Antimicrobial activity, synergism and inhibition of germ tube formation by Crocus sativus-derived compounds against Candida spp

The limited arsenal of synthetic antifungal agents and the emergence of resistant Candida strains have prompted the researchers towards the investigation of naturally occurring compounds or their semisynthetic derivatives in order to propose new innovative hit compounds or new antifungal combinations endowed with reduced toxicity. We explored the anti-Candida effects, for the first time, of two bioactive compounds from Crocus sativus stigmas, namely crocin 1 and safranal, and some semisynthetic derivatives of safranal obtaining promising biological results in terms of minimum inhibitory concentration/minimum fungicidal concentration (MIC/MFC) values, synergism and reduction in the germ tube formation. Safranal and its thiosemicarbazone derivative 5 were shown to display good activity against Candida spp

A study of the role of nitric oxide in the mechanism of action of hydroalcoholic extract of saffron (Crocus sativus) on the electrophysiological properties of the rabbit atrioventricular node

Biologically active substances of plant origin represent an essential branch of modern cardiovascular pharmacotherapy. Furthermore, drugs of plant origin have the advantage of weaker adverse effects and lower prices than synthetic drugs. Pharmacological studies and traditional medical literature point to the anti-ischemic and hypotensive effects of the Crocus sativus L. (Iridacea). The major goals of the present study were: (1) to determine the negative dormotrophic properties of a hydroalcoholic extract of saffron on an isolated AV node and (2) to establish the role of nitric oxide in the mediating effects of saffron on the electrophysiological properties of the AV node. This was an experimental study. Selective stimulation protocols were used to independently quantify AV nodal recovery, facilitation and fatigue. We used isolated perfused rabbit AV node preparation, in three groups (N=32); in each group, we assessed the plant's effect in comparison with the control. In the pilot study, we used different concentrations (A=9 x 10-2 mg/L, B=19 x 10-2 mg/L and C=27 x 10-2 mg/L) to select the optimum concentration (19 x 10-2 mg/L) of the hydroalcoholic extract of saffron. Saffron has a depressant effect on basic and rate-dependent properties of the AV node. We observed an increasing AVCT (38.8 ±4 to 41.7 ±4 msec) and FRP (157.6 ±3 to 163.7 ±4 msec). Also saffron increased the amount of facilitation and the magnitude of fatigue (5.9 ±0.3 to 11.1 ±1 msec). The NOS inhibitor (L-NAME) has a preventative effect on the depressant effect of saffron on AVCT and FRP

Physicochemical Characterization of Safranal-β-Cyclodextrin Inclusion Complexes Prepared By Supercritical Carbon Dioxide and Conventional Methods.

Saffron (Crocus sativus. Linn) has attracted much attention over the last decade because it has a large number of potent and biologically active compounds such as crocin, crocetin, picrocrocin and safranal. Researchers have shown that safranal has high antioxidant and cytotoxicity activities against several types of tumour cells (e.g., hepatocellular carcinoma) both in-vitro and in-vivo. However, the low aqueous solubility of safranal prevents using it as a therapeutic or preventive agent. β-cyclodextrin (β-CD) inclusion complexes are being used in pharmaceutical applications to alter physicochemical properties (e.g., solubility, volatility, stability, chemical reactivity, and bio-availability) of poorly water soluble drugs. Thus the aim of this work is to investigate the potential of different methods for the preparation of safranal-β-CD inclusion complexes in order to enhance the poor solubility and dissolution rate of safranal in aqueous solutions. Inclusion complexes having a molar ratio of 1:2 (safranal-β-CD) were prepared using different methods such as kneading (KN), co-evaporation (COE), sealed-heating (SH), and supercritical CO2 (SC-CO2). Fourier transform infrared spectroscopy (FT-IR), X-ray powder diffraction (XRD), scanning electron microscopy (SEM) and proton nuclear magnetic resonance spectroscopy (H-NMR) were used to identify the physicochemical properties of inclusion complexes. Phase solubility and dissolution measurements were also studied. Effects of temperature (35 and 55 °C) and pressure (100 and 300 bar) on the inclusion complexes prepared using the SC-CO2 method were also investigated. The results indicated that the formation of safranal-β-CD inclusion complexes was affected by the preparation method being used, and the SC-CO2 method proved to be more effective than conventional techniques. FT-IR and H-NMR results indicated the formation of inclusion between β-CD and safranal in the complex prepared by different methods. A “Bs” type solubility with an apparent solubility constant (Ks) of 51.48 M-1 for safranal was obtained from the initial slope of phase solubility diagram. The intrinsic solubility of safranal was increased from 3.852 mM to 5.217 mM in the presence of 10.00 mM of β-CD. Therefore, the initial phase solubility of safranal was enhanced by about 35% in water solution. Dissolution rate studies showed that inclusion complexes might dissolve faster than pure safranal or even physical mixture of safranal-β-CD. Hence, β-CD could be useful for solid safranal formulations. The solvent-free product prepared by SC-CO2 showed high aqueous solubility and may provide minimal side effects for human use

