Recent Advances on the Anticancer Properties of Saffron (Crocus sativus L.) and Its Major Constituents

Cancer is the second leading cause of death globally with an estimated 9.6 million deaths in 2018 and a sustained rise in its incidence in both developing and developed countries. According to the WHO, about 1 in 6 deaths is due to cancer. Despite the emergence of many pioneer therapeutic options for patients with cancer, their efficacy is still time-limited and noncurative. Thus, continuous intensive screening for superior and safer drugs is still ongoing and has resulted in the detection of the anticancer properties of several phytochemicals. Among the spices, Crocus sativus L. (saffron) and its main constituents, crocin, crocetin, and safranal, have attracted the interest of the scientific community. Pharmacological experiments have established numerous beneficial properties for this brilliant reddish-orange dye derived from the flowers of a humble crocus family species. Studies in cultured human malignant cell lines and animal models have demonstrated the cancer prevention and antitumor activities of saffron and its main ingredients. This review provides an insight into the advances in research on the anticancer properties of saffron and its components, discussing preclinical data, clinical trials, and patents aiming to improve the pharmacological properties of saffron and its major ingredients.     

LC Fingerprint and Hierarchical Cluster Analysis of Crocus sativus L. from Different Locations in China

Using simple, sensitive and specific column liquid chromatography, a chemical fingerprinting method was developed for investigating and demonstrating the variance of chemical components among different populations of Crocus sativus L., from 15 locations in China. The LC assay was performed on a reversed-phase C₁₈ column with linear gradient elution using methanol and 1% aqueous acetic acid. The LC data showed considerable variation of chemical constituents among C. sativus populations. Four chemotypes were visually developed from the chromatographic profiles. The grouping of 15 C. sativus populations in hierarchical clustering analysis was in good agreement with the visual comparison of their chromatograms, as demonstrated by chemotypes. In addition, the bioactive compound crocin 1 in this herb was quantitatively determined by a validated reversed-phase LC analysis. Relative peak areas of characteristic compounds were also calculated for quantitative expression of the LC fingerprints. These results showed that the established method for fingerprinting analysis was considered to be suitable for the quality control of C. sativus.     

Bioanalytical Method Development and Validation Study of Neuroprotective Extract of Kashmiri Saffron Using Ultra-Fast Liquid Chromatography-Tandem Mass Spectrometry (UFLC-MS/MS): In Vivo Pharmacokinetics of Apocarotenoids and Carotenoids

Kashmir saffron (Crocus sativus L.), also known as Indian saffron, is an important Asian medicinal plant with protective therapeutic applications in brain health. The main bioactive in Kashmir or Indian Saffron (KCS) and its extract (CSE) are apocarotenoids picrocrocin (PIC) and safranal (SAF) with carotenoids, crocetin esters (crocins), and crocetins. The ultra-fast liquid chromatography(UFLC)- photodiode array standardization confirmed the presence of biomarkers PIC, trans-4-GG-crocin (T4C), trans-3-Gg-crocin (T3C), cis-4-GG-crocin (C4C), trans-2-gg-crocin (T2C), trans-crocetin (TCT), and SAF in CSE. This study’s objectives were to develop and validate a sensitive and rapid UFLC-tandem mass spectrometry method for PIC and SAF along T4C and TCT in rat plasma with internal standards (IS). The calibration curves were linear (R² > 0.990), with the lower limit of quantification (LLOQ) as 10 ng/mL. The UFLC-MS/MS assay-based precision (RSD, <15%) and accuracy (RE, −11.03–9.96) on analytical quality control (QC) levels were well within the acceptance criteria with excellent recoveries (91.18–106.86%) in plasma samples. The method was applied to investigate the in vivo pharmacokinetic parameters after oral administration of 40 mg/kg CSE in the rats (n = 6). The active metabolite TCT and T4C, PIC, SAF were quantified for the first time with T3C, C4C, T2C by this validated bioanalytical method, which will be useful for preclinical/clinical trials of CSE as a potential neuroprotective dietary supplement.     

