Recovery of ß-carotene from pumpkin using switchable natural deep eutectic solvents.

The aim of the present research was to develop green and sustainable extraction procedure for ß-carotene recovery from pumpkin. A series of hydrophobic natural deep eutectic solvents (NADESs) based onfatty acids were prepared to establish high extraction efficiency of ß-carotene and to increase stability of extracted carotenoids from the pumpkin. To intensify extraction process, NADES composed of C8 and C10 fatty acids (3:1) was selected and coupled with ultrasound assisted extraction. Response surface methodology and artificial neural network model (ANN) model was adopted to analyze significance of extraction parameters demonstrating high prediction levels of the ß-carotene yield, experimentally confirming the maximum ß-carotene content of 151.41 µg/mL at the optimal process condition. Extracted carotenoids in the optimal NADES extract have shown high stability during the storing period of 180 days. A switchable-hydrophilicity eutectic solvent system has been introduced as a successful way to recover extracted carotenoids from the NADES solvent. It was capable of precipitating 90% of carotenoids present in the extract. The proposed procedure is simple, easily scalable and has minimal impact on operators and the environment.

Improvement of Physico-chemical Properties of Arbequina Extra Virgin Olive Oil Enriched with ß-Carotene from Fungi.

Nowadays the consumption of essential carotenoids is reduced due to the lower intake of fruits and vegetables, being humans not capable of synthesizing these molecules. ß-carotene is one of the most important carotenoids possessing anti-oxidation, anti-inflammation and anti-cancer properties. The aim of this work consists of preparing virgin olive oils enriched in ß-carotene from fungi at different concentrations (0.041 and 0.082 mg/mL) in order to obtain new functional foods. Values of quality parameters (free acidity, peroxide value, coefficients of specific extinction and p-anisidine) have been obtained, showing that quality of olive oils was improved. Furthermore, the effect of ß-carotene was evaluated as possible oxidative stabilizer during microwave heating and ultra violet-light exposure of the oils. As expected, the enrichment process brought changes in olive oils color, turning them orange-reddish. The use of natural antioxidants, in particular ß-carotene could be an effective way to protect virgin olive oils from degradation and is a good strategy also to enhance the consumption of bioactive compounds improving olive oils shelf-life and nutritional value.

Identification of ß-carotene oxidation products produced by bleaching clay using UPLC-ESI-MS/MS.

The removal of plant pigments such as ß-carotene is an aspect of vegetable oil processing often desired by the food and pharmaceutical industries. Adsorption of ß-carotene to acid-activated clay (AAC) is a well-established method for purification. Despite this, the removal mechanism of ß-carotene is not well understood. UPLC-MS/MS analysis of surface compounds extracted from ß-carotene-AAC (BC-AAC) complexes show that AAC acts as an oxidiser. Oxidation products detected included canthaxanthin and 3',4'-didehydro-ß-caroten-4-one. AAC had surface water exchanged with an 18O labelled water and was then exposed to ß-carotene. Carotenoids labelled with 18O were produced from this reaction, suggesting surface water is necessary for ß-carotene removal.

A fast and simple method for determination of ß-carotene in commercial fruit juice by cloud point extraction-cold column trapping combined with UV-Vis spectrophotometry.

Cloud point extraction with cold column trapping (CPE-CCT) was used for the rapid preconcentration and UV-Vis spectroscopy of beta-carotene in fruit juice samples. A central composite design was employed to optimize parameters such as pH, incubation time, cloud point temperature and surfactant concentration. A detection limit of 0.01 mg/L of beta-carotene (3SB/m), a coefficient of determination of 0.998 and a linear range of 0.04-10 mg/L were obtained. The CPE-CCT method was confirmed in comparison with the corresponding direct HPLC standard method. A simple, portable and cost-effective device was also utilized. Owing to eliminating centrifugation, the conditions of CPE-CCT were more moderate and its sample handling easier compared to conventional CPE.

Nutritional and Biochemical Characterization of Panus lecomtei Mushroom (Agaricomycetes) from India and Its Cultivation.

Panus lecomtei is emerging as an edible mushroom found worldwide and particularly in the Northern Hemisphere. The mushroom contains a substantial amount of useful nutritional and medicinal compounds. In the present study, we have examined a specimen of P. lecomtei submitted to the ICAR-Directorate of Mushroom Research gene bank. The specimen was examined for taxonomical characters using classical and molecular tools. Attempts were made for cultivation of this mushroom under controlled conditions using sawdust-based substrate. The specimen was characterized by its purplish fruiting body having coarse, rigid, dense hairs on the cap, pubescent stipe, and abundant metuloids. Molecular identification through conserved ITS region was done and the sequence was deposited in NCBI GenBank under accession number MN332200. Nutritional profiling and biochemical analysis showed that the mushroom contained high carbohydrate but low fat contents. The mushroom showed the presence of phenolics, ß-carotene, and lycopene. The analysis also showed substantial antioxidant properties in the mushroom. The findings presented herein point out that P. lecomtei can be used as a potential edible mushroom for diversification of mushroom production in India.

