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Objectives To prepare a lyophilized saffron aqueous extract (SFE) and determine its chemical profile and serum and tissue pharmacokinetics after intravenous and oral administration to C57/Bl6J mice. Methods Lyophilized SFE was prepared, characterized using semi-preparative HPLC and NMR analysis, and stability studies at room temperature, and was quantified for crocin content with an HPLC-PDA method. After intravenous and oral administration of SFE (60 mg/kg, reconstituted with water for injection) to C57/Bl6J mice, crocetin (derived from in vivo crocin hydrolysis) serum and tissue levels (unconjugated and total) were measured with an HPLC-PDA method and subjected to compartmental and non-compartmental PK analysis. Key findings Saffron aqueous extract was rich in all-trans-crocin (27.8 ± 0.1% w/w) and stable for more than 15 months. One-compartment PK model described crocetin's (unconjugated) kinetics after intravenous administration of SFE, while a first-order kinetic parameter described the rate of crocetin biotransformation to crocetin metabolite (conjugated). Α οne-compartment PK model with first-order absorption described crocetin and crocetin's metabolite kinetics after SFE oral administration. Relative oral bioavailability was calculated at 1.17 for total crocetin. Tissue NCA PK analysis revealed extensive crocetin distribution to liver and kidneys. Conclusions SFE is a stable lyophilized extract rich in all-trans-crocin. The PK study allowed the estimation of basic PK parameters and the bioavailability of SFE’s main bioactive component, crocetin, after peros administration.

Ultraviolet B (UVB) radiation induces the production of reactive oxygen species (ROS), resulting in the aging of dermal fibroblasts. Crocin, a bioactive constituent of Crocus sativus, possesses anti‑oxidation effects. The purpose of the present study was to evaluate the protective effect of crocin on UVB‑induced dermal fibroblast photoaging. Human dermal fibroblasts were isolated and cultured with different concentrations of crocin prior to and following exposure to UVB irradiation. The senescent phenotypes of cells were evaluated, including cell proliferation, cell cycle, senescence‑associated β‑galactosidase (SA‑β‑gal) expression, intracellular ROS, expression of antioxidant protein glutathione peroxidase 1 (GPX‑1) and extracellular matrix protein collagen type 1 (Col‑1). Crocin rescued the cell proliferation inhibited by UVB irradiation, prevented cell cycle arrest and markedly decreased the number of SA‑β‑gal‑positive cells. In addition, crocin reduced UVB‑induced ROS by increasing GPX‑1 expression and other direct neutralization effects. Furthermore, crocin promoted the expression of the extracellular matrix protein Col‑1. Crocin could effectively prevent UVB‑induced cell damage via the reduction of intracellular ROS; thus, it could potentially be used in the prevention of skin photoaging.

With nearly 140 α-glycosidases in 14 different families, plants are well equipped with enzymes that can break the α-glucosidic bonds in a large diversity of molecules. Here, we introduce activity-based protein profiling (ABPP) of α-glycosidases in plants using α-configured cyclophellitol aziridine probes carrying various fluorophores or biotin. In Arabidopsis (Arabidopsis thaliana), these probes label members of the GH31 family of glycosyl hydrolases, including endoplasmic reticulum-resident α-glucosidase-II Radial Swelling3/Priority for Sweet Life5 (RSW3/PSL5) and Golgi-resident α-mannosidase-II Hybrid Glycosylation1 (HGL1), both of which trim N-glycans on glycoproteins. We detected the active state of extracellular α-glycosidases such as α-xylosidase XYL1, which acts on xyloglucans in the cell wall to promote cell expansion, and α-glucosidase AGLU1, which acts in starch hydrolysis and can suppress fungal invasion. Labeling of α-glycosidases generates pH-dependent signals that can be suppressed by α-glycosidase inhibitors in a broad range of plant species. To demonstrate its use on a nonmodel plant species, we applied ABPP on saffron crocus (Crocus sativus), a cash crop for the production of saffron spice. Using a combination of biotinylated glycosidase probes, we identified and quantified 67 active glycosidases in saffron crocus stigma, of which 10 are differentially active. We also uncovered massive changes in hydrolase activities in the corms upon infection with Fusarium oxysporum using multiplex fluorescence labeling in combination with probes for serine hydrolases and cysteine proteases. These experiments demonstrate the ease with which active α-glycosidases and other hydrolases can be analyzed through ABPP in model and nonmodel plants.

Rheumatoid arthritis (RA) is a systemic autoimmune disease, characterized by the irreversible joint destruction resulted from the attack of inflammatory cells to the joints. Recent studies demonstrated that crocin is able to alleviate arthritis and suppress inflammatory responses, implying crocin as a potential promising antiarthritic agent. In this study, we confirmed the effect of crocin on RA and revealed its underlying mechanism by measuring lipopolysaccharides (LPS)-stimulated cytokine production in presence or absence of crocin. The effect of crocin was also tested in vivo using a mouse model of collagen-induced arthritis (CIA). It was found that crocin significantly repressed the LPS-induced expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 in human fibroblast-like synoviocytes (FLS). We tested the effect of crocin on nuclear factor-kappa B (NF-κB) signaling and observed that cells pre-treated with 500 μM of crocin exhibited lower levels of LPS-induced p-IκBα, p-IκB kinase (IKK) α/β, and p65 expression than those of untreated cells. In addition, we found when cells were stimulated with IKKβ, crocin pre-treatment showed significantly inhibitory effect on the luciferase activity of IL-1β. In vivo results also showed that crocin treatment dramatically reduced plasma levels of TNF-α, IL-1β, and IL-6 in CIA mice. Crocin is efficient to suppress the productions of TNF-α, IL-6, and IL-1β by blocking NF-κB signal activation through its interaction with IKK, suggesting that crocin could be an efficient treatment for RA.

Diabetes mellitus is a metabolic disease that manifested as hyperglycemia due to the defect in secretion or function of insulin. Studies have shown that saffron and its derivatives cause a significant reduction in plasma glucose levels in experimental models. The purpose of this study was to investigate the effect of the saffron extract on fasting plasma glucose (FPG), glycated hemoglobin level (HbA1c), lipid profile, liver enzymes, and renal function tests in type 2 diabetic patients. In this double-blind randomized clinical trial, 64 type 2 diabetic patients who were on oral anti-diabetic drugs were examined. Participants received either 15 mg of saffron or placebo capsules (two pills per day) for 3 months. Anthropometric indices, dietary intake, FPG, HbA1c, lipid profiles, liver enzymes (ALT, AST, ALP), and renal function (BUN, Cr.) tests were measured pre and post intervention after 3 months. Independent t test and paired t test were used for data analysis. After 3-months intervention, mean difference of FPG, Cholesterol, LDL-c, and LDL/HDL ratio between two groups showed significant reduction(p 0.0001), but HbA1c, HDL-C, API, TG showed no significant differences (p 0.05). In saffron group, FPG, HbA1c, cholesterol, LDL-c, and LDL/HDL ratio decreased significantly after 3-months intervention compare with baseline (p 0.0001).

