crocin and picrocrocin

Crocus sativus is an herbaceous plant that belongs to family Iridaceae. It is commonly known as saffron and has been used for medicinal purposes since many centuries in India and other parts of the world. Saffron of commercial importance comprises of dried stigmas of the plant and is rich in flavonoids, vitamins, and carotenoids. Carotenoids represent the main components of saffron and their cleavage results in the formation of apocarotenoids such as crocin, picrocrocin, and safranal. Studies conducted during the past two decades have revealed the immense therapeutic potential of saffron. Most of the therapeutic properties are due to the presence of unique apocarotenoids having strong free radical scavenging activity. The mode of action of these apocarotenoids could be: modulatory effects on detoxifying enzymes involved in combating oxidative stress, decreasing telomerase activity, increased the proapoptotic effect, inhibition of DNA, RNA and protein synthesis, and by a strong binding capacity of crocetin with tRNA. The present review focuses on the therapeutic role of saffron and its bio oxidative cleavage products and also highlights the possible molecular mechanism of action. The findings reported in this review describes the wide range of applications of saffron and attributes its free radical scavenging nature the main property which makes this spice a potent chemotherapeutic agent for the treatment of various diseases.

Topics: Animals; Antineoplastic Agents; Carotenoids; Crocus; Cyclohexenes; Flavonoids; Glucosides; Humans; Plant Extracts; Terpenes

Saffron, the dried stigmas of Crocus sativus L., is a highly valued agricultural product that is used mainly as a food coloring and flavoring agent. Three main secondary metabolites of Crocus sativus including crocin, picrocrocin, and safranal are responsible for the color, the bitter taste and for the odor and aroma, respectively. As a component of traditional medicine, saffron has been utilized as a medicinal herb for treating various ailments including cramps, asthma, liver disease, menstruation disorders, pain, and in the pathogenesis of cancer.. To summarize the recent published data on the chemo-preventive properties of Crocus sativus in cancer treatment.. We conducted a non-systematic review of the literature.. A search of English-language literature was performed using Scopus, EMBASE and PubMed. We applied no restriction in time. Articles were searched using the keywords "Lung", "breast", "skin", "prostate", "leukemia", "cancer", "neoplasm", "tumor", "malignancy", "saffron", "crocus sativus", "crocin", "crocetin", "picrocrocin", and "safranal".. Saffron has been reported to exert anti-tumor and anti-cancer effects in various types of cancer including lung cancer, breast cancer, leukemia, skin cancer and prostate cancer. This appears to be via various mechanisms including: the induction of apoptosis, arresting cell cycle progression, suppressing expression of matrix metalloproteinase, modulatory effects on some phase II detoxifying enzymes and decreasing expression of inflammatory molecules are potential mechanisms of saffron-induced anticancer effects.. Saffron possesses potent anti-tumor properties and represents an efficacious and safe treatment.

Topics: Anticarcinogenic Agents; Antineoplastic Agents, Phytogenic; Apoptosis; Carotenoids; Cell Cycle; Crocus; Cyclohexenes; Female; Food Coloring Agents; Glucosides; Humans; Male; Secondary Metabolism; Terpenes; Vitamin A

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.

Topics: Alzheimer Disease; Animals; Atherosclerosis; Carotenoids; Crocus; Cyclohexenes; Depression; Glucosides; Humans; Neoplasms; Phytotherapy; Plant Extracts; Sexual Dysfunction, Physiological; Terpenes; Vitamin A

The spice saffron is made from the dried stigmas of the plant

Topics: Antidepressive Agents; Antineoplastic Agents; Antioxidants; Carotenoids; Color; Crocus; Cyclohexenes; Functional Food; Glucosides; Humans; Phytochemicals; Plants, Medicinal; Spices; Terpenes; Vitamin A

In this work, a four-factor five-level full factorial central composite design (CCD) was used to optimize the ultrasound-assisted extraction (UAE) of saffron major components, namely picrocrocin, safranal and crocin. The process parameters included ethanol concentration (0-100%), extraction time (2-10 min), duty cycle (0.2-1.0) and ultrasonic amplitude (20-100%). The extracted compounds were measured both by spectrophotometry and chromatography techniques. The results revealed that the middle concentrations of ethanol and relatively long process durations along with high duty cycles and ultrasonic amplitudes had the most profound impact on the yields of the extracted bioactives. UAE was optimized using response surface methodology (RSM) and artificial bee colony (ABC); a comparison between these methods indicated their optimization power was approximately the same. According to the RSM analysis, an ethanol concentration of 58.58%, extraction time of 6.85 min, duty cycle of 0.82 and amplitude of 91.11% were the optimum extraction conditions, while the optimal conditions resulting from ABC were 53.07%, 7.32 min, 0.93 and 100% for the UAE variables respectively. Finally, HPLC analysis was carried out on the UAE optimum extract resulting from RSM. Four crocetin esters were detected in the optimal extract.

