Saffron Stigma Research Articles

Potential Role of Phytochemical Extract from Saffron in Development of Functional Foods and Protection of Brain-Related Disorders.

The present review is designed to measure the effects of saffron extract in functional foods and its pharmacological properties against various disorders. Saffron is a traditional medicinal plant used as a food additive. The stigma of saffron has bioactive compounds such as safranal, crocin, crocetin, picrocrocin, kaempferol, and flavonoid. These bioactive compounds can be extracted using conventional (maceration, solvent extraction, soxhlet extraction, and vapor or hydrodistillation) and novel techniques (emulsion liquid membrane extraction, ultrasound-assisted extraction, enzyme-associated extraction, pulsed electric field extraction, microwave-assisted extraction, and supercritical fluid extraction). Saffron is used as a functional ingredient, natural colorant, shelf-life enhancer, and fortifying agent in developing different food products. The demand for saffron has been increasing in the pharma industry due to its protection against cardiovascular and Alzheimer disease and its antioxidant, anti-inflammatory, antitumor, and antidepressant properties. Conclusively, the phytochemical compounds of saffron improve the nutrition value of products and protect humans against various disorders.

Saffron essential oil ameliorates CUMS-induced depression-like behavior in mice via the MAPK-CREB1-BDNF signaling pathway

Ethnopharmacological relevanceSaffron, the dried stigma of Crocus sativus L., has a long history of use in the treatment of depression in traditional Chinese medicine and Islamic medicine. The unique aroma of saffron, primarily derived from its volatile oil, has been widely used by folk to mitigate anxiety and depression via sniffing because the aroma of saffron has a pleasant and invigorating effect.Aim of the study: This study aimed to investigate the antidepressant effect and the underlying mechanism of saffron essential oil (SEO) in mice exposed to chronic unpredictable mild stress (CUMS). Materials and methodsIn this study, compounds of SEO were identified using gas chromatography-mass spectrometry analysis, while network pharmacology was used to predict potential active compounds, antidepressant targets, and related signaling pathways of SEO. The CUMS depression model was further used to explore the therapeutic effect and possible mechanism of SEO. During the modeling period, mice were regularly administered fluoxetine (3.6 mg/kg, i.g.) or diluted SEO (2%, 4%, and 6% SEO, inhalation). The antidepressant and neuroprotective effects of SEO were evaluated by behavior tests (the open field test, the sucrose preference test, the tail suspension test, and the forced swimming test), hematoxylin-eosin staining, and Nissl staining. The enzyme-linked immunosorbent assay kits were used to measure dopamine (DA), 5-serotonin (5-HT), brain-derived neurotrophic factor (BDNF), and γ-aminobutyric acid (GABA) levels in serum. The relative abundance of Raf1, MEK1, P-ERK1/2/ERK1/2, P-CREB1/CREB1, BDNF, and P-Trk B/Trk B in the hippocampus was determined using western blot (WB). ResultsAccording to the network pharmacology analysis, seven active SEO compounds mediated 113 targets related to depression treatment, most of which were enriched in the 5-HT synapse, calcium signaling pathway, and cAMP signaling pathway. In vivo experiments indicated that fluoxetine and SEO improved depression-like behaviors in depressed mice. The levels of 5-HT, DA, BDNF, and GABA in serum increased significantly. Histopathological examinations revealed that fluoxetine and SEO ameliorated neuronal damage in the hippocampus. WB analysis showed that the relative expressions of Raf1, MEK1, P-ERK1/2/ERK1/2, P-CREB1/CREB1, BDNF, and P-Trk B/Trk B were significantly higher in the fluoxetine and SEO groups than in the CUMS group. ConclusionOverall, these findings suggest that SEO significantly alleviates the depressive symptoms in CUMS exposed mice and partially restores hippocampal neuronal damage. Meanwhile, the best efficacy was observed in 4% SEO. Furthermore, the antidepressant mechanism of SEO is primarily dependent on the regulation of the MAPK-CREB1-BDNF signaling pathway.