In saffron, the cleavage of zeaxanthin by means of CCD2 generates crocetin dialdehyde, which is then converted by an unknown aldehyde dehydrogenase to crocetin. A proteome from saffron stigma was released recently and, based on the expression pattern and correlation analyses, five aldehyde dehydrogenases (ALDHs) were suggested as possible candidates to generate crocetin from crocetin dialdehydes. We selected four of the suggested ALDHs and analyzed their expression in different tissues, determined their activity over crocetin dialdehyde, and performed structure modeling and docking calculation to find their specificity. All the ALDHs were able to convert crocetin dialdehyde to crocetin, but two of them were stigma tissue-specific. Structure modeling and docking analyses revealed that, in all cases, there was a high coverage of residues in the models. All of them showed a very close conformation, indicated by the low root-mean-square deviation (RMSD) values of backbone atoms, which indicate a high similarity among them. However, low affinity between the enzymes and the crocetin dialdehyde were observed. Phylogenetic analysis and binding affinities calculations, including some ALDHs from Gardenia jasmonoides, Crocus sieberi, and Buddleja species that accumulate crocetin and Bixa orellana synthetizing the apocarotenoid bixin selected on their expression pattern matching with the accumulation of either crocins or bixin, pointed out that family 2 C4 members might be involved in the conversion of crocetin dialdehyde to crocetin with high specificity.

The specific PCR primer was designed base on ITS2 sequence in GenBank, and we developed a SYBRGreen real-time fluorescence quantitative PCR system for identification of Crocus sativus and Carthamus tinctorius source. Compared with Chinese herbal medicine DNA barcode technique, this method showed characteristics of shorter time, higher specificity and sensitivity. Using this method to detect 15 samples, 4 were C. sativus, 8 were C. tinctorius, and the other 3 samples were none of them. The result was in accordance with Chinese herbal medicine DNA barcode. This study lays the foundation for identification of related Chinese medical materials.

Hypoxia as one of the most common clinical disturbances in pregnancy period can cause destructive changes in motor sensory cortex and can lead to imperfect organization in motor reactions. Crocin, a water-soluble carotenoid, is the most active ingredients of saffron and a lot of studies declare its positive effectiveness on improving motor activity. Since the hypoxia intensity affects its malicious amount on movement, in this paper, we have studied the effect of crocin in three maternal hypoxia protocols with different oxygen intensities on motor activity and balance in rat offspring. In this experiment, female rats (Wistar) were used on the 20th day of pregnancy. The rats were randomly divided into eight experimental groups: sham, crocin, hypoxia with three different intensities: 10% oxygen and 90% nitrogen for 1 h (hypoxia-ɪ), 7% oxygen and 93% nitrogen for 1 h (hypoxia-ɪɪ), 7% oxygen and 93% nitrogen for 3 h (hypoxia-ɪɪɪ) and treated-crocin hypoxia groups. To produce hypoxia, pregnant rats were placed in a hypoxia box. In crocin group, rat offspring received 30 mg/kg crocin via IP injection at P14–28. Control group also received saline injection at the same time. Finally, balance and motor activity in offspring were measured respectively by rotarod and open-field devices. Results showed that motor activity significantly decreased in hypoxia-ɪɪɪ group as compared with sham group (p

Introduction: Resin composite restorative materials can mimic the natural color and shade of the tooth. However, exogenous colorants from food and drinks can stain them due to adsorption. The influence of Indian food colorants and spices on resin composite restorations has not been evaluated extensively. Aim: This study aims to evaluate the staining capacity of microhybrid and nanohybrid resin-based composites, to saffron extract, tandoori powder, and turmeric powder. Subjects and Methods: Forty samples of microhybrid (Kulzer Charisma) and nanohybrid (3M Filtek Z350) resin composites were prepared using an acrylic template of dimension 5 mm × 3 mm. They were randomly divided into four groups and immersed into solutions of saffron extract, tandoori powder, and turmeric powder. Distilled water was used as the control group. Color values (LFNx01, aFNx01, bFNx01) were measured by colorimeter using the CIE LFNx01aFNx01bFNx01 system before and after 72 h of immersion. Color differences ΔEFNx01ab were statistically analyzed. Statistical Analysis Used: Two-way ANOVA and post-hoc Tukey (honest significant difference) test were done using IBM SPSS Statistics for Windows, Version 19.0. Armonk, NY: IBM Corp. Results and Discussion: All the immersion media changed the color of the resin composites to varying degrees. However, turmeric solution showed the maximum mean color variation ΔEFNx01ab of 14.8 ± 2.57 in microhybrid resin composites and 16.8 ± 3.50 in nanohybrid resin composites. Conclusion: Microhybrid and nanohybrid resin composites tend to stain to Indian food colorants, especially to turmeric powder.

Background Chronic obstructive pulmonary disease (COPD) has been emerging as a great health problem in world. Cigarette smoke is known to cause oxidative stress and deplete glutathione (GSH) levels. Nuclear erythroid-related factor 2 (Nrf2) is involved in transcriptional regulation of glutamate-cysteine ligase catalytic subunit (GCLc). Antioxidant compounds may be of therapeutic value in monitoring disease progression. Crocin demonstrates antioxidant and anti-inflammatory functions. The aim of this study was to investigate the protective role of crocin against CSE-mediated oxidative stress, inflammatory process, Nrf2 modifications and impairment of cardiac function in rats with COPD. Methods Eighty rats were divided into four groups: Control, Cigarette smoke exposure (CSE), Crocin, Crocin+CS. Each group was divided into the two parts: 1) to evaluate lung inflammatory and oxidative process, 2) to evaluate the effect of Cigarette smoke induced-lung injuries on cardiac electrocardiogram (such as heart rate and QRS complex) and hemodynamic parameters (such as perfusion pressure and left ventricular developed pressure). Results CSE rats showed a significant increase in cotinine concentration (17.24 ng/ml), and inflammatory parameters and a decrease in PO2 (75.87 mmHg) and expression of PKC (0.86 fold), PI3K (0.79 fold), MAPK (0.87 fold), Nrf2 (0.8 fold) and GCLc (0.75 fold) genes, antioxidant activity, and finally cardiac abnormalities in electrocardiogram and hemodynamic parameters. Co-treatment whit crocin could restore all these values to normal levels. Conclusions CS induced-COPD in rat model provides evidence that chronic CS exposure leads to lung injury and mediated cardiac dysfunction. Crocin co-treatment by modulating of Nrf2 pathway protected lung injury caused by COPD and its related cardiac dysfunction. In this study, we showed the importance of Nrf2 activators as a therapeutic target for the development of novel therapy for lung oxidative injuries.

