Search Results (143)

The quality of Crocus sativus L. flowers, beyond their stigmas, is influenced by the presence of bioactive metabolites also in their floral bio-residues. Given the effect of climatic and soil variables on these bioactive compounds, the aim of this research was to develop an agroecological zoning of saffron crop areas within the Protected Designation of Origin (PDO) La Mancha region (Castilla-La Mancha, Spain) by integrating the floral metabolite content with climatic and soil variables. To achieve this, a total of 173 samples were collected during the 2022 and 2023 harvests and analyzed via RP-HPLC-DAD to determine crocins, picrocrocin, kaempferols, and anthocyanins. Two new indices, Cropi (crocins + picrocrocin) and Kaeman (kaempferols + anthocyanins), were defined to classify flowers into four quality categories (A–D). High-quality classifications (A and B) were consistently associated with plots grouped in the meteorological stations of Ontur, El Sanchón, and Bolaños, indicating favorable edaphoclimatic conditions and climatic parameters, such as moderate temperatures and reduced humidity, for metabolite biosynthesis. In contrast, plots included in the meteorological stations of Tarazona and Pedernoso were mostly assigned to lower categories (C and D). Spatial analysis using thematic maps revealed that areas with an intermediate carbonate content, less calcareous soils, and higher organic matter levels were linked to higher flower quality. These findings highlight the influence of soil characteristics and climate, with distinct seasonal contrasts, that positively influence metabolite synthesis and flower quality. Full article

Crocin biosynthesis involves a complex network of enzymes with biosynthetic and modifier enzymes, and the manipulation of these pathways holds promise for improving human health through the broad exploitation of these bioactive metabolites. Crocins play a significant role in human nutrition and health, as they exhibit antioxidant and anti-inflammatory activity. Plants that naturally accumulate high levels of crocins are scarce, and the production of crocins is highly limited by the characteristics of the crops and their yield. The CCD2 enzyme, initially identified in saffron, is responsible for converting zeaxanthin into crocetin, which is further modified to crocins by aldehyde dehydrogenases and glucosyltransferase enzymes. Crops like tomato fruits, which naturally contain high levels of carotenoids, offer valuable genetic resources for expanding synthetic biology tools. In an effort to explore CCD2 enzymes with improved activity, two CCD2 alleles from saffron and Crocosmia were introduced into tomato, together with a UGT gene. Furthermore, in order to increase the zeaxanthin pool in the fruit, an RNA interference construct was introduced to limit the conversion of zeaxanthin to violaxanthin. The expression of saffron CCD2, CsCCDD2L, led to the creation of transgenic tomatoes with significantly high crocins levels, reaching concentrations of 4.7 mg/g dry weight. The Crocosmia allele, CroCCD2, also resulted in high crocins levels, reaching a concentration of 2.1 mg/g dry weight. These findings underscore the importance of enzyme variants in synthetic biology, as they enable the development of crops rich in beneficial apocarotenoids. Full article

Saffron is a high-cost spice due to the specific conditions for optimal growth and because of being harvested by hand. The massive income from commercializing saffron substituted with other plant parts or low-cost spices makes this spice the main target of fraudsters. Background: Different methods have been developed for detecting saffron adulteration. Most of them are time consuming and complex, and in some types of analysis, the whole untargeted dataset is combined with advanced chemometric tools to differentiate authentic from non-authentic saffron. The official method, combining UV–vis spectroscopy and LC to determine the colour strength and the crocin content, is unable to detect saffron adulterants (safflower, marigold, or turmeric) added at a level lower than 20% (w/w). As a result, innovative approaches based on rapid, high-throughput methods for the identification of adulterated saffron samples are urgently demanded to counteract food frauds. Methods: This paper describes, for the first time, the development of a method combining Direct Analysis in Real Time (DART) with the triple quadrupole MS EVOQ based on the detection of specific MS/MS transitions, promoting a rapid, robust and chromatography-free method capable of monitoring safflower and turmeric adulteration in saffron. Results: The method proved to reach low LODs, allowing the determination of tiny amounts of turmeric and safflower powder in saffron as low as 3% and 5%, respectively, speeding up the whole analytical workflow and enabling us to perform 20 analyses in 10 min. Finally, the greenness of the method was also assessed according to the 0.88 score achieved by submitting it to the greenness calculator AGREE. Conclusions: Given its speed, simplicity, and robustness, this method stands out as a strong candidate for routine implementation in testing laboratories as a rapid screening tool to detect saffron adulteration with safflower or turmeric. Full article