Comparative Metabolomics Analysis of Stigmas and Petals in Chinese Saffron (Crocus sativus) by Widely Targeted Metabolomics.

The dried stigmas of Crocus sativus, commonly known as saffron, are consumed largely worldwide because it is highly valuable in foods and has biological activities beneficial for health. Saffron has important economic and medicinal value, and thus, its planting area and global production are increasing. Petals, which are a by-product of the stigmas, have not been fully utilized at present. We compared the metabolites between the stigmas and petals of C. sativus using a non-targeted metabolomics method. In total, over 800 metabolites were detected and categorized into 35 classes, including alkaloids, flavonoids, amino acids and derivatives, phenols and phenol esters, phenylpropanoids, fatty acyls, steroids and steroid derivatives, vitamins, and other metabolites. The metabolite composition in the petals and stigmas was basically similar. The results of the study showed that the petals contained flavonoids, alkaloids, coumarins, and other medicinal components, as well as amino acids, carbohydrates, vitamins, and other nutritional components. A principal components analysis (PCA) and an orthogonal partial least-squares discriminant analysis (OPLS-DA) were performed to screen the different metabolic components. A total of 339 differential metabolites were identified, with 55 metabolites up-regulated and 284 down-regulated. The up-regulated metabolites, including rutin, delphinidin-3-O-glucoside, isoquercitrin, syringaresinol-di-O-glucoside, dihydrorobinetin, quercetin, and gallocatechin, were detected in the petals. The down-regulated metabolites were mainly glucofrangulin B, acetovanillone, daidzein, guaiazulene, hypaphorine, indolin-2-one, and pseudouridine. KEGG annotation and enrichment analyses of the differential metabolites revealed that flavonoid biosynthesis, amino acids biosynthesis, and arginine and proline metabolism were the main differentially regulated pathways. In conclusion, the petals of C. sativus are valuable for medicine and foods and have potential utility in multiple areas such as the natural spice, cosmetic, health drink, and natural health product industries.

Türkiye'de Safran (Crocus sativus L.) adı ile satılan bazı örneklerin Farmasötik Botanik açıdan değerlendirilmesi

Crocus sativus L. (Saffron, Fam. Iridaceae) is a medicinal, commercial, historical and economically important plant that has been used in the treatment of various diseases since ancient times. Saffron, which is generally used by the public to add flavor and color to dishes, is difficult to cultivate and harvest due to the use of only dried stigmas and is known as the most expensive spice in the world. The macroscopic and microscopic properties of the samples sold under the name "Saffron" from 10 different cities of Turkey, as well as the drying loss, foreign matter amount determination, total ash amount determination, color density determination, phosphomolybdic acid reaction and elongation determination analysis tests were conducted and their conformity with European Pharmacopoeia standards was investigated. In this study, saffron samples purchased from a local producer in Safranbolu (Standard 1) and grown by ourselves (Standard 2) were used. As a result of our study, it was determined by morphological and microscopic studies that samples 2 and 4 completely belonged to the Carthamus tinctorius L. (Safflower) plant. On the other hand, it has been revealed by the analyzes that the sample number six contains very little saffron stigma and mainly contains Safflower bulk flowers.

Effect of safranal on the response of cancer cells to topoisomerase I inhibitors: Does sequence matter?

Lung and colorectal cancers are among the leading causes of death from cancer worldwide. Although topotecan (TPT), a topoisomerase1 inhibitor, is a first- and second-line drug for lung and colon cancers, the development of drug resistance and toxicity still remain as a major obstacle to chemotherapeutic success. Accumulating evidence indicates increased efficacy and reduced toxicity of chemotherapeutic agents upon combining them with natural products. We aimed to investigate the possible interaction of safranal (SAF), a natural compound obtained from Crocus sativus stigma, with TPT when used in different sequences in colon and lung cancer cell lines. The growth inhibitory effect of the proposed combination given in different sequences was assessed using the colony formation assay. The comet assay, cell cycle distribution, Annexin-V staining, and expression of proteins involved in DNA damage/repair were utilized to understand the mechanism underlying the effect of the combination. SAF enhanced the growth inhibitory effects of TPT particularly when it was added to the cells prior to TPT. This combination increased the double-strand break induction and dysregulated the DNA repair machinery, particularly the tyrosyl-DNA phosphodiesterase 1 enzyme. In addition, the SAF + TPT combination increased the fraction of cells arrested at the G2/M checkpoint as well as enhanced the induction of apoptosis. The current study highlights the status of SAF as a natural product sensitizing the lung and colon cancer cells to the cytotoxic effects of the anticancer drug TPT. In addition, it emphasizes the importance of sequence-dependent interaction which can affect the overall outcome.