Topics: Carotenoids; Coloring Agents; Crocus; Cyclohexenes; Ethanol; Glucosides; Plant Extracts; Terpenes

Topics: Carotenoids; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Crocus; Cyclohexenes; Glucosides; Plant Extracts; Tandem Mass Spectrometry; Terpenes

Topics: Carotenoids; Crocus; Dioxygenases; Gardenia; Nicotiana; Zeaxanthins

The dehydration process is a prerequisite to preserve saffron for a long time. According to this process, saffron shows differences in the main compounds responsible for its quality (colour, taste, aroma, and flavonol content). At present, the freeze-drying method obtains dried products with the highest quality. Viruses can modify the physiology and metabolism of plants, being able to affect the activities of several enzymes. For this reason, the main compounds of saffron have been analyzed under two different dehydrating processes, freeze-drying and dark-drying, considering their infection status with the Saffron latent virus (SaLV). Results showed that the picrocrocin and safranal content enables to differ dark-dried samples from freeze-dried ones. Besides, the kaempferol-3-O-sophoroside-7-O-glucoside content allows differentiating between SaLV-infected (SaLV

Topics: Carotenoids; Crocus; Cyclohexenes; Desiccation; Glucosides; Iran; Kaempferols; Phytochemicals; Plant Diseases; Prevalence; Terpenes; Virus Diseases

Saffron stigmas are widely used as food additives and as traditional medicine in Iran and many other countries. The unique taste, flavor and pharmaceutical properties of saffron stigmas are due to the presence of three apocarotenoids secondary metabolites crocin, picrocrocin and safranal. There is limited knowledge about the effect of environmental stresses on the metabolism of apocarotenoids in saffron. We analyzed the content of crocin and picrocrocin and the expression of key genes of apocarotenoid biosynthesis pathways (CsCCD2, CsCCD4, CsUGT2, CsCHY-β and CsLCYB) in saffron plants exposed to moderate (90 mM) and high (150 mM) salt (NaCl) concentrations. Measuring ion concentrations in leaves showed an increased accumulation of Na

Topics: Biosynthetic Pathways; Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Gene Expression Regulation, Plant; Glucosides; Plant Leaves; Potassium; Real-Time Polymerase Chain Reaction; Salt Stress; Sodium; Sodium Chloride; Terpenes

Plant acoustic frequency technology (PAFT) is the effect or treatment of a plant with a specific frequency sound wave.. The sound waves with different frequencies and a sound pressure level 77 dB were emitted on the saffron corms in a controlled environment using aeroponic cultivation and the contents of crocin, picrocrocin and safranal in their produced stigmas were analysed by high-performance liquid chromatography. For this purpose, the corms were divided into two groups. In group 1, sound waves with the frequencies of 0.5, 1 and 2 kHz were emitted on saffron corms in different stages of sprouting, flowering and the whole stage of sprouting and flowering. In group 2, sonication was performed on the corms during the flowering stage at 4, 8, 12 and 16 kHz frequencies.. The changes in the contents of crocin, picrocrocin and safranal were not significantly compared to the control at 0.5, 1 and 2 kHz frequencies in the stages of sprouting and flowering of corms. While the higher frequencies (4, 8, 12 and 16 kHz) in flowering stage were affected significantly, the crocin and picrocrocin content increased 8.5% and 30%, applying the frequency of 12 and 8 kHz, respectively. Also, the effect of sound exposure time per day with the frequency of 16 kHz at 15, 30 and 60 min were investigated.. The findings showed that the corms could be affected by sounding in the different stages of growth of the corm and also in the content of secondary metabolites.

Topics: Carotenoids; Crocus; Cyclohexenes; Glucosides; Plant Extracts; Sound; Terpenes

Topics: Biosynthetic Pathways; Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Gene Expression Regulation, Plant; Glucosides; Nitric Oxide; Salinity; Terpenes

Saffron crocus (Crocus sativus L.) has been widely grown in Iran. Its stigma is considered as the most valuable spice for which several pharmacological activities have been reported in preclinical and clinical studies, the antidepressant effect being the most thoroughly studied and confirmed. This plant part contains several characteristic secondary metabolites, including the carotenoids crocetin and crocin, and the monoterpenoid glucoside picrocrocin, and safranal. Since only the stigma is utilized industrially, huge amount of saffron crocus by-product remains unused. Recently, the number of papers dealing with the chemical and pharmacological analysis of saffron is increasing; however, there are no systematic studies on the chemical variability of the major by-products. In the present study, we harvested saffron crocus flowers from 40 different locations of Iran. The tepals and stamens were separated and subjected to qualitative and quantitative analysis by HPLC-DAD. The presence and amount of seven marker compounds, including crocin, crocetin, picrocrocin, safranal, kaempferol-3-O-sophoroside, kaempferol-3-O-glucoside, and quercetin-3-O-sophoroside were determined. The analytical method was validated for filter compatibility, stability, suitability, accuracy, precision, intermediate precision, and repeatability. Tepal and stamen samples contained three flavonol glycosides. The main constituent of the tepals was kaempferol-3-O-sophoroside (62.19-99.48 mg/g). In the stamen, the amount of flavonoids was lower than in the tepal. The amount of kaempferol-3-O-glucoside, as the most abundant compound, ranged between 1.72-7.44 mg/g. Crocin, crocetin, picrocrocin, and safranal were not detected in any of the analysed samples. Our results point out that saffron crocus by-products, particularly tepals might be considered as rich sources of flavonol glucosides. The data presented here can be useful in setting quality standards for plant parts of C. sativus that are currently considered as by-products of saffron production.

