Quantification Of Carotenoids Research Articles

Grain yellowness is an effective predictor of carotenoid content in global sorghum populations

Identification of high carotenoid germplasm is crucial to assist breeders in provitamin-A biofortification of sorghum (Sorghum bicolor [L.] Moench). High-performance liquid chromatography is the gold standard for carotenoid quantification, however, it is not feasible for large scale phenotyping due to its high cost and low throughput. In this study, we tested the feasibility of using grain color as a high-throughput method of carotenoid biofortification breeding. We hypothesized that visual, color-based selection can be an effective strategy to identify high-carotenoid accessions. Yellow grain had significantly higher carotenoid content than red, brown, and white grain. The degree of yellowness could distinguish the presence or absence of carotenoids, but could not distinguish carotenoid concentrations within yellow-only accessions. The degree of luminosity of the grain, however, was able to better predict carotenoid concentrations within yellow-only accessions. Genome-wide association studies identified significant marker-trait associations for qualitative and quantitative grain color traits and carotenoid concentrations near carotenoid pathway genes—ZEP, PDS, CYP97A, NCED, CCD, and LycE—three of which were common between grain color and carotenoid traits. These findings suggest that using grain color as a method for screening germplasm may be an effective high-throughput selection tool for prebreeding and early-stage breeding in carotenoid biofortification.

Microencapsulated carotenoids from orange sweet potato: Degradation kinetics and physicochemical properties

AbstractThe use of natural pigments in the food industry has increased due to their health benefits. Carotenoids are natural pigments that carry the disadvantage of sensitivity to temperature, light and the presence of enzymes and oxygen. Microencapsulation technology is widely used to protect these compounds against degradation, preserving their physicochemical characteristics. The research objective was to obtain the kinetics of carotenoid degradation of a microencapsulated extract of orange‐fleshed sweet potato (Ipomoea batatas L.) and to establish its physicochemical properties. The sweet potato extract was obtained using ultrasound and then microencapsulated by spray drying using maltodextrin as the wall material. The microencapsulate obtained was subjected to degradation kinetics at 30, 40 and 50°C. Carotenoid quantification analysis, colour, rehydration properties, antioxidant capacity, Fourier‐transform infrared (FTIR) and RAMAN spectroscopy and microstructural characterisation were performed on the microencapsulated carotenoids. From the kinetic analysis, the activation energy of the sweet potato extract was 16.31 kJ/mol while that of the microencapsulated sweet potato extract was 10.27 kJ/mol. Thus, it can be concluded that the microencapsulation process improved the stability of carotenoids subjected to thermal conditions. FTIR and RAMAN spectra confirmed the microencapsulation of the carotenoids. The microencapsulated powder showed a hygroscopicity of 0.22 g/100 g, wettability of 49 s and high solubility. The results obtained in the kinetics are a useful tool to predict losses during the process of heating the carotenoids from sweet potato, thus allowing improvement and selection of the products to which this powder can be applied, in either thermal or cold processes.

Variations of Carotenoids, Total Bioactive Contents, and Antioxidant Activity in Leaves of Medicago polymorpha

Medicago polymorpha L. (MP) is a herbaceous plant commonly known as “bur clover.” It is aboriginal to Central and Western Asia and extensively distributed around the world. This study determines the changes in carotenoids and antioxidant potential of different MP of different origins. The sample leaves were analyzed for identification and quantification of carotenoids using a reversed-phase HPLC system. The result showed nine carotenoids and pigments in each sample. The major carotenoid compounds were phytoene, phytofluene, neoxanthin and its isomer (9-Z-neoxanthin), violaxanthin, lutein along with their isomers (9-Z-lutein, 9′-Z-lutein), and all-E-β-carotene. The major pigments were 15-hydroxy-lactone chlorophyll a, pheophytin a, pheophytin a′, pyropheophytin a, divinyl chlorophyll a, chlorophyll b′, chlorophyll b, 13′-hydroxy-lactone chlorophyll b, and hydroxy pheophytin a′. The carotenoids were detected in considerable amounts in the samples from Lower Dir (213 μg/g), Swat (171 μg/g), and Buner (157 μg/g). Chlorophylls were higher in Lower Dir (203.4 μg/g), Swat (184.0 μg/g), and Buner (152.2 μg/g) and significantly lower amounts in Malakand samples (141.7 μg/g). The total carotenoids in Lower Dir (51.2%) were higher than in Swat (48.2%), Buner (50.8%), and Malakand samples (44.6%). The amounts of violaxanthin and lutein were significantly higher in Lower Dir (73.6%) samples, followed by Malakand (51.7%), whereas Buner and Swat samples showed the lowest levels. In conclusion, MP leaves are a good source of important carotenoids having potential antioxidant properties, which are highly correlated to the violaxanthin and lutein contents.