Permafrost is a major component of the Earth climatic system. Global warming provokes the degradation of permafrost which favors biogeochemical activity in Arctic soils. The decomposition of organic matter increases and results in the release of high amounts of greenhouse gases (CO2 and CH4) to the atmosphere. By amplifying the greenhouse effect induced by human activities, this phenomenon may constitute one of the strongest positive feedbacks on global warming. Predicting these effects requires to study the evolution of the permafrost thermal regime and the factors governing it. The snowpack, because of its insulating effect, modulates the heat fluxes between permafrost and atmosphere most of the year. The snow insulating capacity depends on snow height and thermal conductivity. These two variables are highly dependent on climatic conditions and on the presence of vegetation. Here we monitor the snow and soil physical properties at a high Arctic site typical of herbaceous tundra (Bylot Island, 73°N), and at a low Arctic site situated at the limit between shrub and forest tundra (Umiujaq, 56°N). We use data from automatic measurement stations and manual measurements. A special attention is given to the snow thermal conductivity because very few data are available for Arctic regions. Results are interpreted in relation to vegetation type and atmospheric conditions. The numerical coupled model ISBA-Crocus is then used to simulate snow and soil properties at our sites. Results are compared to field data in order to evaluate the model capacity to accurately simulate the permafrost thermal regime.We managed to describe atmosphere-snow-vegetation interactions that shape the structure of Arctic snowpacks. Wind and the snow redistribution it induces are fundamental parameters governing snow height and thermal conductivity. A high vegetation cover (i.e. shrubs and forest) traps blowing snow and shields it from wind compaction. Vegetation growth thus favors the formation of an insulating snowpack which slows down or even prevents soil freezing. Furthermore, the shrubs woody structure supports the snow mass and prevents the resulting compaction of bottom snow layers. Thus sheltered, snow in shrubs develops a high insulating capacity which delays soil freezing. Continued atmospheric cooling increases the thermal gradient in the snow, maintaining large water vapor transfers from the soil and the snow basal layers to upper layers and atmosphere. The growth of depth hoar, enhanced by the large thermal gradient and the low snow density, results in the formation of highly insulating snow layers thus constituting a positive feedback loop between soil temperature and snow insulation. As long as the soil stays relatively warm, depth hoar growth persists. Finally, if warm spells occur in autumn, they can trigger the partial melting of the early snowpack which can cancel or temporarily reverse the insulating effect of snow-vegetation interactions. A frozen snow surface prevents snow drifting and its redistribution. The presence of highly conductive refrozen layers facilitates soil cooling and reduces the thermal gradient. An early snowpack affected by melting is thus less insulative which could hamper Arctic soil warming. Simulation results show that these different effects are not correctly represented in snow models. Errors in the estimated snow thermal conductivities are particularly problematic as they highly affect the simulation of soil freezing. Given the area of permafrost-affected regions, these errors on Arctic snow modelling could significantly impact climate simulations and the global warming projections., Le pergélisol est une composante majeure du système climatique terrestre. Avec le réchauffement du climat, la dégel du pergélisol profite à l'activité biochimique qui décompose davantage de matière organique dans les sols arctiques et la rejette dans l'atmosphère sous forme de gaz à effet de serre (CO2, CH4). Ce phénomène pourrait constituer une rétroaction climatique positive majeure. Prédire ces effets nécessite d'étudier l'évolution du régime thermique du pergélisol ainsi que des facteurs qui l'influencent. Le manteau neigeux, de par son pouvoir isolant, contrôle les échanges de chaleur entre le sol et l'atmosphère une grande partie de l'année. Le flux de chaleur à travers la neige dépend de la hauteur du manteau neigeux et de la conductivité thermique des couches de neige qui le constituent. Ces deux variables sont elles-même très dépendantes des conditions climatiques et de la présence de végétation. Nous réalisons ici le suivi des propriétés de la neige et du sol d'un site haut arctique de toundra herbacée (Île Bylot, 73N), et d'un site bas arctique à la frontière de la toundra arbustive et forestière (Umiujaq, 56N). Nous utilisons les données issues de stations de mesure automatiques complétées par des mesures manuelles. Une attention particulière est portée sur la conductivité thermique de la neige, car peu de données sont disponibles pour les régions arctiques. Le modèle numérique couplé ISBA-Crocus est ensuite utilisé pour simuler les propriétés de la neige et du sol des deux sites étudiés. Les résultats sont comparés aux mesures de terrain afin d'évaluer la capacité du modèle à simuler le régime thermique des sols arctiques.Nous avons pu caractériser les interactions atmosphère-neige-végétation qui façonnent la structure des manteaux neigeux arctiques. Le vent et la redistribution de neige qu'il induit sont des paramètres fondamentaux qui déterminent la hauteur et la conductivité thermique de la neige. Un couvert végétal haut et dense (arbustes, arbres) piège la neige soufflée et l'abrite du tassement éolien. De plus, la structure ligneuse des massifs arbustifs soutient la masse de neige et empêche son tassement. Cet abri procure à la neige une capacité d'isolation élevée qui retarde le gel du sol dès les premières accumulations. Le refroidissement atmosphérique se poursuivant, le manteau neigeux peu épais est soumis à un gradient thermique élevé qui provoque d'importants transferts de vapeur d'eau depuis le sol et les couches de neige basales, vers les couches supérieures et l'atmosphère. La croissance de givre de profondeur qui s'opère, favorisée à la fois par le gradient thermique élevé et la faible densité de la neige, aboutit à la formation de couches très isolantes en contact avec la surface du sol. Tant que le sol demeure relativement chaud, la croissance de givre de profondeur perdure. Finalement, des épisodes de fonte peuvent avoir lieu en automne durant la mise en place du manteau neigeux dans les régions arctiques. Le regel de la neige peut rapidement annuler ou même temporairement inverser l'effet isolant des interactions neige-végétation. Une surface de neige gelée ne subit pas l'effet du vent et empêche sa redistribution. La formation de croûtes de regel à forte conductivité thermique accélère le refroidissement du sol. Le manteau neigeux affecté par la fonte au début de l'hiver a donc une capacité d'isolation diminuée qui pourrait entraver le réchauffement des sols arctiques. Nos résultats de simulation montrent que ces différents effets ne sont pas correctement représentés dans les modèles de neige. Les erreurs dans les conductivités thermiques de la neige simulées sont particulièrement problématiques puisqu'elles interviennent lors de la période de gel du sol. Étant donné l'étendue des régions affectées par le pergélisol, ces erreurs sur la modélisation de la neige arctique pourraient significativement affecter les simulations climatiques et les projections de la hausse des températures globales.