Eucalyptus gunnii and Eucalyptus pulverulenta ‘Baby Blue’ Essential Oils as Potential Natural Herbicides

The phytotoxicity and eco-compatibility of essential oils (EOs) from Eucalyptus gunnii (EG) and E. pulverulenta ‘Baby Blue’ (EP), cultivated in Italy for their cut foliage, were investigated. Leaf micromorphology, EOs phytochemical characterization, and phytotoxicity were analysed. EP revealed a significantly higher oil gland density and a higher EO yield with respect to EG. In both EOs, 1,8-cineole was the major compound (~75%), followed by α-pinene in EG (13.1%) and eugenol in EP (7.5%). EO phytotoxicity was tested on both weeds (Lolium multiflorum, Portulaca oleracea) and crops (Raphanus sativus, Lactuca sativa, Lepidium sativum, Solanum lycopersicum, Pisum sativum, Cucumis sativus). EG EO inhibited germination of P. oleracea, R. sativus, and S. lycopersicum seeds (ranging from 61.5 to 94.6% for the higher dose used), while affecting only radical elongation in S. lycopersicum (ranging from 66.7 to 82.6%). EP EO inhibited germination of P. oleracea and R. sativus (ranging from 41.3 to 74.7%) and affected radical elongation of L. sativum and L. multiflorum (ranging from 57.4 to 76.0%). None of the EOs affected the germination and radical growing of L. sativa, P. sativum, and C. sativus. Moreover, EP EO was more active than EG EO in inhibiting α-amylase, a key enzyme for seed growth regulation. Brine shrimp lethality assay showed that both EOs are safe for aquatic organisms, suggesting their high eco-compatibility. The data collected provide useful information for future applications of these EOs in agriculture as safe and selective bioherbicides.     

Saffron: Chemical Composition and Neuroprotective Activity

Crocus sativus L. belongs to the Iridaceae family and it is commonly known as saffron. The different cultures together with the geoclimatic characteristics of the territory determine a different chemical composition that characterizes the final product. This is why a complete knowledge of this product is fundamental, from which more than 150 chemical compounds have been extracted from, but only about one third of them have been identified. The chemical composition of saffron has been studied in relation to its efficacy in coping with neurodegenerative retinal diseases. Accordingly, experimental results provide evidence of a strict correlation between chemical composition and neuroprotective capacity. We found that saffron’s ability to cope with retinal neurodegeneration is related to: (1) the presence of specific crocins and (2) the contribution of other saffron components. We summarize previous evidence and provide original data showing that results obtained both “in vivo” and “in vitro” lead to the same conclusion.     

Analysis of Saffron (Crocus sativus L. Stigma) Components by LC–MS–MS

A liquid chromatographic tandem mass spectrometric validated method was developed for the detection of chemicals attributing color, flavor, taste and medicinal properties to saffron (Crocus sativus L. stigma). Ultrasonic extractions of saffron stigmas were followed by LC procedure with Pinnacle II Cyano (5 μm 150 × 2.1 mm) column and acetonitrile: water (70:30, v/v) as mobile phase. Deprotonated ions formed by a turbo ion spray in negative MS mode were used to detect the analytes. MS–MS detection was by monitoring precursors (m/z) fragmentations; of 149 → 113 (safranal), 327 → 283 (crocetin), 329 → 167 (picrocrocin), 355 → 327 (dimethyl crocetin), 489 → 327 (crocin E), 535 → 489 (carotenes), 651 → 327 (crocin C), 813 → 652 (crocin B), 975 → 651 (crocin A) and 1,137 → 813 (crocin F). The method was validated for linearity, precision, repeatability and specificity.     

EPR investigation of some traditional oriental irradiated spices

The 9.50 GHz electron paramagnetic resonance (EPR) spectra of unirradiated and ⁶⁰Co γ-ray irradiated cardamom (Elettaria cardamomum L. Maton, Zingiberaceae), ginger ((Zingiber officinale Rosc., Zingiberaceae), and saffron (Crocus sativus L., Iridaceae) have been investigated at room temperature. All unirradiated spices presented a weak resonance line with g-factors around free-electron ones. After γ-ray irradiation at an absorbed dose of up to 11.3 kGy, the presence of EPR spectra whose amplitude increase monotonously with the absorbed dose has been noticed with all spices. A 100 °C isothermal annealing of 11.3 kGy irradiated samples has shown a differential reduction of amplitude of various components that compose initial spectra, but even after 3.6 h of thermal treatment, the remaining amplitude represents no less then 30% of the initial ones. The same peculiarities have been noticed after 83 days storage at room temperature but after 340 days storage at ambient conditions only irradiated ginger displays a weak signal that differs from those of unirradiated sample. All these factors could be taken into account in establishing at which extent the EPR is suitable to evidence any irradiation treatment applied to these spices.     