The supplementation of freezing medium with crocin results in an amelioration of post-thawing sperm quality, as determined by motility and viability. This study aimed to examine the molecular mechanisms underlying the ameliorative effect of crocin. Bovine spermatozoa were cryopreserved in a freezing medium supplemented with 0, 0.5, or 1 mM of crocin. Sperm lysates were evaluated for their redox status and the expression of proteins implicated in the heat stress response (HSR) and apoptosis. Crocin protected spermatozoa from the accumulation of superoxide anion and ameliorated their post-thawing antioxidant capacity in terms of ROS scavenging activity and glutathione content. Moreover, crocin decreased the levels of inducible nitric oxide synthase (iNOS), while it increased superoxide dimsutase-1 (SOD-1) levels. These effects were associated with an inhibition of apoptosis, as evidenced by a decreased Bax/Bcl-2 protein ratio and decreased levels of caspase-cleaved substrates. Finally, crocin affected the heat shock response of spermatozoa, since it upregulated the levels of heat shock proteins (Hsp) 60, 70, and 90. In conclusion, the addition of crocin to the freezing medium ensured controlled amounts of ROS, enhanced the antioxidant capacity of spermatozoa, and upregulated the anti-apoptotic proteins and Hsps, thus contributing to the maintenance of cellular homeostasis. Full article

This study aimed to investigate the impact of letrozole and crocin on Leydig cells on busulfan-induced azoospermia using a rat model. A sample population consisting of thirty male rats was randomly assigned to five groups: (1) the control group, (2) azoospermia group, (3) azoospermia group treated with letrozole, (4) azoospermia group treated with crocin, and (5) azoospermia group treated with both letrozole and crocin. The treatment period with letrozole and crocin lasted for four weeks following busulfan administration. Subsequently, comprehensive analyses, including histopathological, molecular, and hormonal assessments, were performed, followed by immunohistochemical staining. This study found that the control group exhibited normal Leydig cell morphology, while the azoospermia group showed reduced Leydig cells and tissue disruptions. Letrozole and crocin treatments were associated with increased testicular fibrosis in the AZO and AZO + Cro groups, while their combination notably reduced fibrosis to approximately 15%. Furthermore, the combination treatment enhanced antioxidant enzyme activity and upregulated androgen receptor expression. Although a number of improvements were noted in sperm motility, they were not statistically significant. Further research is required to clarify the therapeutic implications of these findings in azoospermia treatment. Full article

The economic value of the saffron stigma is primarily due to three crucial apocarotenoids: crocin, picrocrocin, and safranal, which contribute to its color, flavor, and aroma. These compounds make saffron highly valuable in various industries. Plant viruses like the cucumber mosaic virus (CMV) and turnip mosaic virus (TuMV) are significant threats to agricultural crops worldwide, causing economic losses. To elucidate the influence of viral stress on the quality of saffron, morphological, physiological, biochemical, and molecular indexes were assessed. Under the stress of both viruses, typical viral symptoms appeared. The lowest contents of leaf pigments, flowering performance, petal anthocyanin, greenness, and photosynthesis properties were observed in plants infected with CMV and TuMV. According to high-performance liquid chromatography (HPLC) analysis, CMV inoculation led to the highest reduction in crocin and safranal content, while inducing the highest increase in picrocrocin compared to the mock treatment. Gene expression analysis involved in the biosynthesis of crucial secondary metabolites showed a high correlation with the content of each metabolite. CMV inoculation resulted in the lowest expression of CsALDH31l and the highest expression of CsUGT709G1 compared with the mock treatment. Our findings demonstrate the association between virus stress and changes in the metabolism of the saffron medicinal plant. Full article

Saffron petals represent floral biomass generally wasted due to rapid deterioration. Previous characterization studies have revealed the presence of bioactive compounds in petals, such as flavonols and anthocyanins. Petal stabilization is a challenge for the efficient isolation of these compounds. This research evaluated three different drying techniques before the solid–liquid extraction of bioactive compounds: oven-drying (40 and 60 °C), lyophilization, and vacuum evaporation (25 and 50 °C). The characterization of the extracts allowed the annotation of 22 metabolites with a quantitative predominance of anthocyanins and derivatives of kaempferol and quercetin. Oven-drying at 60 °C was the most suitable approach for extracting minor compounds, such as crocins and safranal, at concentrations below 1 mg/g dry weight. Vacuum evaporation (50 °C) and lyophilization were the most recommended strategies for efficiently isolating flavonoids. Therefore, drying saffron petals is crucial to ensure the efficient extraction of bioactive compounds. Full article