The dried stigma of Crocus sativus (saffron) is a worlds precious spice and traditional medicine possessing plenty of pharmacological effects. Because the stigma represents only 7.4% of total weight of the fresh flower, when the stigma is separated from the flowers, large quantities of floral bio-residues are discarded. Accordingly, the utilization of the non-medicinal parts of saffron has drawn peoples attention. Studies showed that the non-medicinal parts of saffron were rich in flavonoids, phenolic acids, polysaccharides, et al. They were proved to have some pharmacological activities, such as antioxidant activity, antifungal activity, cytotoxicity, anti-inflammation and liver protection. The paper is to review the current research progress on the chemical components and pharmacological effects of the non-medicinal parts of saffron, which can serve as a reference for the further research and development of non-medicinal parts of saffron.

Crocus sativus L., is a plant cultivated in many countries of the world. Crocins are among the active constituents of C. sativus and their implication in cognition has been proposed. The present study was designed to investigate in the rat the effects of crocins on distinct recognition memory components (encoding, storage and retrieval). Subsequently, the potential use of crocins as adjunctive agents for the treatment of memory disorders was examined. Thus, the effects exerted by a combination of subthreshold doses of crocins and memantine on recognition memory were evaluated. To assess the effects of compounds on memory, the novel object-recognition task (NORT) was used. In a preliminary study, the influence of the retention time (the delay between the two trials) on the performance of rats was assessed. Rats' recognition memory abilities remained intact up to 6 h, but were extinguished when a delay of 24 h was utilized. Crocins, at any dose tested (5, 15, and 30 mg/kg), did not affect rats' performance, whereas administration of higher doses (15 and 30 mg/kg) reversed delay-dependent deficits in the NORT. The combination of subthreshold doses of crocins (5 mg/kg) and memantine (3 mg/kg) did not influence the performance, but counteracted delay-dependent deficits in the NORT. These findings suggest that crocins counteract natural forgetting and may modulate different aspects of recognition memory, and that the combined use of crocins and memantine might represent a novel strategy to treat memory disorders.

Crocus sativus L. (Saffron) has long been known for multiple target therapeutic uses. The plant metabolism is well investigated and the main metabolites related to saffron organoleptic qualities are crocin, crocetin, picrocrocin, and safranal. Particularly, the most abundant of them, such as crocin and safranal, are investigated for their multiple biological activities and known as potential drugs. We aimed to review the constituent features of the plant, along with its potential therapeutic effects in depression, neurodegenerative diseases, diabetes mellitus, atherosclerosis, cancer, and sexual dysfunction. A systematic literature search was conducted in PubMed, Medline, Scopus, and EMBASE, with particular attention to preclinical and clinical studies. Although saffron and its components showed potential clinical applications, further investigations are necessary to confirm the effective use of "Red Gold" and its real applications in clinical practice.

A change in homeostasis between food intake and energy expenditure is the hallmark of obesity. Many plant-based weight-management products are available in dietary supplement markets with no direct efficacy comparison. In this in vivo acute feed intake study in rats, the appetite suppression efficacy of well-known natural ingredients in the weight-loss market were evaluated. We tested pure caffeine, potato skin extract, Cissus quadrangularis extract, Garcinia cambogia extract, Crocus sativus extract, raspberry ketone isolated from Rubus idaeus, one commercial product (Appetrex), and one novel composition (UP601). Rats treated with potato skin extract, Crocus sativus bulb extract, and Cissus quadrangularis extracts showed statistically significant reduction in food consumption only at the 2-hour timepoint with 44.9%, 34.1%, and 44.3% reductions, respectively, after food provision at an equivalent human dosage of 2 g, 10 g, and 10 g, respectively. Garcinia cambogia fruit extract and raspberry ketone from Rubus idaeus showed statistically significant reduction in food consumption only at the 1-hour timepoint with 33.7% and 79.4% reductions, respectively, after food provision at an equivalent human dosage of 8 g and 5 g, respectively. UP601 and Appetrex were compared at 230 mg/kg. While 88.5%, 73.8%, and 63.1% reductions in food intake were observed for the UP601 treatment group, 64.2%, 27.5%, and 34.7% reductions in food intake were observed for rats treated with Appetrex at 1 h, 2 h, and 4 h after food provision. The composition UP601 demonstrated superior activity in food intake compared to any of the dietary supplements marketed for appetite suppression tested in this study.

Crocins, the red soluble apocarotenoids of saffron, accumulate in the flowers of Crocus species in a developmental and tissue-specific manner. In Crocus sieberi, crocins accumulate in stigmas but also in a distinct yellow tepal sector, which we demonstrate contains chromoplast converted from amyloplasts. Secondary metabolites were analysed by LC-DAD-HRMS, revealing the progressive accumulation of crocetin and crocins in the yellow sector, which were also localized in situ by Raman microspectroscopy. To understand the underlying mechanisms of crocin biosynthesis, we sequenced the C. sieberi tepal transcriptome of two differentially pigmented sectors (yellow and white) at two developmental stages (6 and 8) by Illumina sequencing. A total of 154 million high-quality reads were generated and assembled into 248,099 transcripts. Differentially expressed gene analysis resulted in the identification of several potential candidate genes involved in crocin metabolism and regulation. The results provide a first profile of the molecular events related to the dynamics of crocetin and crocin accumulation during tepal development, and present new information concerning apocarotenoid biosynthesis regulators and their accumulation in Crocus. Further, reveals genes that were previously unknown to affect crocin formation, which could be used to improve crocin accumulation in Crocus plants and the commercial quality of saffron spice.

