Annexin V-FITC/PI, and zymography. The GC-MS and LC-MS untargeted metabolomics data analysis of the extract indicated the presence of cytotoxic agents including safranal, crocin, picrocrocin, and crocetin in the stigma ethanol extract of C. cancellatus. Biological tests showed that the viability of MDA-MB-231 and MCF-7 cancer cells is decreased following C. cancellatus treatment in a time- and dose-dependent way in both monolayer and 3D cell cultures. The MCF-7 cell spheroids had greater resistance to the cytotoxic activity of the extract in 3D cell culture than the MDA-MB-231 cell spheroids. The morphological changes of the cells treated with C. cancellatus stigmas extract were indicative of apoptosis. Zymography analysis revealed a similar trend of matrix metallopeptidase-2 (MMP-2) and matrix metallopeptidase-9 (MMP-9) activity in the treated cells with C. cancellatus extract in comparison with doxorubicin treatment as a positive control. The findings of this research indicate that the ethanolic extract of C. cancellatus stigmas was a good source of bioactive metabolites with anticancer activity.

Preparation and characterization of potato starch-thymol dispersion and film as potential antioxidant and antibacterial materials.

The antioxidant/antimicrobial capacity and physical properties of potato starch dispersions enriched with polysorbate-thymol micelle were investigated. Results showed that potato starch have radical scavenging and antibacterial activities only in the presence of polysorbate-thymol but with lower level than polysorbate-thymol alone. The decrease in the antioxidant and antibacterial activities may be attributed to the encapsulation of thymol in the starch chain. Polysorbate-thymol caused a decrease in the particle size and viscosity and an increase in the zeta potential of the starch dispersions. Futhermore, polysorbate-thymol leads to a decrease in the tensile strength, rigidity and swelling, and an increase in the flexibility, solubility and water vapour permeability of the starch films. Atomic force microscopy revealed polysorbate-thymol micelles in the film matrix enclosed by polysaccharide crystal. Scanning electron microscope analysis indicated that polysorbate-thymol caused a coarse film microstructure with the distributed crack in the film matrix. Based on the results, the antioxidant and antibacterial activities of the starch-polysorbate-thymol make starch film suitable for food packaging and preservation.