Topics: Antidepressive Agents; Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Flavonoids; Flowers; Glucosides; Kaempferols; Monosaccharides; Plant Extracts; Quercetin; Terpenes; Vitamin A

Crocins and picrocrocin are glycosylated apocarotenoids responsible, respectively, for the color and the unique taste of the saffron spice, known as red gold due to its high price. Several studies have also shown the health-promoting properties of these compounds. However, their high costs hamper the wide use of these metabolites in the pharmaceutical sector. We have developed a virus-driven system to produce remarkable amounts of crocins and picrocrocin in adult Nicotiana benthamiana plants in only two weeks. The system consists of viral clones derived from tobacco etch potyvirus that express specific carotenoid cleavage dioxygenase (CCD) enzymes from Crocus sativus and Buddleja davidii. Metabolic analyses of infected tissues demonstrated that the sole virus-driven expression of C. sativus CsCCD2L or B. davidii BdCCD4.1 resulted in the production of crocins, picrocrocin and safranal. Using the recombinant virus that expressed CsCCD2L, accumulations of 0.2% of crocins and 0.8% of picrocrocin in leaf dry weight were reached in only two weeks. In an attempt to improve apocarotenoid content in N. benthamiana, co-expression of CsCCD2L with other carotenogenic enzymes, such as Pantoea ananatis phytoene synthase (PaCrtB) and saffron β-carotene hydroxylase 2 (BCH2), was performed using the same viral system. This combinatorial approach led to an additional crocin increase up to 0.35% in leaves in which CsCCD2L and PaCrtB were co-expressed. Considering that saffron apocarotenoids are costly harvested from flower stigma once a year, and that Buddleja spp. flowers accumulate lower amounts, this system may be an attractive alternative for the sustainable production of these appreciated metabolites.

Topics: Carotenoids; Crocus; Cyclohexenes; Dioxygenases; Glucosides; Nicotiana; Plant Proteins; Plants, Genetically Modified; Potyvirus; Terpenes

The main active components of saffron are crocin, crocetin, picrocrocin, and safranal. There are many studies on their cardioprotective effects, but their cardiotoxicities have not been reported. The human ether-a-go-go-related gene (hERG) K+ channels are of considerable pharmaceutical interest as the target responsible for acquired long QT syndromes. The aim of this study is to explore the effects of crocin, crocetin, picrocrocin, and safranal on the K+ channels encoded by hERG. The interaction of these components with the rapid delayed rectification of K+ currents (IKr) were studied using the perforated patch recording technique. Crocin and picrocrocin had no significant effects on IKr, but crocetin and safranal inhibited hERG K+ currents in a concentration-dependent manner, with IC50 values of 36.35 μM and 37.86 μM, respectively. The maximum inhibitory effects were 37.74 ± 4.14% and 33.74 ± 4.81%, respectively, and the effects were reversible upon washout. The results demonstrate that crocetin and safranal significantly inhibit hERG K+ current, but crocin and picrocrocin do not. This suggests that crocetin and safranal may increase the risk of cardiac arrhythmias by inhibiting IKr.

Topics: Arrhythmias, Cardiac; Carotenoids; Crocus; Cyclohexenes; ERG1 Potassium Channel; Glucosides; Humans; Phytochemicals; Terpenes; Vitamin A

Saffron is the most expensive spice in the world; so, determining optimum conditions for its processing is crucial. The most important processing stage of saffron is drying of its stigma, which should be optimized and there are no reports on reactance-window (RW) of saffron. In this research, drying of saffron with traditional, RW, and oven driers and at three temperatures of 60, 70 and 80 °C, as well as room temperature (25 °C) were studied. Regarding process duration, RW drier with 200 μm Mylar membrane and oven drier were the best methods with average drying time of 25.28 and 22.28 min, respectively. As far as the concentration of bioactive ingredients, i.e., picrocrocin, safranal, and crocin, of saffron was concerned, RW drier with Pyrex glass was better than other driers, resulting in 112.83 [Formula: see text] of picrocrocin, 51.79 [Formula: see text] of safranal, and 274.76 [Formula: see text] of crocin. The panelist most favored those saffron samples dried by RW with 300 μm Mylar membrane.

Topics: Carotenoids; Chemical Phenomena; Color; Crocus; Cyclohexenes; Desiccation; Flowers; Glucosides; Phytochemicals; Powders; Sensation; Spices; Temperature; Terpenes

Crocus sativus L. is an autumn-flowering geophyte belonging to the Iridaceae family, known for the medicinal and coloring uses of the spice from its dried stigmas. It is cultivated in countries with different pedoclimatic conditions. This paper reports on a two-year research project carried out in the Basilicata region of southern Italy on the question of how to obtain the highest performance from saffron. It considers corms from three different geographical origins - 'Sardinia', 'Abruzzo' and 'Kozani' at three different cultivation sites, namely Castelgrande (40° 46' N, 15° 26' E, 781 m a.s.l.), Genzano di Lucania (40° 50' N, 16° 08' E, 344 m a.s.l.), and Villa d'Agri (40° 22' N, 15° 48' E, 638 m a.s.l.).. The highest yields were obtained in the second year by the combination of 'Genzano di L. × Sardinia' and 'Castelgrande × Abruzzo' with 28.1 and 23.9 kg ha. This study demonstrated, through principal component analysis (PCA), that the cultivation site with higher air temperature and without excessive rain during the flowering period generated the best stigma yield with high-quality traits. © 2019 Society of Chemical Industry.

Topics: Carotenoids; Climate; Crocus; Cyclohexenes; Flowers; Glucosides; Italy; Plant Extracts; Quality Control; Rain; Temperature; Terpenes

Saffron is a kind of expensive spice and therapeutic agent. In order to establish a set of methods to evaluate the quality of saffron, High Performance Liquid Chromatography/Mass Spectrometry (HPLC/MS) were used to identify the apocarotenoids extracted from saffron stigmas. TOPSIS method was applied to evaluate the qualities of saffron. The conditions of HPLC/MS and chemical fingerprints of saffron had been established. The 14 chromatographic peaks and 13 main apocarotenoids of saffron with high reproducibility, stability and specificity were confirmed. The qualities of 40 kinds of saffron from different countries and regions were ranked.