The dried stigmas of Crocus sativus constitute the saffron, which is considered to be the costliest spice of the world. Saffron is valuable for its constituents, which are mainly apocarotenoids. In order to enhance the production of apocarotenoids, it is imperative to understand the regulation of apocarotenoid biosynthetic pathway. In C. sativus, although the pathway has been elucidated, the information regarding the regulation of the pathwaygenes is scanty. During the present investigation, the characterization of promoters regulating the expression of two important genes i.e. CsPSY and CsUGT was performed. We successfully cloned the promoters of both the genes, which were functionally characterized in Crocus sativus and Nicotiana tabaccum. In silico analysis of the promoters demonstrated the presence of several important cis regulatory elements responding tolight, hormonesand interaction with transcription factors (TFs). Further analysis suggested the regulation of CsPSY promoter by Abscisic acid (ABA) and that of CsUGT by Gibberellic acid (GA). In addition, we also observed ABA and GA mediated modulation in the expression of significant TFs and CsPSY and CsUGT transcripts. Overall, the study addresses issues related to regulation of key genes of apocarotenoid pathway in C.sativus.

The genusCrocusL. (Iridaceae) is monophyletic and contains about 100 species throughout the world.Crocusspecies have horticultural, medicinal and pharmacological importance. Saffron is the dried styles ofC. sativusand is one of the world’s most expensive spices by weight. Controversy exits about the taxonomy of the genus and the species relationship. Exploring genetic diversity and inter-specific cross-ability are important tasks for conservation of wild taxa and for breeding of cultivatedC. sativus. The present study was performed to study genetic variability and population structure in fiveCrocusL. species includingCrocus almehensisBrickell & Mathew,C. caspiusFischer & Meyer,C. speciosusMarschall von Biberstein,C. haussknechtiiBoissier, andC. sativusL. by inter simple sequence repeat (ISSR) molecular markers. We also used published internal transcribed spacer (ITS) sequences to study species relationship and compare the results with ISSR data. The results revealed a high degree of genetic variability both within and among the studied species. Neighbor joining (NJ) tree and network analysis revealed that ISSR markers are useful inCrocusspecies delimitation. Population fragmentation occurred inC. caspiusandC. sativus. Both ISSR and sequenced based analyses separatedC. sativusfrom the other studied species. Close genetic affinity ofC. sativusandC. pallisiiand inter-specific gene flow was supported by both data sets.

[EN] Saffron, used in cookery as a flavouring and colouring agent, is well-known for its antioxidant and beneficial health properties. In the present work, the effect of saffron addition (0-control, 0.1, 0.2 and 0.4%, w/w) in the formulation of fresh pasta was evaluated on textural, physical-chemical, and sensory properties of the cooked product. Content and retention of the bioactive molecules of saffron (crocins) were evaluated by HPLC along with the corresponding antioxidant activity of enriched pasta. The presence of saffron significantly influenced textural and physical-chemical properties of pasta. Higher saffron concentrations enhanced the antioxidant activity of pasta with the higher values of crocins in samples enriched with 0.4% saffron extract even after 3 min of cooking (4.23-5.06 mg/g db). Sensory analysis showed an increased acceptability of the saffron enriched pasta for all descriptors selected (visual aspect, colour, aroma, taste, chewiness, hardness, gumminess and overall acceptability)., Authors are thankful to the Province of Bolzano for financial support (Landesregierung mittels Beschluss Nr. 1472, 07.10.2013) and also the Research Institute for Food Research and Development at the Universitat Politecnica de Valencia (UPV), for laboratory facilities.

Native Crocus taxa are abundant in regions with moderate and dry climate ; however, their potential in the food industry has thus far been neglected. The objective of this study was to compare the quantitative and qualitative flavonoid content and antioxidant and cytotoxic activity in organs of several populations of Croatian native taxa and assess their potential for application in the food industry. The dominant flavonoids in the native Crocus taxa were kaempferol and quercetin. Tepals contained similar or higher concentrations of flavonoid- glycosides than other organs. Tepals from Cvv1 and Cvv2 populations contained more flavonoid- glycosides than the commonly used spice saffron (C. sativus stigmas). The FRAP antioxidant activity of Cvv1 and Cvv5 tepals was similar to that of standard Trolox. DPPH inhibition of Cvv1 and Cvv3 tepals was within the range of that recorded for saffron. Cvv1 tepals significantly reduced reactive oxygen species in the broadest concentration range (50–1000 μg/ml), and showed considerable antioxidant activity in the ABTS assay, equal to 82% of standard Trolox antioxidant activity. A significantly higher concentration of kaempferol-rutinoside was recorded in this than in other taxa. The flavonoid showed a very strong or strong correlation with antioxidant assays results, and a negative correlation with cellular reactive oxygen species concentration. We therefore presumed that kaempferol- rutinoside is one of the main antioxidant phenolics in Crocus tepals. None of the tested extracts showed cytotoxicity toward Caco-2 cells. The results revealed that Cvv tepals have potential as a food supplement and are a promising material for further food safety tests.

Safran, Crocus sativus, gilt weltweit als das teuerste Gewürz. Um die Evolution vom triploidem Safran nachzuvollziehen wählte ich hier einen hierarchischen Ansatz von der Analyse der Phylogenie der Serie zu der C. sativus gehört, über die Studie potentieller Eltern und ihrer geographischen Vorkommen, hin zur Identifikation wichtiger Gene des Sekundärstoffwechsels und ihr Vergleich zwischen Wild- und Kulturform des Safran. Wir konnten die verwandtschaftlichen Beziehungen Safrans innerhalb der Serie Crocus klären und den diploiden C. cartwrightianus, welcher auch als "wilder Safran" bezeichnet wird, als Vorfahren identifizieren. Dieser war an der Bildung und Domestizierung von Safran im südlichen Festland Griechenlands involviert war. Obwohl wir kein C. cartwrightianus Individuum mit völlig identischen Allelmuster im Vergleich zu C. sativus finden konnten, zeigte sich, dass einige C. cartwrightianus Individuen aus dem Süden Attikas ähnliche Expressionsmuster von mutmaßlich an der Carotinoidbiosynthese beteiligten Genen aufwiese wie Safran., Saffron, Crocus sativus, is known as the most expensive spice worldwide. To trace the evolution of triploid saffron, we performed a nested analysis approach reaching from phylogeny of the species group where saffron belongs to, through detection of the parents or closest relatives and identification of the area of domestication of the crop, to the analysis of genes involved in metabolite synthesis of important apocarotenoids. We clarified the relationships of the crop within series Crocus and could identify diploid C. cartwrightianus, sometimes called ‘wild saffron,’ as the progenitor involved in the formation and domestication of saffron in southern mainland Greece. Although, we could not find a C. cartwrightianus individual with the completely identical allele pattern of C. sativus, we found some C. cartwrightianus individuals from southern Attica showing more similar gene expression patterns to saffron in terms of putative genes involved in the carotenoid biosynthesis of saffron.