Crocin ameliorates peritoneal fibrosis in rat induced by peritoneal dialysis via Wnt5a/β-Catenin pathway

Peritoneal dialysis is used in the treatment of patients with kidney diseases. Long-term peritoneal dialysis could result in peritoneal fibrosis and recurrent peritonitis, thus leading to failure of ultrafiltration. Crocin is a bioactive carotenoid and isolated from stigma of Crocus sativus, and ameliorates pulmonary and myocardial fibrosis. The role of crocin in peritoneal fibrosis was assessed. Firstly, rats model with peritoneal dialysis was treated with 4.25% peritoneal dialysate. Results showed that injection with peritoneal dialysate induced obvious hyperplasia and increased thickness in peritoneum structure. Rats with peritoneal dialysis were injected with increasing concentrations of crocin at 10, 20, or 40 mg/kg. Crocin ameliorated the pathological changes in the peritoneum of peritoneal dialysate-induced rats. Secondly, crocin attenuated peritoneal dialysate-induced decrease of E-cadherin, increase of fibronectin, α-smooth muscle actin (α-SMA), and collagen I. Moreover, crocin enhanced ultrafiltration volume and reduced glucose transport in rats model with peritoneal dialysis. Thirdly, crocin also reduced levels of Interleukin (IL)-1β, Tumor Necrosis Factor-α (TNF –α), and IL-6 in peritoneal tissues of rats model with peritoneal dialysis. Lastly, protein expression of Wnt5a and β-Catenin in rats model with peritoneal dialysis were also downregulated by crocin. In conclusion, crocin exerted anti-inflammatory and anti-fibrotic effects on rats model with peritoneal dialysis through inactivation of Wnt5a/β-Catenin pathway.

Novel Insight into the Volatile Profile and Antioxidant Properties of Crocus sativus L. Flowers.

The current production system of saffron spice generates hundreds of tons of waste. Thus, the aim of this study was to value both saffron and its floral by-products as a source of natural bioactive extracts, studying the in vitro antioxidant capacity, the composition of the volatile fraction by GC-MS/MS, and the determination of crocetins esters by HPLC-PDA. Saffron stigmas and floral by-products showed a high content of polyphenols and different antioxidant properties. Floral bio-residues (tepals, stamens, and styles) presented a high concentration of anthocyanins, and stigmas had high levels of flavonoids, β-carotene, and total crocins. In stigmas, 25 different volatile components were found, with safranal the most relevant. Floral by-products volatile composition consisted of 55 compounds with varying amounts depending on the drying treatment; all the samples presented acetic acid, 2(5H)-furanone, and phenylethyl alcohol. Therefore, saffron stigmas and flower by-products represent a sustainable source of bioactive ingredients for innovative healthy food formulations.