Crocin loaded nano-emulsions: Factors affecting emulsion properties in spontaneous emulsification

Spontaneous emulsification may be used for encapsulating bioactive compounds in food and pharmaceutical industry. It has several advantages over high energy and other low energy methods including, protecting sensitive compounds against severe conditions of high energy method and its ability to minimize surfactant, removal of cosurfactant and thermal stability compared with other low energy methods. In this study, we examined possibility of encapsulating highly soluble crocin in W/O micro-emulsions using spontaneous method which further could be used for making double emulsions. Nonionic surfactants of Span 80 and polyglycerol polyricinoleate (PGPR) were used for making micro-emulsions that showed the high potential of PGPR for spontaneous method. Surfactant to water ratio (SWR) was evaluated to find the highest amount of aqueous phase which can be dispersed in organic phase. Droplet size decreased by increasing SWR toward the SWR = 100 which had the smallest droplet size and then increased at higher levels of surfactant. By increasing SWR, shear viscosity increased which showed the high effect of PGPR on rheological properties. This study shows in addition to W/O micro-emulsions, spontaneous method could be used for preparing stable O/W micro-emulsions. © 2015 Elsevier B.V

Crocus Sativus L. (Saffron) Extract and its Active Constituents (Crocin and Safranal) on Ischemia-Reperfusion in Rat Skeletal Muscle

Saffron and its constituents have been shown to decrease ischemia-reperfusion (I/R) injury in kidney or brain tissues. In this study, the effects of saffron ethanolic extract and its constituents, crocin and safranal, were evaluated in skeletal muscle during I/R injury. Hind limb ischemia was induced using clamping the common femoral artery and vein. After 2 h ischemia, the clamp of the femoral vessels of animals was taken off and the animal underwent 1h reperfusion. Muscle injuries were evaluated by recording of the electromyographic (EMG) potentials and performing some biochemical analysis including thiobarbituric acid reactive substances (TBARS), total sulfhydryl (SH) groups and antioxidant capacity of muscle (using FRAP assay). The ethanolic extract of saffron (5, 20 and 80 mg kg−1), crocin (50, 200 and 400 mg kg−1), safranal (0.1, 0.25 and 0.5 ml kg−1) and normal saline (10 ml kg−1) were administered intraperitoneally 1 h prior reperfusion. The average peak-to-peak amplitude during I/R was significantly increased in extract, crocin and safranal groups in comparison with control-ischemic group. Following saffron, crocin and safranal administration, the total SH contents and antioxidant capacity were elevated in muscle flap. The MDA level was declined significantly in test groups. It is concluded that saffron extract and its constituents show a protective effect against lower limb I/R in rat

Saffron and its major ingredients’ effect on colon cancer cells with mismatch repair deficiency and microsatellite instability

Background: Colorectal cancer (CRC) is one of the most common cancers worldwide. One of its subtypes is associated with defective mismatch repair (dMMR) genes. Saffron has many potentially protective roles against colon malignancy. However, these roles in the context of dMMR tumors have not been explored. In this study, we aimed to investigate the effects of saffron and its constituents in CRC cell lines with dMMR. Methods: Saffron crude extracts and specific compounds (safranal and crocin) were used in the human colorectal cancer cell lines HCT116, HCT116+3 (inserted MLH1), HCT116+5 (inserted MSH3), and HCT116+3+5 (inserted MLH1 and MSH3). CDC25b, p-H2AX, TPDP1, and GAPDH were analyzed by Western blot. Proliferation and cytotoxicity were analyzed by MTT. The scratch wound assay was also performed. Results: Saffron crude extracts restricted (up to 70%) the proliferation in colon cells with deficient MMR (HCT116) compared to proficient MMR. The wound healing assay indicates that deficient MMR cells are doing better (up to 90%) than proficient MMR cells when treated with saffron. CDC25b and TDP1 downregulated (up to 20-fold) in proficient MMR cells compared to deficient MMR cells, while p.H2AX was significantly upregulated in both cell types, particularly at >10 mg/mL saffron in a concentration-dependent manner. The reduction in cellular proliferation was accompanied with upregulation of caspase 3 and 7. The major active saffron compounds, safranal and crocin reproduced most of the saffron crude extracts' effects. Conclusions: Saffron's anti-proliferative effect is significant in cells with deficient MMR. This novel effect may have therapeutic implications and benefits for MSI CRC patients who are generally not recommended for the 5-fluorouracil-based treatment

SIMULTANEOUS EXTRACTION AND RAPID HPLC BASED QUANTIFICATION OF CROCIN AND SAFRANAL IN SAFFRON (CROCUS SATIVUS L.)