This review focuses on in vitro culture of saffron ( Crocus sativus L.) based on considerable researches for the last two decades. Multiple endeavors have been taken to modernize the cultivation of the saffron to fulfill the requirement efficient large-scale production of pathogen-free corms, for which tissue culture was proven ideal. The usage of tissue culture techniques has been successfully implemented with the saffron. Here in this review special emphasis has been given to influence of explant and carbohydrate source, plant hormones, and microenvironment during in vitro culture. The cumulative information from this review would be of indispensible importance for the Crocus scientists for further exploration aiming quality enhancement of this potential herb.

This study is part of a wider project on the bitter taste of saffron and its preparations. A deeper knowledge on the taste perception of picrocrocin is necessary in order to develop products that satisfy consumer senses and provide them with adequate amounts of saffron major constituents, also appreciated for bioactivity. A systematic approach on the bitterness of picrocrocin, the major responsible compound, was conducted. A panel was trained specifically for the determination of taste detection and recognition thresholds of picrocrocin, which were found to be 5.34 and 7.26 mg/L, respectively, using the Ascending Forced Choice of Limits methodology. The threshold values were examined in water in absence and presence of other saffron constituents and ethanol and were found to decrease when served hot (61 ± 4 °C). Bitterness was enhanced in 40% (v/v) aqueous ethanol. In both aqueous and ethanol extracts, the presence of saffron volatiles improved bitterness perception. The usefulness of the study was tested in the case of commercial saffron based infusions.

Crocetin is one of the major active ingredients of the rare medicinal herb saffron (Crocus sativus L.), also known as crocus. Crocetin has a wide range of effects including antitumor, anticancer, antioxidant, and antiatherosclerotic activity. Crocetin is currently only obtained via extraction from crocus stigmas, with limited yields. This study developed a new method of production of crocetin from genetically engineered Chlorella vulgaris, a unicellular, fast-growing, and heterotrophically cultured green microalga. A plant expression vector carrying the genes crtRB and ZCD1, which encode enzymes controlling critical steps in crocetin biosynthesis, was introduced into C. vulgaris using the Agrobacterium tumefaciens-mediated transformation method. After hygromycin screening, resistant C. vulgaris strains were achieved. Polymerase chain reaction amplification confirmed that the crtRB and ZCD1 were successfully integrated into the C. vulgaris genome. High-performance liquid chromatography analysis of methanol extraction showed that the transgenic C. vulgaris was able to produce crocetin, while wild-type C. vulgaris was not. These results strongly suggest that introducing crtRB and ZCD1 into C. vulgaris can successfully produce crocetin. This is a new method of crocetin production in C. vulgaris to compensate for the shortage of crocetin.

Saffron, obtained from the flower stigmas of Crocus sativusL., is one of the most expensive food spices. The introduction of saffron in alpine areas could help to broaden and diversify the activities of mountain multifunctional farms, with a positive impact on economy and land management. According to ISO 3632 (2010/2011), saffron can be classified into three categories of quality (I, II, III) depending on the concentration of the three main metabolites responsible for its characteristic colour, flavor and aroma: Crocin, Picrocrocin and Safranal. This study represents the first investigation of the quality of saffron produced in the Italian Alps evaluated with spectrophotometry, HPLC,solid-phase microextraction (SPME), and gas chromatographic analysis combined with mass spectrometry (GC/MS). The experiments used Crocus sativusstigmas produced in 2012-2013 in different areas of the Central Italian Alps were located at an altitude between 720 and 1200 m a.s.l.. Results obtained were compared to commercial saffron. The analyses confirmed that all samples can be classified in the first quality category according to the ISO classification. This high quality is also confirmed by HPLC analysis. Moreover, the SPME-GC/MS analysis identified some differences in the aromatic profile of saffron samples, in particular regarding safranal concentration. A preliminary assessment of the economic viability of high quality saffron production for local markets was also performed. Our study provides valid information regarding the quality and economic sustainability of saffron production in the alpine area confirming this crop as a good candidate for a new source of income for multifunctional farms in mountain areas.

The cellular transport and bioactivity of the second major saffron apocarotenoid, picrocrocin, was examined in parallel to that of the major group, crocetin sugar esters, in aqueous extracts. The transport of pure picrocrocin was investigated in comparison to that of other saffron apocarotenoids, trans-crocetin (di-β-D-gentiobiosyl) ester and crocetin using the Caco-2 cell model coupled with an in vitro digestion procedure. RP-HPLC-DAD was employed to quantify the bioaccessible and bioavailable amounts of individual apocarotenoids. Picrocrocin and crocetin sugar esters though highly bioaccessible (75% and 60%, respectively) were transported at minute quantities (0.2% and 0.5%, respectively; 10-fold lower than crocetin). Picrocrocin did not protect against oxidant-induced DNA damage in U937, human monocytic blood cells at the concentration investigated, however, it reduced the proliferation of human adenocarcinoma and hepatocarcinoma cells. Our findings may be useful for the requirements of food legislation regarding saffron preparations, in which both apocarotenoid groups coexist.

The apocarotenoid crocetin and its glycosylated derivatives, crocins, confer the red colour to saffron. Crocetin biosynthesis in saffron is catalysed by the carotenoid cleavage dioxygenase CCD2 (AIG94929). No homologues have been identified in other plant species due to the very limited presence of crocetin and its derivatives in the plant kingdom. Spring Crocus species with yellow flowers accumulate crocins in the stigma and tepals. Four carotenoid CCDs, namely CaCCD1, CaCCD2 and CaCCD4a/b and CaCCD4c were first cloned and characterized. CaCCD2 was localized in plastids, and a longer CCD2 version, CsCCD2L, was also localized in this compartment. The activity of CaCCD2 was assessed in Escherichia coli and in a stable rice gene function characterization system, demonstrating the production of crocetin in both systems. The expression of all isolated CCDs was evaluated in stigma and tepals at three key developmental stages in relation with apocarotenoid accumulation. CaCCD2 expression parallels crocin accumulation, but C14 apocarotenoids most likely are associated to the CaCCD1 activity in Crocus ancyrensis flowers. The specific CCD2 localization and its membrane interaction will contribute to the development of a better understanding of the mechanism of crocetin biosynthesis and regulation in the chromoplast.