Topics: Carotenoids; Chemical Fractionation; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Flowers; Glucosides; Mass Spectrometry; Quality Control; Reproducibility of Results; Solvents; Spices; Terpenes

The lack of an effective vaccine against viral infections, toxicity of the synthetic anti-viral drugs and the generation of resistant viral strains led to discover novel inhibitors. Recently, saffron and its compounds were used to treat different pathological conditions.. In this study, we tested the anti-HSV-1 and anti-HIV-1 activities of Iranian saffron extract and its major ingredients including crocin and picrocrocin as well as cytotoxicity in vitro. The data showed that the aqueous saffron extract was not active against HIV-1 and HSV-1 virions at certain doses (i.e., a mild activity), but crocin and picrocrocin indicated significant anti-HSV-1 and also anti-HIV-1 activities. Crocin inhibited the HSV replication at before and after entry of virions into Vero cells. Indeed, crocin carotenoid suppressed HSV penetration in the target cells as well as disturbed virus replication after entry into the cells. Picrocrocin was also effective for inhibiting virus entry and also its replication.. This monoterpen aldehyde showed higher anti-HSV effects after virus penetrating in the cells. Generally, these sugar-containing compounds extracted from saffron showed to be effective antiherpetic drug candidates.. The recent study is the first report suggesting antiviral activities for saffron extract and its major ingredients. Crocin and picrocrocin could be a promising anti-HSV and anti-HIV agent for herbal therapy against viral infections.

Topics: Animals; Antiviral Agents; Carotenoids; Cell Survival; Chlorocebus aethiops; Crocus; Cyclohexenes; Glucosides; HEK293 Cells; HeLa Cells; Herpesvirus 1, Human; HIV-1; Humans; Plant Extracts; Terpenes; Vero Cells; Viral Plaque Assay; Virus Replication

In this study cold low pressure radiofrequency oxygen plasma was used for the first time to inactivate toxicogenic fungi proliferation on saffron. Varieties of plasma produced reactive oxygen species which were investigated by optical emission spectroscopy. The data were indicative of the absence of UV radiation. Effects of plasma treatment on antioxidant activity, metabolic content, colour, odour and flavour parameters and physical impact on saffron were investigated. A range of plasma powers and exposure times were assayed in suppression of fungal growth. Amongst which power of 60 W for 15 min was used to eradicate Aspergillus and other microorganisms. The ferric reducing antioxidant power was changed from 1778.21 to 1674.25 mM/g dry weight following plasma treatment. Moreover, crocin ester, picrocrocin and safranal metabolites reduced insignificantly. Additionally, plasma had no significant impact on colour, odour and flavour of saffron.

Topics: Antioxidants; Aspergillus; Carotenoids; Crocus; Cyclohexenes; Glucosides; Microscopy, Electron, Scanning; Penicillium; Plant Extracts; Plasma Gases; Rhizopus; Terpenes

The aim of this study was to contribute to the characterization of Crocus taxa using morphological, phytochemical and genetic analysis. The styles of C. cartwrightianus, C. oreocreticus and C. laevigatus, collected in the island of Crete were compared to those of C. sativus cultivated at the region of Western Macedonia (Greece). Phytochemical analysis was done using GC-MS and HPLC methods, while ISSR markers were used for their genetic characterization. Safranal was the major volatile component of the styles of C. sativus, 4-hydroxy-2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde of C. cartwrightianus and C. oreocreticus, and isophorone of C. laevigatus. C. sativus had the highest content of crocins and picrocrocin, while C. laevigatus the lowest (only 5% of C. sativus' quantity) and negligible amount of picrocrocin. According to both the genetic and the chemical analysis, C. cartwrightianus is more related to C. oreocreticus, while C. sativus and C. laevigatus are more distinct. Concordance between the two different types of data was also confirmed by the Mantel test (r = 0.932, P = .68). This is the first thorough screening of secondary metabolites (volatile and non-volatile) and also genetic and morphological characters of wild Crocus styles simultaneously, that contributes to the identification and valorisation of genotypes with similar to C. sativus traits which may be introduced as new cultivars through breeding.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Flowers; Gas Chromatography-Mass Spectrometry; Genetic Markers; Glucosides; Greece; Phytochemicals; Terpenes; Volatile Organic Compounds

This study demonstrates a simple method for one-step isolation of the main secondary metabolites of a hydroalcoholic extract of Crocus sativus stigmas (saffron) using step-gradient centrifugal partition chromatography. The analysis was performed in dual and elution-extrusion mode, using five biphasic systems of the solvents heptane/ethyl acetate/butanol/ethanol/water in ratios of 4:10:0:4:10, 1:13:0:4:10, 1:12:1:4:10, 1:10:3:4:10, and 1:7:6:4:10. Five major crocins, picrocrocin, and crocetin were directly isolated in one step. Scaling up to preparative level, allowed the recovery of significantly high quantities of pure compounds, especially trans-crocin-4, saffron's principal crocin. Comparing dual-mode and elution-extrusion, in dual-mode, the trans-crocin-4 containing fractions were co-eluted with a high amount of free β-d-glucose. In contrast, absence of free β-d-glucose was observed in the corresponding trans-crocin-4 fractions obtained by the second method denoting its superiority against dual-mode. Initiating analysis with the 4

Topics: Acetates; Butanols; Carotenoids; Centrifugation; Chromatography, Liquid; Crocus; Cyclohexenes; Ethanol; Glucosides; Heptanes; Mass Spectrometry; Molecular Structure; Solvents; Terpenes; Vitamin A; Water

The production, characterization and stability of nanoencapsulates of saffron hydrophilic apocarotenoids, i.e. crocins and picrocrocin, in maltodextrin using spray-drying are presented. The effect of mesh size and core:wall ratio (w/w) on the product yield and encapsulation efficiency of these apocarotenoids was examined. Nanoencapsulates were characterized and their stability was examined in the presence or absence of a strong phenolic antioxidant, the caffeic acid, under thermal and in vitro gastrointestinal conditions. Spherical particles were obtained. Product yield and encapsulation efficiency (%) of crocins and picrocrocin was found to be satisfactory. Thermal stability and bioaccessibility of these apocarotenoids was enhanced by nanoencapsulation. Further protection was provided by caffeid acid.