Saffron yellow: characterization of carotenoids by high performance liquid chromatography with electrospray mass spectrometric detection

Saffron is one of the oldest natural dyestuffs and is obtained from the dried stigmata of Crocus sativus L. Nowadays, saffron is considered as an invaluable spice of golden‐yellow hue, a precious ingredient in the Eastern and Mediterranean cuisines. It is characterized by a bitter taste that is caused by the chemical properties of its constituents. The yellowness of saffron results from the presence of crocins (glycosyl esters of crocetin), its main color compounds, which are examined in the present study in the crude methanol extracts by high performance liquid chromatography (HPLC) coupled with spectrophotometric and electrospray mass spectrometric detection (HPLC–UV‐Vis–ESI MS). This technique allowed the separation and identification of trans‐ and cis‐isomers of crocins. Their mass spectra registered in the negative ion mode comprised the quasi‐molecular and fragment ions, as well as a range of other ions. Doubly charged ions were found for trans‐isomers only, due to the high symmetry of their molecules. Modification of the eluent allowed the identification of several signals corresponding to adduct ions of crocins with the used additives. Copyright © 2009 John Wiley & Sons, Ltd.     

Synthese von optisch aktiven,natürlichen Carotinoiden und strukturell verwandten Naturprodukten. VI. Synthese von Picrocrocin

Synthesis of optically active natural carotenoids and structurally related compounds. VI. Synthesis of Picrocrocin The synthesis of picrocrocin ( 6 ), the bitter essence of saffron (Crocus sativus L. ) is reported utilizing (3R)-3-hydroxy-β-cyclogeranyl acetate ( 3 ) as a readily available building unit.     

Identification and Isotopic Analysis of Safranal from Supercritical Fluid Extraction and Alcoholic Extracts of Saffron

Abstract

Saffron is one of the most expensive spices. Consequently, it is not so difficult to understand that fraudulent saffron exists. Thus, it was interesting to study the most important flavouring component, in terms of aroma, volatile compound of saffron - safranal - by ¹³C isotopic analysis. Five saffron samples from different countries have been analysed. Safranal has been extracted by methanol or by Supercritical Fluid Extraction (SFE). The results indicate that there is a significant difference between the synthetic safranal and the natural one. On the contrary, it is difficult to conclude on the difference between the various geographical origins, as the isotopic variations are small. Moreover, it has been found that Supercritical Fluid Extraction allowed the selective extraction of volatile compounds from saffron under optimised conditions. It is a cleaner and faster method of extraction compared to the extraction using organic solvent. Nevertheless, an isotopic fractionation occurs in relation to the extraction yield of safranal.     

Effect of ultrasonic waves on crocin and safranal content and expression of their controlling genes in suspension culture of saffron (Crocus sativus L.)

The expression of biosynthesis controlling genes of crocin and safranal in saffron (Crocus sativus) can be influenced by ultrasonic waves. Sterilized saffron corms were cultured in a ½-MS medium supplemented by 2-4-D and BAP. Saffron callus cells were treated with ultrasonic waves in a cellular suspension culture under optimal growth conditions. The samples were collected at 24 and 72 hours after treatment in three replications. The secondary metabolites were measured by high-performance liquid chromatography and the gene expression was analysed by the real-time polymerase chain reaction. Results indicate that this elicitor can influence the expressions of genes CsBCH, CsLYC and CsGT-2; the ultrasonic waves acted as an effective mechanical stimulus to the suspension cultures. The analysis of variance of the ultrasonically produced amounts of safranal and crocin indicates that there is a significant difference between once- and twice-treated samples in that the amount of safranal was the highest within the samples taken from the twice-treated suspension culture at 72 h after the ultrasound treatment, and the crocin was maximised after 24 h passed the twice-applied ultrasound treatment.     

A rapid site‐specific PCR based one‐tube authentication method for saffron in crude drugs and processed herbal medicines

Saffron is one of the world's most precious medicinal herbs and often found to be adulterated with other cheaper materials. The chemical compounds in Crocus sativus L. such as crocin, picrocrocin also exist in other plants, which makes the chemotype‐driven analysis not ideal for the quality control of saffron. Herein, we developed a rapid authentication method for saffron in crude drugs by the site‐specific PCR. In order to realize fast high‐throughput analysis, a one‐tube identification approach was further established by using a universal fluorescent dye to detect the PCR products. In addition, this method was also applied to the authentication of saffron in a processed herbal medicine “Er shi wu wei shan hu wan” which consists of twenty‐five kinds of medicinal materials including plants, minerals and animals. Additionally, this method was also proved to be with high specificity and repeatability. The flexibility of choosing different primers also made this method versatile for other medicinal materials.     