Extraction of Carotenoids from Tomato Pomace via Water-Induced Hydrocolloidal Complexation.

Agro-industrial waste is a largely untapped natural resource of bioactive compounds including carotenoids and pectin. However, conventional solvent extraction involves the excessive use of organic solvents, costly equipment, and tedious operation. These limitations of conventional extraction methods could be prospectively overcome by the carotenoid-pectin hydrocolloidal complexation. The complexation of lycopene and pectin was efficiently promoted in an aqueous environment, resulting in the colloidal complexes that can be subsequently recovered by sedimentation or centrifugation. In this study, the potential of carotenoid-pectin complexation on tomato pomace containing carotenoids and pectin was evaluated. Tomato pomace is a rich source of lycopene, ß-carotene as well as pectin, making it suitable as the raw material for the carotenoid extraction. The extraction of carotenoid and pectin from tomato pomace was optimized using response surface methodology. The maximum recovery was 9.43 mg carotenoid fractions/100 g tomato pomace, while the purity of carotenoid-rich fractions was 92%. The antioxidant capacity of carotenoids extracted from the complexation method was found to be higher than that from the solvent extraction method. Moreover, extraction yield and antioxidant capacity of carotenoid obtained from the carotenoid-pectin complexation were comparable to that from solvent extraction. The carotenoid-pectin complexation is a promising green approach to valorize agro by-products for the extraction of valuable carotenoids.

Improvement of carotenoid extraction from a recently isolated, robust microalga, Tetraselmis sp. CTP4 (chlorophyta).

In recent years, there has been increasing consumer interest in carotenoids, particularly of marine sustainable origin with applications in the food, cosmeceutical, nutritional supplement and pharmaceutical industries. For instance, microalgae belonging to the genus Tetraselmis are known for their biotechnologically relevant carotenoid profile. The recently isolated marine microalgal strain Tetraselmis sp. CTP4 is a fast-growing, robust industrial strain, which has successfully been produced in 100-m3 photobioreactors. However, there are no reports on total carotenoid contents from this strain belonging to T. striata/convolutae clade. Although there are several reports on extraction methods targeting chlorophytes, extraction depends on the strength of cell coverings, solvent polarity and the nature of the targeted carotenoids. Therefore, this article evaluates different extraction methods targeting Tetraselmis sp. CTP4, a strain known to contain a mechanically resistant theca. Here, we propose a factorial experimental design to compare extraction of total carotenoids from wet and freeze-dried microalgal biomass using four different solvents (acetone, ethanol, methanol or tetrahydrofuran) in combination with two types of mechanical cell disruption (glass beads or dispersion). The extraction efficiency of the methods was assessed by pigment contents and profiles present in the extracts. Extraction of wet biomass by means of glass bead-assisted cell disruption using tetrahydrofuran yielded the highest amounts of lutein and ß-carotene (622 ± 40 and 618 ± 32 µg g-1 DW, respectively). Although acetone was slightly less efficient than tetrahydrofuran, it is preferable due to its lower costs and toxicity.

Accumulation of red apocarotenoid ß-citraurin in peel of a spontaneous mutant of huyou (Citrus changshanensis) and the effects of storage temperature and ethylene application.

Red-peeled huyou has a distinct red peel color due mainly to the presence of red apocarotenoid ß-citraurin as well as the increase in amount of total carotenoids. The expression level of carotenoid cleavage dioxygenase 4b1 (CCD4b1) accounted for 99.0% of total transcript abundance of CCD4s in red-peeled huyou peel and was nearly 100 times higher than that in ordinary huyou. ß-Citraurin accumulation and peel coloration was mostly favored at 15 °C but strongly inhibited at moderately high temperatures 20 °C and 25 °C. Exogenous ethylene application for 3 d had no obvious effect on ß-citraurin accumulation in red-peeled huyou but holding fruit at moderately higher temperatures (20 °C and 25 °C) for 3 d had a significant adverse effect on ß-citraurin accumulation. The expression of phytoene synthase 1 (PSY1) and CCD4b1 was higher at 10 °C and 15 °C and significantly lower at 20 °C and 25 °C. The mechanisms governing the accumulation of ß-citraurin are discussed.