Starch-based sustainable hydrogel loaded with Crocus sativus petals extract: A new product for wound care

The aim of the present study was to valorize Crocus sativus petals, the main waste deriving from saffron stigma harvesting, as source of bioactive molecules to be used in health field. Three different dry extracts were prepared by eco-friendly methods (maceration and ultrasound bath assisted maceration) using saffron petals as raw material and ethanol 70 % either ethanol 96 % as extraction solvents. A preliminary evaluation of the antioxidant activity (measured by ABTS*+, DPPH* and FRAP) highlighted that the most suitable extraction solvent is represented by ethanol 70 %. By in vitro studies on keratinocytes emerged that the extract obtained by maceration (rich in gallic and chlorogenic acids) stimulates their growth in a safe concentration range (0.02–0.4 mg/mL) suggesting a potential application in skin diseases such as superficial wounds.Due to the low manageability, the extract was firstly supported on corn starch powder particles and then formulated as starch gel. The obtained formulation showed both suitable rheological properties and spreadability necessary for an easy and pain free application on damaged skin. Moreover, in vitro microbiological studies of starch gel demonstrated antimicrobial activity toward S. epidermidis and self-preserving capacity.

Saffron and its active ingredients against human disorders: A literature review on existing clinical evidence.

Saffron, the stigmas of Crocus sativus L., has been mentioned extensively in the traditional reference texts as a herbal medicine. Many clinical trials have been conducted on this valuable herbal substance and its main constituents following numerous cellular and animal assessments. In the present review, we have collected almost all of these clinical studies to clarify how much knowledge has clinically been achieved in this field so far and which scientific gaps are needed to be filled by more studies. A comprehensive literature review was conducted through a two-round search. First, we performed a general search for identifying the human disorders against which saffron was studied. Then, we searched specifically for the combination of saffron keywords and each disease name. Scientific databases including Scopus, PubMed, and Web of science were used for this search. Studies were collected through electronic databases from their inception up to August 2021. The largest number of these clinical studies represent the investigations into saffron efficacy in different neurological and mental disorders, particularly depression. This substance has clinically revealed significant protective effects against various types of depression, age-related macular degeneration, and allergic asthma. In some cases, such as sexual dysfunction, cognitive and metabolic disorder, the effects of saffron are still clinically open to dispute, or there are limited data on its positive influences. Overall, saffron and its constituents have promising effects on human disorders; however, it needs more clinical evidence or meta-analyses to be confirmed.

Replacing chemical fertilizers with organic and biological ones in transition to organic farming systems in saffron (Crocus sativus) cultivation

To evaluate the response of saffron to animal manure, and biological and chemical fertilizer in an arid climate, an experiment was performed as split plots based on a randomized complete blocks design with three replications during three consecutive crop growth seasons (2015–2018) at the Research Farm of University of Gonabad, Iran. The experimental treatments included application (60 t ha−1) and non-application (control) of manure as the main plot and the use of biosulfur (5 kg ha−1), biophosphate (3 L ha−1), nitroxin (3 L ha−1), chemical fertilizer (150, 100, and 100 kg ha−1 of urea, triple superphosphate, and potassium sulfate, respectively), and no fertilizer application (control) as the sub-plot. The results showed a highly significant response of the quantitative traits of saffron to the application of manure, which increased the leaf, flower, and corm indices of saffron by a mean of 15.1–35.7% than control. The interaction effect of manure with biological and chemical fertilizers for leaf, flower, and weeds indices of saffron was significant. There was no significant difference between the interaction treatments of manure and chemical fertilizer with nitroxin and biophosphorus fertilizers in most of the mentioned traits in the three experiment years. The simultaneous application of these fertilizers increased the average by about 60, 105, 135, 110, 165, and 55% of the leaf dry weight, the number of flowers, fresh flower yield, dry flower yield, dry stigma yield, and weed dry weight of saffron, respectively as compared to control. There was no significant difference between the chemical fertilizer with nitroxin or biophosphate in terms of the effect on the traits related to saffron corm so the use of these fertilizers, as compared to control, increased replacement corm weight, replacement corm size, and bud number per corm by, respectively, about 35, 60, and 40% on average. The chemical and biological fertilizers improved the content of crocin, picrocrocin, and safranal of saffron stigma. The best results were obtained from the use of chemical fertilizers, although no significant difference was observed between this treatment and the nitroxin and biophosphate treatments. Overall, the results of this three-year experiment show a very high response of the saffron plant to the simultaneous use of manure and biological fertilizers and, therefore, it is possible to replace chemical fertilizers with organic and biological fertilizers in saffron cultivation to implement organic agriculture and achieve acceptable quantitative and qualitative yields in areas similar to the experiment location.