Topics: Antioxidants; Caffeic Acids; Carotenoids; Crocus; Cyclohexenes; Desiccation; Drug Compounding; Drug Stability; Glucosides; Hydrogen-Ion Concentration; Nanocapsules; Plant Extracts; Polysaccharides; Temperature; Terpenes

The present study illustrates a simple and practical way to produce an adequate delivery system of bioactive compounds of saffron by protein-polysaccharide complex. Frist, crocin, safranal, and picrocrocin were loaded in nanodroplets (<100nm) by using water in oil (W/O) microemulsions contain 5, and 10% aqueous saffron extract as a dispersed phase. These microemulsions were then covered with whey protein concentrate (WPC)-maltodextrin or WPC-pectin-maltodextrin through water in oil in water (W/O/W) multiple emulsions. The stability and release of loaded crocin, safranal, and picrocrocin in multiple emulsions were investigated during 22days storage. The produced multiple emulsion by WPC-pectin-maltodextrin along with 5% inner aqueous phase showed a high stability and low release of encapsulated compounds over time. This emulsion also provided a high protection of crocin, safranal, and picrocrocin in the gastric condition.

Topics: Carotenoids; Crocus; Cyclohexenes; Emulsions; Glucosides; Pectins; Plant Extracts; Polysaccharides; Terpenes; Water; Whey Proteins

A microwave-assisted extraction method was optimised for the recovery of bioactive compounds from Crocus sativus L. stigmas with the use of water/ethanol mixture. HPLC-DAD was employed to evaluate the extraction parameters, in particular, solvent type and volume, and the duration of the procedure. Microwave-assisted extraction enhanced the recovery of the active principles, limiting extraction time and solvent waste. Moreover, NIR experiments were performed in order to compare spectra in pseudo-absorbance of Saffron samples with different geographical origins through the application of the chemometric techniques. Moreover, the biological evaluation of crocin 1, safranal and its semisynthetic derivatives as selective inhibitors of five isoforms of human carbonic anhydrase was also explored.

Topics: Carbonic Anhydrase Inhibitors; Carbonic Anhydrases; Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Geography; Glucosides; Humans; Limit of Detection; Plant Extracts; Reproducibility of Results; Spectroscopy, Near-Infrared; Terpenes

Crocus sativus belongs to Iridaceae family and is the only plant species which produces apocarotenoids like crocin, picrocrocin, and safranal in significant quantities. Besides their organoleptic properties, Crocus apocarotenoids have been found to possess remarkable pharmacological potential. Although apocarotenoid biosynthetic pathway has been worked out to a great degree, but the mechanism that regulates the tissue and developmental stage-specific production of Crocus apocarotenoids is not known. To identify the genes regulating apocarotenoid biosynthesis in Crocus, transcriptome wide identification of zinc-finger transcription factors was undertaken. 81 zinc-finger transcription factors were identified which grouped into eight subfamilies. C2H2, C3H, and AN20/AN1 were the major subfamilies with 29, 20, and 14 members, respectively. Expression profiling revealed CsSAP09 as a potential candidate for regulation of apocarotenoid biosynthesis. CsSAP09 was found to be highly expressed in stigma at anthesis stage corroborating with the accumulation pattern of apocarotenoids. CsSAP09 was nuclear localized and activated reporter gene transcription in yeast. It was highly induced in response to oxidative, salt and dehydration stresses, ABA and methyl jasmonate. Furthermore, upstream region of CsSAP09 was found to contain stress and light responsive elements. To our knowledge, this is the first report on the study of a gene family in C. sativus and may provide basic insights into the putative role of zinc finger genes. It may also serve as a valuable resource for functional characterization of these genes aimed towards unraveling their role in regulation of apocarotenoid biosynthesis.

Topics: Arabidopsis; Base Sequence; Carotenoids; Cloning, Molecular; Crocus; Cyclohexenes; Gene Expression Profiling; Gene Expression Regulation, Plant; Genome, Plant; Glucosides; Plant Proteins; Promoter Regions, Genetic; Sequence Analysis, DNA; Terpenes; Transcription Factors; Transcriptional Activation; Transcriptome; Zinc Fingers

One hundred and forty-four Italian saffron samples produced in the years from 2009 to 2015 in five distinct areas located in four different regions, Abruzzo (L'Aquila), Tuscany (Florence), Umbria (Cascia and Città della Pieve) and Sardinia, have been analysed by high-performance liquid chromatography with diode array detection. Intensities of the chromatographic peaks attributed to crocins, safranal, picrocrocin and its derivatives and flavonoids were considered as variables in linear discriminant analysis to attempt geographical classification. The results revealed that spices produced at different sites of the Italian territory can be discriminated with good accuracy. The differentiation of saffron cultivated in Sardinia from those produced in Central Italy was mainly attributed to different contents of the most abundant crocins. Good differentiation of spices produced in close sites of Central Italy was also observed, 88% of validation samples being correctly classified; some minor crocins are responsible for such discrimination.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Discriminant Analysis; Flavonoids; Glucosides; Italy; Plant Extracts; Terpenes