Mediterranean characteristics are attributed to the genusCrocus, which is inadequately studied in terms of seed germination. An afterripening requirement is very common in environments with warm and dry periods, and photoinhibition has been detected in many angiosperms inhabiting dry and open areas. The effects of afterripening and light on seed germination were investigated for the first time in 23 native GreekCrocustaxa, collected from various localities with either a Mediterranean or a temperate climate. Germination experiments were conducted in continuous darkness and in light at the optimal temperature for each taxon, with both freshly collected and afterripened seeds; warm stratification (20°C, darkness) was also examined in 22 taxa. A number of selected taxa were additionally investigated with respect to afterripening outdoors, afterripening and warm stratification at higher temperatures (35 and 25°C, respectively), stratification at 20/10°C, dry storage at low temperatures, response to gibberellic acid and phenology of embryo growth. It was postulated that an afterripening requirement is a characteristic of the genusCrocus, and we found that it can be fulfilled in nature during the Mediterranean dry summer. Also, for the vast majority of the taxa, warm stratification and stratification at 20/10°C can both meet the afterripening requirement. Embryos of the taxa studied are underdeveloped and have to grow prior to germination. Intrageneric differences of seed germination were observed only towards light, with photoinhibition being predominant in taxa from drier environments.

// Parthasarathy Rangarajan 2 , Dharmalingam Subramaniam 2 , Santanu Paul 1 , Deep Kwatra 2 , Kanagaraj Palaniyandi 1 , Shamima Islam 1 , Sitaram Harihar 1 , Satish Ramalingam 2 , William Gutheil 3 , Sandeep Putty 3 , Rohan Pradhan 4 , Subhash Padhye 4 , Danny R. Welch 1, 2 , Shrikant Anant 1, 2 , Animesh Dhar 1, 2 1 Department of Cancer Biology, University of Kansas Medical Center, Kansas City, KS, USA 2 Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA 3 Department of Pharmaceutical Sciences, University of Missouri at Kansas City, Kansas City, MO, USA 4 Interdisciplinary Science and Technology Research Academy, Abeda Inamdar College, University of Pune, Pune, India Correspondence to: Animesh Dhar, e-mail: adhar@kumc.edu Keywords: crocetin, nude mice, pancosphere, DCLK-1, gli transcription factor Received: April 02, 2015 Accepted: July 31, 2015 Published: August 13, 2015 ABSTRACT Pancreatic cancer is the fourth leading cause of cancer deaths in the US and no significant treatment is currently available. Here, we describe the effect of crocetinic acid, which we purified from commercial saffron compound crocetin using high performance liquid chromatography. Crocetinic acid inhibits proliferation of pancreatic cancer cell lines in a dose- and time-dependent manner. In addition, it induced apoptosis. Moreover, the compound significantly inhibited epidermal growth factor receptor and Akt phosphorylation. Furthermore, crocetinic acid decreased the number and size of the pancospheres in a dose-dependent manner, and suppressed the expression of the marker protein DCLK-1 (Doublecortin Calcium/Calmodulin-Dependent Kinase-1) suggesting that crocetinic acid targets cancer stem cells (CSC). To understand the mechanism of CSC inhibition, the signaling pathways affected by purified crocetinic acid were dissected. Sonic hedgehog (Shh) upon binding to its cognate receptor patched, allows smoothened to accumulate and activate Gli transcription factor. Crocetinic acid inhibited the expression of both Shh and smoothened. Finally, these data were confirmed in vivo where the compound at a dose of 0.5 mg/Kg bw suppressed growth of tumor xenografts. Collectively, these data suggest that purified crocetinic acid inhibits pancreatic CSC, thereby inhibiting pancreatic tumorigenesis.

Background and Aims: Saffron (Crocus sativus) is a sterile triploid (2n = 3x = 24) cultivated species, of unknown origin from other diploid and polyploid species in the genus Crocus (Iridaceae). Species in the genus have high morphological diversity, with no clear phylogenetic patterns below the level of section Crocus series Crocus. Using DNA markers, this study aimed to examine the diversity and relationships within and between species of Crocus series Crocus. Methods: Eleven inter-retroelement amplified polymorphism (IRAP) primers were used in 63 different combinations with 35 single-plant accessions of C. sativus and related Crocus species in order to determine genetic variability and to conduct phylogenetic analysis. Key Results: A total of 4521 distinct polymorphic bands from 100 bp to approx. 4 kb were amplified; no fragment specific to all accessions of a single species was amplified. The polymorphic information content (PIC) values varied from approx. 0·37 to approx. 0·05 (mean 0·17 ± 0·1) and the major allele frequency had a mean of 0·87. High levels of polymorphism were identified between accessions of the six species of Crocus series Crocus related to C. sativus, with further variation between the species. In contrast, no polymorphisms were seen among 17 C. sativus accessions obtained in the region from Kashmir through Iran to Spain. Conclusions In contrast to the intraspecific variability seen in other Crocus species, C. sativus has minimal genetic variation, and it is concluded that the triploid hybrid species has most probably arisen only once. The data show that saffron is an allotriploid species, with the IRAP analysis indicating that the most likely ancestors are C. cartwrightianus and C. pallasii subsp. pallasii (or close relatives). The results may facilitate resynthesizing saffron with improved characteristics, and show the need for conservation and collection of wild Crocus.

All retinal disorders, regardless of their aetiology, involve the activation of oxidative stress and apoptosis pathways. The administration of neuroprotective factors is crucial in all phases of the pathology, even when vision has been completely lost. The retina is one of the most susceptible tissues to reactive oxygen species damage. On the other hand, proper development and functioning of the retina requires a precise balance between the processes of proliferation, differentiation and programmed cell death. The life-or-death decision seems to be the result of a complex balance between pro- and anti-apoptotic signals. It has been recently shown the efficacy of natural products to slow retinal degenerative process through different pathways. In this review, we assess the neuroprotective effect of two compounds used in the ancient pharmacopoeia. On one hand, it has been demonstrated that administration of the saffron constituent safranal to P23H rats, an animal model of retinitis pigmentosa, preserves photoreceptor morphology and number, the capillary network and the visual response. On the other hand, it has been shown that systemic administration of tauroursodeoxycholic acid (TUDCA), the major component of bear bile, to P23H rats preserves cone and rod structure and function, together with their contact with postsynaptic neurons. The neuroprotective effects of safranal and TUDCA make these compounds potentially useful for therapeutic applications in retinal degenerative diseases. This research was supported by grants from the Spanish Ministry of Economy and Competitiveness-FEDER (BFU2012-36845), Instituto de Salud Carlos III (RETICS RD12/0034/0010), Asociación Retina Asturias, Fundación Jesús Gangoiti, Organización Nacional de Ciegos Españoles (ONCE) and FUNDALUCE.