Determination of total safranal by in situ acid hydrolysis in supercritical fluid media: Application to the quality control of commercial saffron

A procedure allowing hydrolysis reactions to be conducted in a dynamic supercritical-CO₂ medium was developed for quantifying total safranal (viz. free safranal present in the sample + safranal resulting from picrocrocin hydrolysis), which are the main component of the essential oil and responsible for the characteristic aroma of saffron. The proposed method allows total safranal amounts over the ranges 0.05-1.5 mg mL⁻¹ to be determined. The standard deviation achieved was 2%. This method was applied to the determination of safranal in natural saffron samples. The results obtained were compared with the “safranal value” total index, which is widely used as a quality measure of saffron products. The comparison revealed that the proposed method provides useful information not contained in the safranal value, based on the fact that, some samples with a high “safranal index” contain low concentrations of safranal. The proposed method is very useful for quality control in commercial saffron samples.     

The GABAA-Benzodiazepine Receptor Antagonist Flumazenil Abolishes the Anxiolytic Effects of the Active Constituents of Crocus sativus L. Crocins in Rats

Anxiety is a chronic severe psychiatric disorder. Crocins are among the various bioactive components of the plant Crocus sativus L. (Iridaceae) and their implication in anxiety is well-documented. However, which is the mechanism of action underlying the anti-anxiety effects of crocins remains unknown. In this context, it has been suggested that these beneficial effects might be ascribed to the agonistic properties of these bioactive ingredients of saffron on the GABA type A receptor. The current experimentation was undertaken to clarify this issue in the rat. For this research project, the light/dark and the open field tests were used. A single injection of crocins (50 mg/kg, i.p., 60 min before testing) induces an anti-anxiety-like effect revealed either in the light-dark or open field tests. Acute administration of the GABA_A-benzodiazepine receptor antagonist flumazenil (10 mg/kg, i.p., 30 min before testing) abolished the above mentioned anxiolytic effects of crocins. The current findings suggest a functional interaction between crocins and the GABA_A receptor allosteric modulator flumazenil on anxiety.     

Phytochemical Profile and Antioxidant Activities of Coleus amboinicus Lour. Cultivated in Indonesia and Poland

Coleus amboinicus Lour., Lamiaceae, is a perennial herb that is native to Indonesia and also cultivated in Africa, Asia and Australia. The major phytochemicals responsible for its bioactivity are rosmarinic acid (RA) and its analogues, flavonoids and abietane diterpenoids. The possibility of cultivation in a colder climate would extend the use of this herb and provide new opportunities to herb growers and livestock farmers. Our study to compare feed value and phytochemical composition of C. amboinicus plants cultivated in its original region, Indonesia, and in Poland. The crude protein content was significantly higher in plants cultivated in Indonesia compared to those cultivated in Poland—21% and 13% of dry matter, respectively. The higher ADF contents were detected in C. amboinicus cultivated in Indonesia, 38–41%, in comparison to 34% in plants cultivated in Poland. The phytochemical composition was also significantly influenced by the cultivation location. Polish samples were higher in polyphenols (RA and its analogues), and also had 1.5–2-fold higher antioxidant potential, as measured by DPPH scavenging, phosphomolybdenum reduction and Fenton reaction driven lipid peroxidation. The Indonesian samples contained more diterpenoid compounds, such as dihydroxyroyleanone, and the sum of terpenoids was ca. 10 times higher than in samples from Poland (15.59–23.64 vs. 1.87 µg/g of extracts). In conclusion, C. amboinicus is suitable for cultivation in non-optimal climatic conditions but some nutritional properties and bioactivity are significantly affected.     