Supercritical fluid extraction of ß-carotene from ripe bitter melon pericarp.

Study ascertained the recovery of ß-carotene from enzyme-treated (enzyme load of 167 U/g) pericarp of ripe bitter melon using supercritical fluid extraction (SFE) technique. Effect of different pressure (ranged from 150-450 bar), carbon dioxide (CO2) flow rates (ranged from 15 to 55 ml/min), temperatures (from 50 to 90 °C), and extraction periods (from 45-225 minutes) were observed on the extraction efficiency of ß-carotene. Results showed that extraction pressure (X1) among extraction parameters had the most significant (p < 0.05) effect on extraction efficiency of the ß-carotene followed by allowed extraction time (X4), CO2 flow rate (X2) and the temperature of the extraction (X3). The maximum yield of 90.12% of ß-carotene from lyophilized enzymatic pretreated ripe bitter melon pericarp was achieved at the pressure of approx. 390 bar, flow rate of 35 mL/min, temperature at 70 °C and extraction time of 190 min, respectively. Based on the accelerated storage study the 70% retention shelf life of the ß-carotene into extract was estimated up to 2.27 months at 10 °C and up to 3.21 months at 5 °C.

Preparation and characterisation of novel water-soluble ß-carotene-chitooligosaccharides complexes.

ß-carotene and chitooligosaccharides are bioactive compounds that find their application in the food industry as well in biomedical fields. However, the application of ß-carotene is limited due to its very low water solubility, as well as its air, light and temperature sensitivity. The preparation of ß-carotene-chitooligosaccharides complexes by mechanochemical methods was presented. Their physical and chemical properties including solubility, size, zeta potential and radical scavenging activity were investigated. The interaction of the two components was shown by NMR, FT-IR, and Raman spectroscopy. The complexes were analysed by scanning and transmission electron microscopy. Chitooligosaccharides could serve as a carrier for ß-carotene delivery. The complexation did not cause the loss of the radical scavenging activity of ß-carotene and guaranteed its water solubility.

Mitochondrial Function, Mobility and Lifespan Are Improved in Drosophila melanogaster by Extracts of 9-cis-ß-Carotene from Dunaliella salina.

Carotenoids are implicated in alleviating ageing and age-related diseases in humans. While data from different carotenoids are mixed in their outcomes, those for 9-cis-ß-carotene indicate general positive effects, although basic data on its biological impact are limited. Here, we show that supplementation with 9-cis-ß-carotene in ageing Drosophila melanogaster improved mitochondrial function in terms of ATP production and whole-body respiration and extended mean lifespan. It also resulted in improved mobility. These data provide a potential biological rational for the beneficial effects of dietary supplementation with 9-cis-ß-carotene. These effects may be based on the maintenance of a sound mitochondrial function.

Extraction of Bioactive Compounds Using Supercritical Carbon Dioxide.

Microalgae Dunaliella salina contains useful molecules such as ß-carotene and fatty acids (FAs), which are considered high value-added compounds. To extract these molecules, supercritical carbon dioxide was used at different operative conditions. The effects of mechanical pre-treatment (grinding speed at 0⁻600 rpm; pre-treatment time of 2.5⁻7.5 min) and operating parameters for extraction, such as biomass loading (2.45 and 7.53 g), pressure (100⁻550 bars), temperature (50⁻75 °C) and CO2 flow rate (7.24 and 14.48 g/min) by varying the extraction times (30⁻110 min) were evaluated. Results showed that the maximum cumulative recovery (25.48%) of ß-carotene was achieved at 400 bars and 65 °C with a CO2 flow rate of 14.48 g/min, while the highest purity for stage (55.40%) was attained at 550 bars and 65 °C with a CO2 flow rate of 14.48 g/min. The maximum recovery of FAs, equal to 8.47 mg/g, was achieved at 550 bars and 75 °C with a CO2 flow rate of 14.48 g/min. Moreover, the lowest biomass loading (2.45 g) and the first extraction cycle (30 min) allowed the maximum extraction of ß-carotene and FAs.

Comparative assessment on the extraction of carotenoids from microalgal sources: Astaxanthin from H. pluvialis and ß-carotene from D. salina.