Changes of Crocin and Other Crocetin Glycosides in Saffron Through Cooking Models, and Discovery of Rare Crocetin Glycosides in the Yellow Flowers of Freesia Hybrida.

Crocetin glycosides such as crocin are noted as functional food materials since the preventive effects of crocin have been reported against chronic disease and cancer. However, it is unclear how these apocarotenoids are structurally changed through cooking for our intake. We examined such changes in crocetin glycosides (crocin, tricrocin, and crocin-3) contained in saffron (stigmas of Crocus sativus) through cooking models. These glycosides were almost kept stable in boiling for 20 min (a boiled cooking model), while hydrolysis of the ester linkage between glucose and the crocetin aglycone occurred in a grilled cooking model (180°C, 5 min), along with a 13-cis isomerization reaction in a part of crocetin subsequently generated. We further here revealed that the yellow petals of freesia (Freesia x hybrida) with yellow flowers accumulate two unique crocetin glycosides, which were identified to be crocetin (mono)neapolitanosyl ester and crocetin dineapolitanosyl ester. A similar result as above was obtained on their changes through the cooking models. Utility applications of the freesia flowers as edible flowers are also suggested in this study. Additionally, we evaluated singlet oxygen (1O2)-quenching activities of the crocetin glycosides contained in saffron and freesia, and crocetin and 13-cis crocetin contained in the grilled saffron, indicating that they possessed moderate 1O2-quenching activities (IC50 24–64 μM).

Photocatalytic activity of green synthesized cadmium sulfide quantum dots on the removal of RhB dye and its cytotoxicity and antibacterial studies

Presence of inorganic pollutants in water reservoirs is the treating factor for human health and environment. Semiconductor quantum dots (QDs) has been regarded as one of the most efficient nanoparticles for their enhanced photocatalytic activity. Medicinal plants are the safe sources to provide green template for biosynthesis of inorganic nanoparticles such as quantum dots. In order to determine the photocatalytic and biological application of cadmium sulfide quantum dots (CdS QDs), a biosynthesis approach was employed using saffron (Crocus sativus L.) stigma extract as the green reaction substrate. The biosynthesis process was evaluated at different pH condition to obtain the most efficient CdS QDs. Characterization of prepared CdS QDs were determined through UV–vis and fluorescence spectroscopy, FTIR and TEM analysis. The obtained results showed well dsispersed and uniform QDs during green synthesis at the optimum condition. The absorption and electrical properties of green synthesized CdS QDs showed the lowest energy bandgap of 2.4 at pH 11. Photocatalytic activity of CdS QDs on Rhodamine B degradation showed 92% degradation after 80 min under UV light irradiation. The antibacterial and cell cytotoxicity of green synthesized CdS QDs were assayed by disk diffusion and MTT assays respectively. Obtained results showed significant antibacterial effect of CdS QDs against gram-positive and gram-negative bacteria including B. subtilis (90%) and E. coli (96%) respectively. Moreover, cytotoxicity of prepared CdS Qds through MTT assay indicated 79% apoptosis induction on MCF-7 breast cancer cells.

Crocus sativus L. Petal Extract Inhibits Inflammation and Osteoclastogenesis in RAW 264.7 Cell Model.

The dried stigmas of Crocus sativus L. (Iridaceae) are traditionally processed to produce saffron, a spice widely used as a food coloring and flavoring agent, which is important in the pharmaceutical and textile dye-producing industries. The labor-intensive by-hand harvesting and the use of only a small amount of each flower cause saffron to be the most expensive spice in the world. Crocus sp. petals are by-products of saffron production and represent an interesting raw material for the preparation of extracts intended for health protection in the perspective of a circular economy. In the present study, ethanolic extract from Crocus sativus L. petals (Crocus sativus L. petal extract, CsPE) was tested on macrophages by in vitro models of inflammation and osteoclastogenesis. The extract was found to be endowed with anti-inflammatory activity, significantly reducing the nitric oxide production and IL-6 release by RAW 264.7 murine cells. Moreover, CsPE demonstrated an anti-osteoclastogenic effect, as revealed by a complete inhibition of tartrate-resistant acid phosphatase (TRAP)-positive osteoclast formation and a decreased expression of key osteoclast-related genes. This study, which focuses on the macrophage as the target cell of the bioactive extract from Crocus sativus L. petals, suggests that the petal by-product of saffron processing can usefully be part of a circular economy network aimed at producing an extract that potentially prevents bone disruption.