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Topics: Anti-Anxiety Agents; Antioxidants; Biological Products; Cardiovascular Agents; Carotenoids; Color; Crocus; Cyclohexenes; Flowers; Food Contamination; Glucosides; Humans; Odorants; Proteomics; Quality Control; Spices; Taste; Terpenes

Saffron and its components have been suggested as promising candidates for cancer prevention. Carotenoids and monoterpene aldehydes are two potent ingredients of saffron. The goal of the current study was to investigate the anti-tumor effect of chemo-immunotherapy using saffron and its ingredients followed by E7-NT (gp96) DNA vaccine against tumors expressing the E7 protein of human papillomavirus. The in vitro cytotoxic and apoptotic effects of aqueous saffron extract and its components were evaluated in malignant TC-1 and non-malignant COS-7 cell lines. Then, multimodality treatments using E7-NT (gp96) DNA vaccine combined with saffron extract and its ingredients as well as single-modality treatments were tested for their efficacy in inhibiting large and bulky tumor growth. Saffron and its components exerted a considerable anti-tumor effect through prevention of cell growth and stimulation of programmed cell death. Furthermore, 100 % of mice treated with crocin were tumor-free, in contrast to DNA vaccine alone (~66.7 %) and DNA + crocin (~33.3 %) indicating the high potency of crocin as a chemotherapeutic agent. Interestingly, the multimodality treatment using DNA vaccine along with picrocrocin augmented the anti-tumor effects of picrocrocin. Thus, the combination of DNA vaccine with saffron extract and crocin at certain concentrations did not potentiate protective and therapeutic effects compared to mono-therapies for the control of TC-1 tumors.

Topics: Animals; Antineoplastic Agents; Apoptosis; Cancer Vaccines; Carotenoids; Cell Line, Tumor; Chlorocebus aethiops; Combined Modality Therapy; COS Cells; Crocus; Cyclohexenes; Female; Glucosides; Immunotherapy; Mice; Mice, Inbred C57BL; Neoplasms; Papillomavirus E7 Proteins; Papillomavirus Vaccines; Phytotherapy; Plant Extracts; Polyethyleneimine; Terpenes; Transformation, Genetic; Vaccination; Vaccines, DNA

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.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Flowers; Glucosides; Plant Extracts; Quality Control; Species Specificity; Terpenes

Saffron's quality depends on the concentration of secondary metabolites, such as crocins, picrocrocin and safranal. The aim of this research was to evaluate the influence of drying conditions on the secondary metabolite contents of saffron produced in the area of Cascia, in central Italy. Different aliquots of the same saffron sample were subjected to various dehydration conditions and analysed by UV-Vis spectrophotometry to determine crocins, picrocrocin and safranal.. Safranal was also analysed by high resolution gas chromatography, while the crocins and picrocrocin were determined by high-performance liquid chromatography with diode array and mass spectrometric detectors. The results of chromatographic analyses showed that the samples dried in the milder conditions had the lowest content of secondary metabolites. Moreover the sample dried at 60°C for 55min presented the highest contents of trans-crocin-4 and picrocrocin, while safranal was most represented in saffron dried at 55°C for 95min.

Topics: Carotenoids; Chromatography, Gas; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Glucosides; Italy; Plant Extracts; Secondary Metabolism; Terpenes

Saffron (Crocus sativus L.) is one of the most expensive medicinal and aromatic plants in the world. Due to the excessive application of chemical fertilisers in saffron farms and alkalinity of most cultivated soils, split foliar fertilisation has been suggested. The primary goal of this study was to propose split foliar fertilisation as a technique in increasing the quality and quantity of active constituents in saffron. HPLC analysis was used to quantify the most important saffron components; crocins (colour), picrocrocin (taste), and safranal (odour). This study was carried out in Kashmar, Iran, to determine the effect of split foliar fertilisations on quality and quantity of saffron in 2011 and 2012. A split-plot design experiment based on randomised complete block with three replications was conducted to examine three fertiliser types in three regimes for split foliar fertilisation.. Statistical analysis showed that split foliar fertilisation and the fertiliser type significantly increased saffron yield, number of flowers and crocin; whereas it decreased the picrocrocin and safranal content of the saffron stigmas (P < 0.05).. Overall, split foliar fertilisation increased saffron yield and colour but decreased the taste and the odour of saffron.

Topics: Agriculture; Carotenoids; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Fertilizers; Flowers; Glucosides; Humans; Iran; Plant Extracts; Plant Leaves; Terpenes

Saffron, the dried stigmas of Crocus sativus L., is used as a condiment spice. The major bioactive compounds are crocins, picrocrocin and safranal, which are responsible for the sensory profile of saffron (color, flavor and aroma, respectively), and also health-promoting properties. In this paper, the effect on the bioactive compounds of different cooking times in boiling water at 100 °C in samples of Saffron from La Mancha (safranal, picrocrocin, trans-crocin 4, cis-crocin 4 and trans-crocin 3) was investigated. Performance characteristics of High Performance Liquid Chromatography with Variable Wavelength Detector method, parameters of linearity, limits of detection and quantification are reported. High Performance Liquid Chromatography-Photo Diode Array-Mass Spectrometry was used as a confirmatory technique in crocins identification. When the samples are subjected to different cooking times, they present different behaviors, depending on the bioactive compound. In this way, no changes were observed in the concentration of picrocrocin, while heat culinary treatment adversely affects the concentrations of crocins and safranal.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Cooking; Crocus; Cyclohexenes; Flowers; Glucosides; Humans; Mass Spectrometry; Plant Extracts; Spices; Terpenes