Astaxanthin and ß-carotene are important carotenoids used in numerous pharmaceutical and nutraceutical applications, owing to their vigorous antioxidant properties. The microalgal strains Haematococcus pluvialis and Dunaliella salina accumulate the highest quantities of astaxanthin and ß-carotene (up to 7% and 13% dry weight respectively) and are therefore considered as sustainable feedstock for the commercial production of carotenoids. Thus, from an economical perspective, it becomes desirable to optimize recovery of carotenoids from microalgal cells. To this end, here, we have summarized the conventional and modern extraction techniques generally used for the recovery of astaxanthin from Haematococcus pluvialis and ß-carotene from Dunaliella salina. Furthermore, we have also discussed the optimum process conditions employed for numerous extraction protocols including solvent extraction, ultrasonic-assisted extraction (UAE), microwave-assisted extraction (MAE) and supercritical fluid extraction (SFE). Overall, our study highlights the sustainability of integrated co-production of biofuels and carotenoids in a biorefinery framework.

Biological Active Ecuadorian Mango 'Tommy Atkins' Ingredients-An Opportunity to Reduce Agrowaste.

Mango is a commercially important tropical fruit. During its processing, peel and seed kernel are discarded as waste but they could be recovered as an excellent and cost-effective source of health-promoting ingredients. This study aimed to characterize some of them, including carotenoids like the provitamin A ß-carotene and lutein, with an interest beyond its role in eye health. Other health-promoting compounds like tocopherols and polyphenols were also evaluated, as well as the in vitro antioxidant capacity of mango by-products. Regarding isoprenoids, α-tocopherol was mainly found in the peels and carotenoids concentration was higher in the pulps. ß-carotene was the most abundant carotene in pulp and seed kernel, whereas peel was the only source of lutein, with violaxanthin the most abundant xanthophyll in the different mango organs tested. With regard to polyphenols, peels exhibited greater variability in its phenolic composition, being the total content up to 85 and 10 times higher than the pulp and seed kernels, respectively. On the other hand, peels also stood out for being a very rich source of mangiferin. Seed kernels and peels showed higher antioxidant capacity values than the pulps. These results contribute to the valorization of mango by-products as new natural ingredients for the pharma and food industries.

Identification and biological activity of the volatile compounds of Glycyrrhiza triphylla Fisch. & C.A.Mey.

Chemical composition and biological (antimicrobial, antioxidant and cytotoxic) activities of essential oils (EO) obtained from the aerial parts of Glycyrrhiza triphylla Fisch. & C.A.Mey (G. triphylla) were evaluated in the present study. The EO was isolated and analyzed using gas chromatography-mass spectrometry (GC-MS). Fifty-five compounds representing 99.3% of the total oil composition were identified. Major components of the oil were ß-caryophyllene (25.4%), limonene (16.7%), ß-myrcene (16.0%) and α-humulene (4.4%). The oil composition was dominated by the presence of sesquiterpene hydrocarbons comprising 43.6% of the total oil. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the oil were determined against eight bacterial strains and one fungus. The EO showed a good antibacterial activity against both Gram-positive and Gram-negative bacteria. The most susceptible strain was Micrococcus luteus (MIC = 2.7 µg/mL, MBC = 43.6 µg/mL). The antioxidant potential of the EO was examined using DPPH and ß-carotene/linoleic acid (BCB) assays. The oil was considerably active in the DPPH assay (IC50 = 100.40 ± 0.03 µg/mL). Moreover, in vitro cytotoxic activity was assessed against six cancer cell lines using MTT assay. The EO showed no significant cytotoxic activity. In light of the present findings, G. triphylla oil may deserves to be further investigated for its potential therapeutic effects and also as a natural preservative in food industry.

Variation in Phenolic Profile, ß-Carotene and Flavonoid Contents, Biological Activities of Two Tagetes Species from Pakistani Flora.

The objective of present study was to evaluate the variation in phenolic profile, ß-carotene, flavonoid contents, antioxidant and antimicrobial properties of Tagetes erecta and Tagetes patula (T. erecta and T. patula) through different in vitro assays. Antioxidant activity was determined through 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and inhibition of linoleic acid peroxidation assays and antibacterial and antifungal activities studied using the disc diffusion and resazurin microtiter-plate assays against bacterial and fungal strains. Moreover, total phenolics (TP), total carotenoids (TC) and total flavonoids (TF) were also determined. Highest (TP 35.8 mg GAE/g) and TF (16.9 mg CE/g) contents were found in MeOH extract of T. patula. T. erecta extract showed higher TC contents (6.45 mg/g) than T. patula extract (6.32 mg/g). T. erecta exhibited the highest DPPH radical-scavenging activity (IC50 ) (5.73 µg/mL) and inhibition of linoleic acid peroxidation (80.1%). RP-HPLC revealed the presence of caffeic acid, sinapic acid and ferulic acid in Tagetes extracts, m-coumaric acid in T. erecta whereas chlorogenic acid in T. patula extract only. Both extracts possessed promising antimicrobial activity compared to the ciprofloxacin and flumequine (+ve controls) against Bacillus subtilis and Alternaria alternate. Both extract were rich source of polyphenols exhibiting excellent biological activities.