Objective: Saffron (Crocus sativus L.) is one of the most valuable crops with high medicinal values used in many diseases. The aim of the study was to establish the reliability and rapid HPLC method for analysis of crocin and safranal concentration present in stigmata of saffron in the market as well as field grown samples.Methods: Field grown and market stigmata of saffron were used for extraction of crocin and safranal. The linear dynamic ranges were established after validating the robustness of critical method parameters. The Agilent1260-Infinity Quaternary LC system was used for the preparation of calibration standards and quantification of crocin and safranal in C. sativus stigmata.Results: A good linearity was achieved in the range of 10-30 µg for each compound with the determination coefficient (R2). The calibration curves revealed linear regression (r. 0.997) for this rapid HPLC method and limit of quantifications (LOQs) were achieved in the range of 3.4 µg/ml for crocin and 10.2 µg/ml for safranal. The limit of detection (LODs) for all standards was ≤4.2 µg/ml. The range of crocin content (10.43-16.32 mg/g) and safranal (5.19-5.21 mg/g) was estimated in saffron samples.Conclusion: This method may serve the purpose of accurate quantification of crocin and safranal present in the stigmata of the plant in a quick time period.Keywords: Saffron, Crocin, Safranal, Extraction, Quantification, HPL

Distribution of Main Bioactive Compounds from Saffron Species as a Function of Infusion Temperature and Time in an Oil/Water System

Most research on saffron has focused on its composition and beneficial effects, while the culinary perspective to enhance its gastronomic potential remains unexplored. This study aims to define the transfer of the main compounds responsible for color, flavor, and aromatic properties, evaluating three critical variables: temperature (60 °C, 80 °C and 100 °C), infusion time (ranging from 10 to 30 min), and the composition of the medium (water, oil, and water/oil). Samples were analyzed using the LC-QTOF MS/MS and ISO 3632-1:2011 methods. The major compounds were crocins, including trans-crocin and picrocrocin. Among the flavonoids, kaempferol 3-O-sophoroside stands out. Regarding extraction conditions, crocins, glycoside flavonoids, and picrocrocin were enhanced in water, the former in 100% water and at low temperatures, while picrocrocin proved to be the most stable compound with extraction favored at high temperatures. The variable with the greatest incidence of picrocrocin isolation seemed to be the concentration of water since water/oil compositions reported higher concentrations. Safranal and kaempferol were enriched in the oil phase and at lower temperatures. This study provides a chemical interpretation for the appropriate gastronomic use of saffron according to its versatility. Finally, the determination of safranal using the ISO method did not correlate with that obtained using chromatography

The synthesis of safranal

Thesis (B.S.)--Massachusetts Institute of Technology, Dept. of Chemistry, 1953.MIT copy bound with: Solution and emulsion copolymerization of alkenyl siloxanes with vinyl type monomers / by Earl W. Mitchell.Includes bibliographical references (leaf 18).Introduction: Safranal is a cyclic monoterpene which occurs naturally in the saffron plant as a glucoside, picrocrocin. In 1922 Winterstein and TeleCzky obtained safranal by hydrolysis of picrocrocin, but its structure was not determined. Kuhn and Winterstein hydrolyzed picrocrocin in 1933 and obtained safranal and glucose in a one to one ratio. The structures for safranal and picrocrocin were determined. [illustration] Since this time there have been many attempts to devise a practical synthesis of safranal. Among these has been the work of Kuhn and Wendt, who obtained safranal in 1-3% yield by dehydrogenation of [beta]-cyclocitral. Karrer and Ochsner attempted its synthesis by bromination of [alpha]-cyclocitral with N-bromosuccinimide followed by dehydrobromination, but a rearrangement took place. Lunt and Sondheimer obtained a homolog of safranal, 4-methyl safranal, by a Diels-Alder type condensation. Other attempted syntheses have proven unsuccessful. This paper presents the results of attempts to synthesize safranal from the enol acetate of cyclocitral. It is a continuation of the work begun by Vittimberg in 1951. He succeeded in obtaining the enol acetate of cn good yield and determined its structure. Cyclocitral was prepared by first forming a Schiff base from citral (I) and aniline. This was cyclized with concentrated sulfuric acid and steam-distilled to give a mixture of [alpha]- and [beta]-cyclocitral (II and III) in a ratio of 2/3 to 1/3. This mixture was treated with a large excess of isopropenyl acetate to form the enol acetate of cyclocitral. Both [alpha]- and [beta]-cyclocitral give the same enol acetate (IV).by David A. Nelson.B.S