The expensive spice saffron originating from the stigmas of Crocus sativus L. and also applied in traditional Chinese medicine (TCM) constitutes a complex mixture of glycoconjugates varying not only in the aglycon structure, but also in glycosylation pattern. Therefore, various tandem mass spectrometric techniques were evaluated for their usefulness in structural elucidation.. Three selected constituents of the stigmas of Crocus sativus L., trans- and cis-crocin-4 as well as picrocrocin, were isolated and purified by HPLC and finally analyzed by ESI-MS (ion trap, QqRTOF), IP-MALDI-MS (QqRTOF) and vMALDI-MS (TOF/RTOF) in combination with tandem mass spectrometry in collision energy regimes ranging from a few eV (LE) to 20 keV (HE) collisions for the first time. These data aid in structurally elucidating minor, unknown glycoconjugates originating from this plant-derived spice.. LE-CID of isomeric crocins on either an ion trap with ESI or a QqRTOF-instrument with ESI or IP-MALDI as desorption/ionization technique only yielded a limited number of structurally diagnostic sodiated product ions related to the carbohydrate moiety as well as to the intact aglycon in contrast to true HE-CID. The low MW constituent picrocrocin did not yield useful LE-CID spectra, but showed a high number of structurally diagnostic product ions by HE-CID utilizing a vMALDI TOF/RTOF-instrument.. The highest number of structurally diagnostic product ions allowing also determination of the carbohydrate linkage of the gentiobiose-moiety of isomeric crocins ((0,4)A(2), (3,5)A(2) product ions indicating a 1→6 carbohydrate linkage) was only achievable by HE-CID. Fragmentation of the aglycon was not observed by any collision energy regime applied.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Crocus; Cyclohexenes; Glucosides; Plant Extracts; Spectrometry, Mass, Electrospray Ionization; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Terpenes

Saffron is an expensive spice, cultivated in many regions of the world. Its chief metabolites include crocins, which are responsible for the coloring ability, safranal, which is the main essential oil constituent, and picrocrocin which is the main bitter constituent of the spice. A simple micellar capillary electrochromatographic (MEKC) method capable of quantifying all three types of main constituents was established. The pH, sodium dodecyl sulphate (SDS) content and electrolyte concentration of the background electrolyte was optimized. A simple extraction protocol was developed which can extract all metabolites of different polarity from the saffron stigmas. Optimal background electrolyte composed of 20 mM disodium phosphate, 5mM sodium tetraborate, 100 mM SDS, pH was set 9.5. Optimal extracting solvent was the background electrolyte, incubated with the sample for 60 min. The proposed method allows quantification of picrocrocin, safranal, crocetin- Di-(β-D-gentiobiosyl) ester and crocetin (β-D-glycosyl)-(β-D-gentiobiosyl) ester within 17.5 min, with limit of detection values ranging from 0.006 to 0.04 mg/ml, from a single stigma.

Topics: Carotenoids; Chromatography, Micellar Electrokinetic Capillary; Crocus; Cyclohexenes; Electrolytes; Glucosides; Hydrogen-Ion Concentration; Limit of Detection; Micelles; Plant Extracts; Sodium Dodecyl Sulfate; Solvents; Terpenes

An ultrasound assisted extraction method is proposed for the recovery of bioactive glycosides (i.e. crocins and picrocrocin) from Crocus sativus L. dry stigmas using aqueous methanol. Response surface methodology (RSM) was employed to optimize the extraction parameters, namely, the percentage of methanol (%), the duration (min) and the duty cycles (s) of sonication. Optical microscopy, spectrophotometry and RP-HPLC-DAD were employed to follow pros and cons of the process. Additional experiments were conducted to compare recoveries with those under other agitation conditions (e.g. magnetic stirring according to ISO 3632-2 standard). The percentage of methanol, the sonication duration and duty cycles combination that can be recommended as optimum for the recovery of crocins and picrocrocin were 50%, 30 min, 0.2s and 0.44%, 30 min, 0.6s, respectively. Picrocrocin levels were not influenced dramatically under the optimum conditions for crocins extraction (11±2 instead of 12±1 mg kg(-1) dry stigmas, respectively) so that these can be considered optimum for both categories of tested compounds. Ultrasound assisted extraction speeded up further recovery of these precious apocarotenoids. Our findings for extraction conditions are useful for both industrial and analytical applications and should be considered in a forthcoming revision of the ISO 3632-2 technical standard.

Topics: Carotenoids; Chemical Fractionation; Chromatography, High Pressure Liquid; Crocus; Cyclohexenes; Flowering Tops; Glucosides; Plant Extracts; Terpenes; Ultrasonics

Extracts from saffron, the dried stigmata from Crocus sativus L., are being used more and more in preclinical and clinical trials for the treatment of cancer and depression. Because of the known quality problems of saffron, HPLC methods on RP(18) 2.5 microm and monolithic RP(18) material have been developed and validated for quality control including the quantification of crocins 1 to 5, crocetin, picrocrocin and the degradation products, the CIS-crocins. Additionally, a GC-MS method has allowed detection and quantification of the volatile compounds from the pentane extract of saffron. Both systems together allowed the comprehensive characterisation of saffron herbal material and extracts for clinical/preclinical trials. For effective preparation of the respective reference standards, a fast centrifugal partition chromatography (FCPC) method was developed allowing the quick isolation of crocins 1, 2, 5 and picrocrocin in good yields. Using these chromatographic methods and the reference standards, a representative survey of saffron from the global market indicated a high variability of quality, especially concerning the amounts of volatile compounds in saffron samples. A specification for high-quality saffron of >20% crocins, >6% picrocrocin and not less than 0.3% of volatiles, calculated as sum of safranal, isophorone and ketoisophorone, was developed. Because no detailed pharmacological studies are available to explain the clinical effects of saffron for the treatment of cancer and depression, receptor binding studies were performed. Saffron extracts and crocetin had a clear binding capacity at the PCP binding side of the NMDA receptor and at the sigma(1) receptor, while the crocins and picrocrocin were not effective. These data could give biochemical support for the above-mentioned pharmacological effects of saffron.