Adaption of an in vitro digestion method to screen carotenoid liberation and in vitro accessibility from differently processed spinach preparations.

Dark green leafy vegetables are primary food sources for lutein and ß-carotene, however these bioactives have low bioavailability. The effects of mechanical and thermal processing as well as fat addition and fat type on lutein and ß-carotene liberation and in vitro accessibility from spinach were investigated. Lutein liberation and in vitro accessibility were three-fold higher from spinach puree compared to whole leaves. Results for ß-carotene liberation were similar, whereas that of ß-carotene accessibility was only about two-fold. Steaming had no or a negative effect on carotenoid liberation. Fat addition increased ß-carotene liberation from raw and steamed puree, but reduced lutein liberation from steamed leaves and raw puree. Fat types affected ß-carotene differently. Butter addition led to a 2.5 fold increased liberation from raw spinach puree, while the effect of olive and peanut oil was significantly lower, but only minor effects were observed for lutein.

Advancement of green process through microwave-assisted extraction of bioactive metabolites from Arthrospira Platensis and bioactivity evaluation.

Bioactivity and functional properties of cyanobacterial extract mostly depends on process of extraction, temperature and solvent used (polar or non-polar). To evaluate these parameters a design of experiment (DOE; using a 2k design) was performed with Arthrospira platensis. Extraction process was optimized through microwave-assisted extraction considering solvent ratio, temperature and time of extraction with polar (PS) and non-polar (NPS). Maximum extract yield obtained was 4.32±0.25% and 5.26±0.11% (w/w) respectively for PS and NPS. Maximum content of bioactive metabolites in PS extracts were thiamine (846.57±14.12µg/g), riboflavin (101.09±1.63µg/g), C-phycocyanin (2.28±0.10µg/g) and A-phycocyanin (4.11±0.03µg/g), while for NPS extracts were α-tocopherol (37.86±0.78µg/g), ß-carotene (123.64±1.45µg/g) and 19.44±0.21mg/g of fatty acids. A. platensis PS extracts showed high antimicrobial activity and PS extracts had antioxidant activity of 0.79±0.12µmolTE/g for FRAP assay, while for NPS extracts 1.03±0.08µmol α-TE/g for FRAP assay.

In vitro liberation of carotenoids from spinach and Asia salads after different domestic kitchen procedures.

Green-leafy vegetables are rich in nutritionally important constituents including carotenoids. Their potential health benefits depend among others on their liberation from the plant matrix. The aim of the present study was to evaluate the effect of particle size and heat treatments on lutein and ß-carotene liberation from spinach and Asia salads by applying an in vitro digestion protocol and UHPLC analysis. Reduction of particle size resulted in a three- to fourfold increase in liberation of lutein and ß-carotene when comparing whole leaf and puree preparations of spinach. However, this positive effect was shown to be nullified by the severe heat impact during stir-frying of minced spinach, showing that domestic treatments need to be chosen carefully to maximise carotenoid liberation. Steaming significantly improved lutein liberation from Asia salads, but had no or a negative effect in spinach samples, possibly due to differences in liberation or degradation between the two plant matrices.

A novel extraction method for ß-carotene and other carotenoids in fruit juices using air-assisted, low-density solvent-based liquid-liquid microextraction and solidified floating organic droplets.

Green extraction using air-assisted, low-density solvent-based liquid-liquid microextraction and solidified floating organic droplets (AA-LDS-LLME-SFOD) prior to spectrophotometry was successfully applied for quantitation of carotenoids in fruit juices. Under optimal conditions, ß-carotene could be quantified with a linear response up to a concentration of 60 µg mL(-1). The procedure was performed in a microcentrifuge tube with 40 µL of 1-dodecanol as the extraction solvent and a 1.0 mL juice sample containing 8% NaCl under seven extraction cycles of air pumping by syringe. This method was validated based on linearity (0.2-30 µg mL(-1), R(2) 0.998), limit of detection (0.04 µg mL(-1)) and limit of quantification (0.13 µg mL(-1)). The precision, expressed as the relative standard deviation (RSD) of the calibration curve slope (n=12), for inter-day and intra-day analysis was 4.85% and 7.92%, respectively. Recovery of ß-carotene was in the range of 93.6-101.5%. The newly proposed method is simple, rapid and environmentally friendly, particularly as a useful screening test for food analysis.