Crocus sativus L. belongs to family Iridaceae and its stigma part which forms commercial saffron has been used as a spice and medicine ever since the Greek-Minoan civilization. Saffron is considered the most expensive spice in the world and the site for the biosynthesis of peculiar metabolites (apocarotenoids), responsible for its characteristic color, flavor, and aroma. There is enough literature citing the antioxidant properties of saffron metabolites, but most of the knowledge accumulated so far comes from stigma part only, while other plant parts particularly the vegetative tissues have not been worked out. The present study was conducted to evaluate and compare the chemical composition and antioxidant activity of three different tissue types of Crocus viz: stigma, corm, and leaf. The phytochemical analysis carried out using LC-MS showed that the major constituents identified were flavonoids like kaempferol, taxifolin, naringenin, etc. and apocarotenoids including crocin, crocetin, and their derivatives. Also the total phenolic, flavonoid, and carotenoid contents were determined. The antioxidant property of these tissue types was also investigated and compared by biochemical assays like, DPPH, NBT, and FRAP. The ethanolic fraction of stigma demonstrated the strongest antioxidant activity which could be attributed to its highest content of phenolics and flavonoids. Stigma extract was further evaluated for its role in alleviating oxidative stress in plants, yeast, and bacteria. The results showed that stigma extract reduced methylviologen induced chlorophyll damage, lipid peroxidation, and protein oxidation in plants thereby rendering them more tolerant to stress. It was also shown to alleviate H2O2 mediated oxidative stress tolerance in bacteria and yeast.

Population growth and world climate changes are putting high pressure on agri-food production systems. Exacerbating use of energy sources and expanding the environmental damaging symptoms are the results of these difficult situations. This study was conducted to determine the energy balance for saffron production cycle and investigate the corresponding greenhouse gas (GHG) emissions in Iran. Saffron (Crocus sativus L.) is one of the main spice that historically cultivated in Iran. Data were obtained from 127 randomly selected saffron growers using a face to face questionnaire technique. The results revealed that in 5 years of saffron production cycle, the overall input and output energy use were to be 163,912.09 and 184,868.28 MJ ha(-1), respectively. The highest-level of energy consumption belongs to seeds (23.7 %) followed by chemical fertilizers (23.4 %). Energy use efficiency, specific energy, net energy, and energy productivity of saffron production were 1.1, 13.4 MJ kg(-1), 20,956.2 MJ ha(-1), and 0.1 kg MJ(-1), respectively. The result shows that the cultivation of saffron emits 2325.5 kg CO2 eq. ha(-1) greenhouse gas, in which around 46.5 % belonged to electricity followed by chemical fertilizers. In addition the Cobb-Douglas production function was applied into EViews 7 software to define the functional relationship. The results of econometric model estimation showed that the impact of human labor, electricity, and water for irrigation on stigma, human labor, electricity, and seed on corm and also human labor and farmyard manure (FYM) on flower and leaf yield were found to be statistically significant. Sensitivity analysis results of the energy inputs demonstrated that the marginal physical productivity (MPP) worth of electricity energy was the highest for saffron stigma and corm, although saffron flower and leaf had more sensitivity on chemicals energy inputs. Moreover, MPP values of renewable and indirect energies were higher than non-renewable and direct energies, respectively.

We present a new extraction protocol, using ethyl alcohol as a solvent, to evaluate safranal by gas chromatography (GC). A linear response was obtained with R(2)=0.995 and a reproducibility standard deviation of 4.7-6.0%. The limit of detection and limit of quantitation were 0.05 and 0.25gkg(-1), respectively. The GC data for several samples of powdered saffron from different origins were compared to specific absorbance values measured according to the ISO Normative 3632-1:2011 method. The aroma strength of saffron samples quantitated by GC and the specific absorbance values of safranal by the UV method did not correlate. Quantitative evaluation of safranal by GC appears to be more specific and useful for commercial comparisons of saffron quality.

The flora of Jordan with regards to the flowering vascular plant represents a high diversity in such a semi-arid country like Jordan which is part of the Eastern Mediterranean region. In this study, a number of 59 wild species with showy, exotic flowers from Jordan are illustrated; the selected species are not introduced or cultivated but rather local wild flowers selected for their showy, colorful flowers and for their medicinal, edible and ornamental values. Most of the recorded species in this study are endangered and exposed to degradation. The selected plant species are ornamental, showy with colorful flowers like Orchids, Irises, Crocuses, Narcissus, Cyclamen, Ixiolirion, Linum, Anemone, Salvias, Sternbergia and Ornithogallum. Some of the illustrated plant species are endemic to Jordan like: Iris nigricans which is the national flower of Jordan, Crocus moabiticus, Iris edomensis and Iris petrana, many plant species are demonstrated for their medicinal values like Achillea fragrantissima, Arum palestinum, Glaucium arabicum, Erodium gruinum, Alcea setosa, Hyoscyamus reticulatus and others. Other species are mentioned for their edible consumption like Capparis spinosa, Cretaegus aronia, Crocus hermoneus, Cyclamen persicum, Gundelia tournefortii, Lupinus varius, Malva setosa and others. Photographs are demonstrated for most of the selected plants. Key words: Exotic flowers, wild flowers, Jordan.

A chromatographic method was developed and fully validated for the determination of the major saffron constituents, i.e., picrocrocin and five major crocins. Dried samples (styles of Crocus sativus and other Crocus taxa) were extracted with MeOH : water (1 : 1, v/v), and chromatographic separation of the analytes was achieved by reversed-phase chromatography using a gradient elution. Full validation was performed using spiked samples with analytes, which were isolated, purified, and characterized by MS due to a lack of commercial standards. The method showed a good fit (r2 0.999) for all analytes with limit of quantitation values in the range of 1-15 µg/mL, and demonstrated adequate intra- and inter-precision ( 15 % RSD) and accuracy ( 7 % RE). The method was applied to the analysis of various commercial saffron samples and of indigenous Crocus taxa and allowed for the first time the absolute quantitation of several Crocus components.