Topics: Carotenoids; Crocus; Cyclohexanones; Cyclohexenes; Flowers; Glucosides; Plant Extracts; Quality Control; Receptors, N-Methyl-D-Aspartate; Receptors, sigma; Reference Standards; Sigma-1 Receptor; Terpenes; Vitamin A

Crocus sativus L. (saffron) has been used as a spice for flavoring and coloring food preparations, and in Chinese traditional medicine as an anodyne or tranquilizer. Our previous study demonstrated that crocin, a carotenoid pigment of saffron, can suppress the serum deprivation-induced death of PC12 cells by increasing glutathione (GSH) synthesis and thus inhibiting neutral sphingomyelinase (nSMase) activity and ceramide formation. The carotenoid pigments of saffron consist of crocetin di-(beta-d-glucosyl)-ester [dicrocin], crocetin-(beta-d-gentiobiosyl)-(beta-d-glucosyl)-ester [tricrocin] and crocetin-di-(beta-d-gentiobiosyl)-ester [crocin]. Saffron also contains picrocrocin, the substance causing saffron's bitter taste. In this study, to confirm whether neuroprotective effects of saffron are caused solely by crocin, we examined the antioxidant and GSH-synthetic activities of these crocins in PC12 cells under serum-free and hypoxic conditions. Measurements of cell viability, peroxidized membrane lipids and caspase-3 activity showed that the rank order of the neuroprotective potency at a concentration of 10 muM was crocin>tricrocin>dicrocin and picrocrocin (the latter two crocins had a little or no potency). In addition, we show that among these saffron's constituents, crocin most effectively promotes mRNA expression of gamma-glutamylcysteinyl synthase (gamma-GCS), which contributes to GSH synthesis as the rate-limiting enzyme, and that the carotenoid can significantly reduce infarcted areas caused by occlusion of the middle cerebral artery (MCA) in mice.

Topics: Animals; Antioxidants; Brain Infarction; Carotenoids; Caspase 3; Cell Hypoxia; Cell Survival; Crocus; Cyclohexenes; Disease Models, Animal; Glucosides; Glutamate-Cysteine Ligase; Glutathione; Infarction, Middle Cerebral Artery; Lipid Peroxidation; Male; Membrane Lipids; Mice; Molecular Structure; Neurons; Neuroprotective Agents; PC12 Cells; Rats; Structure-Activity Relationship; Terpenes; Time Factors; Vitamin A

Extracts of saffron (Crocus sativus L.) have been reported to inhibit cell growth of human tumor cells. In order to study the cytotoxic effect of the characteristic compounds of saffron spice, we have isolated crocin, crocetin, picrocrocin and safranal. Doses inducing 50% cell growth inhibition (LD50) on HeLa cells were 2.3 mg/ml for an ethanolic extract of saffron dry stigmas, 3 mM for crocin, 0.8 mM for safranal and 3 mM for picrocrocin. Crocetin did not show cytotoxic effect. Cells treated with crocin exhibited wide cytoplasmic vacuole-like areas, reduced cytoplasm, cell shrinkage and pyknotic nuclei, suggesting apoptosis induction. Considering its water-solubility and high inhibitory growth effect, crocin is the more promising saffron compound to be assayed as a cancer therapeutic agent.

Topics: Antineoplastic Agents, Phytogenic; Carotenoids; Cell Division; Condiments; Cyclohexenes; Drug Screening Assays, Antitumor; Glucosides; HeLa Cells; Humans; Plant Extracts; Plants, Edible; Terpenes

The determination of saffron components in crude plant extracts by high-performance liquid chromatography-UV-visible photodiode-array detection on-line with mass spectrometry is described. The method is shown to be suitable for the determination of picrocrocin, the glycosidic precursor of safranal, safranal and flavonoids; it is the technique of choice for the analysis of crocetin glycosides (crocins) carrying one up to five glucoses and differentiation of their trans and cis isomers.

Topics: Carbohydrate Sequence; Carotenoids; Chromatography, High Pressure Liquid; Cyclohexenes; Flavonoids; Glucosides; Mass Spectrometry; Molecular Sequence Data; Plant Extracts; Spectrophotometry, Ultraviolet; Spices; Terpenes

High-performance liquid chromatography with photodiode-array detection was used to separate picrocrocin (bitter-tasting component, glucoside of safranal), cis/trans-crocins (carotenoids, glucosyl esters of crocetin) and safranal (flavour, monoterpene aldehyde) of saffron. All components of pure red Greek saffron were extracted from dried stigma with 50% methanol. These compounds were detected, separated collected and identified simultaneously using a Merck LiChroCART 125-4 Superspher 100 RP-18 (4 microns) column and as mobile phase a linear gradient from 20% to 100% acetonitrile in water in 20 min with a detection wavelength at 308 nm.

Topics: Carotenoids; Chromatography, High Pressure Liquid; Condiments; Cyclohexenes; Glucosides; Spectrophotometry, Ultraviolet; Spices; Terpenes