An Overview of Molecularly Imprinted Polymers Embedded with Quantum Dots and Their Implementation as an Alternative Approach for Extraction and Detection of Crocin

AbstractMolecularly imprinting polymer (MIPs) is an emerging technology that could provide an alternative path to produce receptor‐like binding sites. The technology involves complex bond formation between target molecules (templates), and functional monomers can occur insolvent. MIPS embedded with Quantum Dots (QDs) as a sensor is used for extraction and detection of various compounds. QDs have fluorescence properties and electronic characteristics such as comprehensive and continuous absorption spectra, narrow emission spectra, and high light resistance. Crocin is the major metabolite obtained from saffron (Crocus sativus). It is widely used as an antioxidant, antidiabetic, antitumor, and anticancer agent. According to recent studies, crocin has been used for drug delivery to treat various diseases. Therefore, the isolation, extraction and detection of crocin from saffron are of prime importance. Developing a selective extraction and detection technique for crocin would assist in its separation and purification easily from saffron stigmas. MIPs encapsulated with QDs is a promising tool for the extraction and detection of various compound, and this technique could also be employed for crocin. In this review, a brief summary of MIPs, QDs and MIP‐QDs sensors has been summarised. Moreover, an effort has been made to review how MIPs encapsulated with QDs can be used as the best alternative for efficiently extracting and detecting crocin.

Phytochemical profiling, antibacterial and antioxidant properties of Crocus sativus flower: A comparison between tepals and stigmas

Abstract Several studies have demonstrated that stigmas of Crocus sativus contain several bioactive compounds with potential health-promoting properties. However, during the processing of stigmas, large amounts of floral bio-residues are normally discarded as by-products. In this study, using untargeted metabolomics, the comprehensive phytochemical composition of C. sativus stigma and tepals was investigated. Moreover, the antibacterial and anti-biofilm properties of the extracts of C. sativus stigmas and tepals were compared. The study was carried out using two methicillin-resistant staphylococcal reference strains (i.e., Staphylococcus aureus ATCC 43300 and Staphylococcus epidermidis ATCC 35984), representing important Gram-positive biofilm-forming human pathogens. The antibacterial properties were correlated with total polyphenol content, total terpenoid content, and in vitro antioxidant properties of tepals and stigmas. The results demonstrated that stigma and tepal extracts, at the sub-toxic concentrations, were able to interfere with biofilm formation by ATCC 43300 and ATCC 35984. Besides, the higher antibacterial activity of tepals than stigmas was associated with higher levels of phycompounds. Therefore, our results demonstrated that C. sativus stigmas and bio-residues, such as tepals, are potential antioxidant sources and good candidates as antibacterial agents to prevent biofilm formation. Taken together, these findings showed that C. sativus could be used as functional ingredient by the food and pharmaceutical industries.

Safety Assessment and Pain Relief Properties of Saffron from Taliouine Region (Morocco).