Contaminated drinking water is a major health hazard in large urban areas as well as remote communities. Several pollutants detected in rivers and lakes are hormone disruptors that are harmful to consumers as well as aquatic life. In this contribution, we present a new material, synthesized using novel green technologies, designed for solar- or LED-driven degradation of pollutants. This material is based on a glass fiber support, loaded with black TiO₂, a modified form of TiO₂ with strong visible light absorption and without any toxic metal or non-metal dopants. This photocatalyst is fully compatible with flow applications. The effectiveness of the catalyst is demonstrated with crocin and 17β-estradiol, the former being a natural carotenoid used as a screening tool and the latter being a common hormonal disruptor. Our work shows that under visible light illumination, our supported black TiO₂ can degrade these water contaminants with greater efficiency than conventional TiO₂. We envision that our findings can contribute to the production of inexpensive, large-scale solar or LED-based water decontamination systems that could be rapidly deployed to sites in need. Operation of such systems would require minimal training and could be monitored remotely. In addition to the catalyst’s non-toxicity and inflow compatibility, the material also has a long shelf life and is easy and inexpensive to produce, making it an attractive candidate for developing water treatment devices. Full article

The saffron composition is being widely studied for authenticity and traceability, but very few works have been carried out to investigate the relationship between the chemical and physico-chemical properties of saffron solutes and their technological functionality in colloidal systems. This study aims at evaluating the surface properties of saffron extracts obtained using solvents of different polarities to achieve extracts with different compositions in terms of the pattern and content of polar and medium polarity crocins. The air–water surface was evaluated alone and in the presence of Tween 20 at different surfactant–extract ratios. Saffron extracts were able to decrease the surface tension of the aqueous phase, indicating the presence of surface-active compounds. In the mixed saffron extract–Tween 20 systems, competitive adsorption at the air–water interface occurred when the surfactant was present at a low concentration, while at concentrations higher than the CMC, Tween 20 hindered the adsorption of the extract surface-active compounds. The results highlight the interesting technological functionality of saffron extracts for applications in colloidal systems. To better exploit their use in the design and development of formulated foods, nutraceutics and pharma products, further studies are needed to unravel the relationship between the composition of saffron extracts and corresponding surface activity. Full article

Crocus sativus L. is a widely cultivated traditional plant for obtaining dried red stigmas known as “saffron,” the most expensive spice in the world. The response of C. sativus to pre-sowing processing of corms with cold plasma (CP, 3 and 5 min), vacuum (3 min), and electromagnetic field (EMF, 5 min) was assessed to verify how such treatments affect plant performance and the quality and yield of herbal raw materials. The results show that applied physical stressors did not affect the viability of corms but caused stressor-dependent changes in the kinetics of sprouting, growth parameters, leaf trichome density, and secondary metabolite content in stigmas. The effect of CP treatment on plant growth and metabolite content was negative, but all stressors significantly (by 42–74%) increased the number of leaf trichomes. CP3 treatment significantly decreased the length and dry weight of flowers by 43% and 60%, respectively, while EMF treatment increased the length of flowers by 27%. However, longer CP treatment (5 min) delayed germination. Vacuum treatment improved the uniformity of germination by 28% but caused smaller changes in the content of stigma compounds compared with CP and EMF. Twenty-six compounds were identified in total in Crocus stigma samples by the HPLC-DAD method, including 23 crocins, rutin, picrocrocin, and safranal. Processing of Crocus corms with EMF showed the greatest efficiency in increasing the production of secondary metabolites in saffron. EMF increased the content of marker compounds in stigmas (crocin 4: from 8.95 to 431.17 mg/g; crocin 3: from 6.27 to 164.86 mg/g; picrocrocin: from 0.4 to 1.0 mg/g), although the observed effects on growth were neutral or slightly positive. The obtained findings indicate that treatment of C. sativus corms with EMF has the potential application for increasing the quality of saffron by enhancing the amounts of biologically active compounds. Full article