Suppression of Pulmonary Tumor Promotion and Induction of Apoptosis by <Emphasis Type="Italic">Crocus sativus</Emphasis> L. Extraction

Crocus sativus L., commonly known as saffron, is the raw material for one of the most expensive spice in the world, and it has been used in folk medicine for centuries. We investigated the potential of the ethanolic extract of saffron to induce cytotoxic and apoptosis effects in carcinomic human alveolar basal epithelial cells (A549), a commonly used cell culture system for in vitro studies on lung cancer. The cells were cultured in Dulbecco’s modified Eagle’s medium with 10% fetal bovine serum treated with different concentrations of the ethanolic extract of saffron for two consecutive days. Cell viability was quantitated by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Apoptotic cells were determined using annexin V–fluorescein isothiocyanate by flow cytometry. Saffron could decrease the cell viability in the malignant cells as a concentration- and time-dependent manner. The IC₅₀ values against the A549 cell lines were determined as 1,200 and 650 μg/ml after 24 and 48 h, respectively. Saffron-induced apoptosis of the A549 cells in a concentration-dependent manner, as determined by flow cytometry histogram of treated cells that induced apoptotic cell death, is involved in the toxicity of saffron. It might be concluded that saffron could cause cell death in the A549 cells, in which apoptosis plays an important role. Saffron could also be considered as a promising chemotherapeutic agent in lung cancer treatment in future.     

The Common Cichory (Cichorium intybus L.) as a Source of Extracts with Health-Promoting Properties—A Review

Natural products are gaining more interest recently, much of which focuses on those derived from medicinal plants. The common chicory (Cichorium intybus L.), of the Astraceae family, is a prime example of this trend. It has been proven to be a feasible source of biologically relevant elements (K, Fe, Ca), vitamins (A, B₁, B₂, C) as well as bioactive compounds (inulin, sesquiterpene lactones, coumarin derivatives, cichoric acid, phenolic acids), which exert potent pro-health effects on the human organism. It displays choleretic and digestion-promoting, as well as appetite-increasing, anti-inflammatory and antibacterial action, all owing to its varied phytochemical composition. Hence, chicory is used most often to treat gastrointestinal disorders. Chicory was among the plants with potential against SARS-CoV-2, too. To this and other ends, roots, herb, flowers and leaves are used. Apart from its phytochemical applications, chicory is also used in gastronomy as a coffee substitute, food or drink additive. The aim of this paper is to present, in the light of the recent literature, the chemical composition and properties of chicory.     

Pitanga (Eugenia uniflora L.) as a source of bioactive compounds for health benefits: A review

The pitangueira (Eugenia uniflora) is a tree native to Brazil but is cultivated in several subtropical countries. A great diversity of nutrients and bioactive compounds have been found in the leaves and fruits of E. uniflora, which supports its use in folk medicine to treat diseases such as stomach and intestinal disorders, fever and general inflammation. Antimicrobial, antiviral, antifungal and antioxidant effects on metabolism have been reported for this plant. This review discusses the phytochemical profile, toxicity and pharmacological action of E. uniflora leaves and fruits and points out that gaps in the literature that need to be investigated further. This review also discusses studies developed with E. uniflora demonstrating its promising therapeutic potential for several diseases with an apparent low toxicity in mammals. The compilation of the main pharmacological and toxicological results, as well as the phytochemical characterization of the varieties and constituents of E. uniflora are general aspects that this review attempts to demonstrate in order to contribute to the new approaches and developments to plant-derived natural product drug discovery. However, further studies are required to establish the nutraceutical effects and uses of E. uniflora as an important and safe supplement for human health.     

Quality evaluation and quantification of cucurbitacin E in different cultivars of Cucumis sativus L. fruit by a validated high-performance thin-layer chromatography method

Cucumis sativus L. of the Cucurbitaceae family, commonly known as cucumber, is commercially cultivated worldwide. The major phytoconstituents present in the Cucurbitaceae family are different curcurbitacins, principally cucurbitacin E. The content of cucurbitacin E differs within the species or cultivars due to factors like genetic variation and geographical location. The present study reports a simple and rapid quantitative analysis of cucurbitacin E in 5 different C. sativus cultivars by a validated high-performance thin-layer chromatography (HPTLC) method. The mobile phase contained petroleum ether, ethyl acetate and formic acid in the ratio of 40:60:0.5 (V/V). Cucurbitacin E was analyzed densitometrically and the absorbance wavelength was 254 nm. The method showed R_F spot = 0.79 ± 0.06, corresponding to cucurbitacin E in various samples. The calibration curve of standard cucurbitacin E showed good linear relationship in the concentration range of 2‒10 µg/spot with a correlation coefficient (r) > 0.99. The HPTLC method was validated in terms of sensitivity, linearity, accuracy, precision, and specificity as per the International Conference on Harmonization (ICH) guidelines. The present study revealed that the content of cucurbitacin E differs among the C. sativus cultivars. This method may be beneficial for addressing the quality-related aspects of C. sativus for food and pharmaceutical preparation.