Saffron is the most expensive spice in the world. In addition to its culinary utilization, this spice is used for medicinal purposes such as in pain management. In this study, the analgesic activity of Crocus sativus stigma extract (CSSE) was evaluated in rodents and its possible physiological mechanism was elucidated. The anti-nociceptive effect of CSSE was evaluated using three animal models (hot plate, writhing, and formalin tests). The analgesic pathways involved were assessed using various analgesia-mediating receptors antagonists. The oral administration of CSSE, up to 2000 mg/kg, caused no death or changes in the behavior or in the hematological and biochemical blood parameters of treated animals nor in the histological architecture of the animals’ livers and kidneys. CSSE showed a central, dose-dependent, anti-nociceptive effect in response to thermal stimuli; and a peripheral analgesic effect in the test of contortions induced by acetic acid. The dual (central and peripheral) analgesic effect was confirmed by the formalin test. The anti-nociceptive activity of CSSE was totally or partially reversed by the co-administration of receptor antagonists, naloxone, atropine, haloperidol, yohimbine, and glibenclamide. CSSE influenced signal processing, by the modulation of the opioidergic, adrenergic, and muscarinic systems at the peripheral and central levels; and by regulation of the dopaminergic system and control of the opening of the ATP-sensitive K+ channels at the spinal level. The obtained data point to a multimodal mechanism of action for CSSE: An anti-inflammatory effect and a modulation, through different physiological pathways, of the electrical signal generated by the nociceptors. Further clinical trials are required to endorse the potential utilization of Moroccan saffron as a natural painkiller.

Antioxidant Activity, Metal Chelating Ability and DNA Protective Effect of the Hydroethanolic Extracts of Crocus sativus Stigmas, Tepals and Leaves.

The present study investigated the antioxidant activity, metal chelating ability and genoprotective effect of the hydroethanolic extracts of Crocus sativus stigmas (STG), tepals (TPL) and leaves (LV). We evaluated the antioxidant and metal (Fe2+ and Cu2+) chelating activities of the stigmas, tepals and leaves of C. sativus. Similarly, we examined the genotoxic and DNA protective effect of these parts on rat leukocytes by comet assay. The results showed that TPL contains the best polyphenol content (64.66 µg GA eq/mg extract). The highest radical scavenging activity is shown by the TPL (DPPH radical scavenging activity: IC50 = 80.73 µg/mL). The same extracts gave a better ferric reducing power at a dose of 50 µg/mL, and better protective activity against β-carotene degradation (39.31% of oxidized β-carotene at a 100 µg/mL dose). In addition, they showed a good chelating ability of Fe2+ (48.7% at a 500 µg/mL dose) and Cu2+ (85.02% at a dose of 500 µg/mL). Thus, the antioxidant activity and metal chelating ability in the C. sativus plant is important, and it varies according to the part and dose used. In addition, pretreatment with STG, TPL and LV significantly (p < 0.001) protected rat leukocytes against the elevation of percent DNA in the tail, tail length and tail moment in streptozotocin- and alloxan-induced DNA damage. These results suggest that C. sativus by-products contain natural antioxidant, metal chelating and DNA protective compounds, which are capable of reducing the risk of cancer and other diseases associated with daily exposure to genotoxic xenobiotics.

FC021: Saffron-Derived Bioactive Molecules and Their in-vitro Activity on Kidney and Bladder Tumoral Cells