Age-related macular degeneration (AMD) is a common disease contributing to vision loss in the elderly. All-trans-retinal (atRAL) is a retinoid in the retina, and its abnormal accumulation exhibits toxicity to the retina and promotes oxidative stress-induced photoreceptor degeneration, which plays a crucial role in AMD progression. Crocin is a natural product extracted from saffron, which displays significant antioxidant and anti-inflammatory effects. The present study elucidates the protective effects of crocin on photoreceptor cell damage by atRAL and its potential mechanisms. The results revealed that crocin significantly attenuated cytotoxicity by repressing oxidative stress, mitochondrial injury, and DNA damage in atRAL-loaded photoreceptor cells. Moreover, crocin visibly inhibited DNA damage-induced apoptosis and gasdermin E (GSDME)-mediated pyroptosis in photoreceptor cells after exposure to atRAL. It was also observed that crocin distinctly prevented an increase in Fe²⁺ levels and lipid peroxidation caused by atRAL via suppressing the Kelch-like ECH-associated protein 1 (KEAP1)/nuclear factor-erythroid 2-related factor 2 (NRF2)/heme oxygenase-1 (HO-1) signaling pathway, thereby ameliorating photoreceptor cell ferroptosis. In short, these findings provide new insights that crocin mitigates atRAL-induced toxicity to photoreceptor cells by inhibiting oxidative stress, apoptosis, pyroptosis, and ferroptosis. Full article

A predictive model utilizing near-infrared spectroscopy was developed to estimate the loss on drying, total contents of crocin I and crocin II, and picrocrocin content of saffron. Initially, the LD values were determined using a moisture-ash analyzer, while HPLC was employed for measuring the total contents of crocin I, crocin II, and picrocrocin. The near-infrared spectra of 928 saffron samples were collected and preprocessed using first derivative, standard normal variable transformation, detrended correction, multivariate scattering correction, Savitzky–Golay smoothing, and mean centering methods. Leveraging the partial least squares method, regression models were constructed, with parameters optimized through a selective combination of the above six preprocessing methods. Subsequently, prediction models for loss on drying, total contents of crocin I and crocin II, and picrocrocin content were established, and the prediction accuracy of the models was verified. The correlation coefficients and root mean square error of loss on drying, total contents of crocin I and crocin II, and picrocrocin content demonstrated high accuracy, with R² values of 0.8627, 0.8851, and 0.8592 and root mean square error values of 0.0260, 0.0682, and 0.0465. This near-infrared prediction model established in the present study offers a precise and efficient means of assessing loss on drying, total contents of crocin I and crocin II, and picrocrocin content in saffron and is useful for the development of a rapid quality evaluation system. Full article

Crocetin is an aglycone of crocin naturally occurring in saffron and has been proved to have antioxidant, anti-inflammatory, and antibacterial activities. In this experiment, the protective effect of crocetin on vital organs in high-altitude hypoxia rats was studied. Crocetin was prepared from gardenia by the alkaline hydrolysis method, and its reducing ability and free radical scavenging ability were tested. The in vitro anti-hypoxia vitality was studied on PC₁₂ cells. The anti-hypoxic survival time of mice was determined in several models. The acute hypoxic injury rat model was established by simulating the hypoxic environment of 8000 m-high altitude for 24 h, and the anti-hypoxia effect of crocetin was evaluated by intraperitoneal injection with the doses of 10, 20, and 40 mg/kg. The water contents of the brain and lung were determined, and the pathological sections in the brain, lung, heart, liver, and kidney were observed by HE staining. The levels of oxidative stress (SOD, CAT, H₂O₂, GSH, GSH-Px, MDA) and inflammatory factors (IL-1β, IL-6, TNF-α, VEGF) in rat brain, lung, heart, liver, and kidney tissues were detected by ELISA. The results indicated that crocetin exhibited strong reducing ability and free radical scavenging ability and could improve the activity of PC₁₂ cells under hypoxia. After intraperitoneal injection with crocetin, the survival time of mice was prolonged, and the pathological damage, oxidative stress, and inflammation in rats’ tissue were ameliorated. The protective activity of crocetin on vital organs in high-altitude hypoxia rats may be related to reducing oxidative stress and inhibiting inflammatory response. Full article

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. Full article