Abstract BACKGROUND AND AIMS Saffron is the dried stigmas of Crocus sativus L. Crocus sativus L belongs to the family of Iridaceae and is mainly cultivated in several countries with mild and dry climates. It is one of the most expensive spices in the world and the main reason for its great cost is that saffron is still cultivated and harvested as it has been for millennia by hand. Saffron's name is derived from the Arab word for yellow, a name reflecting the high concentration of carotenoid pigments present in the saffron flowers’ stigmas which contribute most to the colour profile of this spice. Saffron contains numerous bioactive molecules known for their antioxidant activities, but also some such as picrocrocin, crocetin and Safranal (Sfr), Crocin (Crc) which are known to possess the ability to inhibit tumoral growth by inducing apoptosis. The aim of this study is to understand whether these four saffron-derived bioactive molecules display tumoral growth inhibition activity on kidney and bladder cancer cell lines. METHOD Saffron samples used for this purpose possess both different origins (Tuscany and Lombardy) and harvesting years (T_2017, T_2018, L_2017, L_2018) and, in addition, Sfr and Crc standard samples were also tested. Identification of Crocins, Safranal, Picrocrocin and Flavonoids was carried out by HPLC/DAD and HPLC/MS analysis. The identity of polyphenols was ascertained using data from HPLC/DAD analyses by comparison and combination of their retention times and UV/Vis spectra with those of authentic standards and previously reported data (Vignolini et al., 2008). The extract characterization is summarized in the table below. The extracts’ effects on cancer cell viability were evaluated through MTT assays and the tumoral cell lines chosen were Caki-1 and 786-O for the kidney and RT112 and RT4 for the bladder. MDCK cells were used as a healthy kidney model. RESULTS In the presence of both pure Sfr and Crc, post-hoc tests have shown a time and concentration-dependent decrease in viability for all the cancerous models in analysis, with more visible results observable for Crc, while having no effect on MDCK viability. Being Crc is the most represented molecule in all the extracts, before performing MTT assays, a normalization of this compound's concentration was carried out. L_2017, which contained the highest Sfr concentration among all the extracts tested, showed the highest inhibiting activity. Hypothesizing that Sfr was the molecule responsible for the decrease in cancer cell viability, all extracts and Sfr and Crc standards were tested on RT112 cells after normalizing the contents of these two molecules for each condition. We observed that Crc and Sfr alone and the Sfr-Crc mix had a small impact on cancer cell viability when compared with the effects of whole extracts. CONCLUSION Whole extracts demonstrated a higher impact on cancer cell viability compared with the standards tested both singularly and combined. This study, therefore, recognizes that numerous molecules in combination with Sfr and Crc have a role in reducing tumoral cell viability, demonstrating that the phyto-complex may possess higher therapeutic effects.

Analysis of Crocetins and Safranal Variations in Saffron (Crocus sativus) Stigma Samples and Dietary Supplements Using HPLC/UHPLC-PDA-MS: Chemical Profiling and Chemometric Analysis Using LC-QToF

Analysis of Crocetins and Safranal Variations in Saffron (Crocus sativus) Stigma Samples and Dietary Supplements Using HPLC/UHPLC-PDA-MS: Chemical Profiling and Chemometric Analysis Using LC-QToF

Does nutrient and irrigation managements alter the quality and yield of saffron (Crocus sativus L.)?

Increasing the saffron quantity and quality is essential and should be considered to achieve the desired performance. In this regard, to study the effect of nutrient and irrigation management on the quality and yield of saffron, an experiment was carried out at the research station, the Ferdowsi University of Mashhad, Iran, in 2013–2015. The study was performed as a split-split plot based on a Randomized Complete Block Design with 18 treatments. Experimental factors were: 1- superabsorbent (S.A.) [application and non-application (non-S.A.)], 2- irrigation intervals [every two, three, and four weeks], and 3- nutrient management [humic acid, mycorrhiza (Glomus intraradices) and control]. Among all treatments, the application of S.A. increased on average 61% fresh saffron flower and dry stigma yields in both growing seasons. S.A. + four weeks irrigation intervals + humic acid were increased number of flower, dry stigma yields (175.33 per m2 and 7.472 kg.ha-1, respectively), and water productivity of irrigation of saffron stigma (2.138 g.ha-1) in 2015. The use of S.A. and humic acid compensated for water shortage and increased stigmas' yield. Furthermore, the maximum L observed in S.A. + three weeks irrigation intervals + humic acid (32.44). Among all treatments, the lowest amount of b value and Hue was observed in shorter irrigation intervals; so, application of S.A. + two weeks irrigation intervals + humic acid resulted in the minimum amount of Hue (−0.1783) that was a desirable trait. The maximum crocin and picrocrocin obtained in S.A. + three weeks irrigation intervals + mycorrhiza (225.9 A4401% and 86.58 A2541%, respectively) and the highest safranal content observed in S.A. + four weeks irrigation intervals + humic acid (47.03 A3301%). In general, it seems that the application of humic acid, mycorrhiza, S.A., and longer irrigation intervals can significantly increase the quality and yield of saffron.