Advancing carotenoid Quantification: A new method for semi-quantitative assessment of β -Carotene and lycopene content in food extracts

Carotenoids, such as lycopene and β-carotene, have been widely recognized for their antioxidant properties and potential health benefits. Accurate quantification of carotenoids in plant extracts is essential for nutritional assessment, quality control, and research investigations. This study introduces an innovative method for quantifying lycopene and β-carotene, in plant extracts and aims to bridge the gap between complex and expensive carotenoid quantification techniques and the need for accessible methods that can be widely adopted. The primary difference between HPLC and HPTLC lies in the medium used for separation. HPLC employs a liquid phase within columns, while HPTLC utilizes a thin layer of adsorbent on a plate. This distinction impacts factors like equipment, cost, and analysis time. The VisionCats software, combined with the CAMAG Visualizer-2, allows the semi-quantification of metabolites using an image-based evaluation method enabling the simultaneous assessment of qualitative and semi-quantitative information from the HPTLC images.Sample preparation involves washing and drying the vegetal material, followed by dichloromethane extraction. HPTLC analysis is performed using the CAMAG Advanced Herbal System, and the validation studies include establishing calibration curves and determining the detection threshold and minimum quantification threshold for lycopene and β-carotene. Specificity and precision were evaluated to ensure accurate identification and repeatability of the method. Data analysis involves selecting the regression method based on the nature of the data and assessing the goodness of fit using the R2 value. The results showed distinct peaks corresponding to lycopene and β-carotene in the chromatograms of the plant extract samples. The visualizer-based method demonstrates good specificity and precision, with no interfering peaks observed and low relative standard deviation. The method shows promising results regarding specificity, precision, and reliability. It has the potential for broader implementation in carotenoid research and rapid monitoring of carotenoid content in various agricultural and food products, particularly in resource-limited settings. Further optimization and validation on a wider range of samples would enhance the applicability of this method in carotenoid research.Sample preparation involves washing and drying the vegetal material, followed by dichloromethane extraction. HPTLC analysis is performed using the CAMAG Advanced Herbal System, and the validation studies include establishing calibration curves and determining the detection threshold and minimum quantification threshold for lycopene and β-carotene. Specificity and precision were evaluated to ensure accurate identification and repeatability of the method. Data analysis involves selecting the regression method based on the nature of the data and assessing the goodness of fit using the R2 value. The results showed distinct peaks corresponding to lycopene and β-carotene in the chromatograms of the plant extract samples. The visualizer-based method demonstrates good specificity and precision, with no interfering peaks observed and low relative standard deviation. The method shows promising results regarding specificity, precision, and reliability. It has the potential for broader implementation in carotenoid research and for rapid screening and monitoring of carotenoid content in various agricultural and food products, particularly in resource-limited settings. Further optimization and validation on a wider range of samples would enhance the applicability of this method in carotenoid research.

Thermal Treatment and High-Intensity Ultrasound Processing to Evaluate the Chemical Profile and Antioxidant Activity of Amazon Fig Juices

The present paper evaluated the influence of heat treatment (HT) and high-intensity ultrasound (HIUS) on the chemical profile of the Amazon fig (Ficus subapiculata, Moraceae) juices. Antioxidant activity, quantification of carotenoids, total phenolic compounds (TPC), pH, titratable acidity, soluble solids, color and chemical profile (NMR) were evaluated. Treatments did not change the pH (3.4–3.5), titratable acidity (0.044–0.048%) and soluble solids (2.3–2.4 °Brix). The highest antioxidant activity (DPPH, ABTS) and TPC were presented by the HT-treated juice, which was equivalent to 1235 ± 11 µM TE, 1440 ± 13 µM TE and 312 ± 5 mg GAE mL−1, respectively. The treatments influenced the color luminosity according to the L* and a* parameters, while the b* parameter showed no significant change. The L* parameter was elevated in all treated samples compared to the control sample. Analyzing the parameter a* f, it was verified that the sample with thermal treatment (HT) was different from the control sample, but presented similarity with the samples of the HIUS processes. The 1H NMR spectra of the juices showed similar chemical profiles in all treatments. The compounds α-glucose, β-glucose, fructose, citric, malic, quinic, and p-hydroxybenzoic acids were identified. The HT treatment presented higher efficiency to extract the antioxidant compounds from fig juices. The HIUS treatments with constant energy density also improved the tolerance of the antioxidant compounds, especially in conditions of higher potency and reduced time. Future studies will be devoted to carry out microbiological analysis and evaluate the stability of treated juices.

Evaluation of Guava Pulp Microencapsulated in Mucilage of Aloe Vera and Opuntia ficus-indica as a Natural Dye for Yogurt: Functional Characterization and Color Stability.

The substitution of artificial colorants for pigments extracted from fruits is a highly desirable strategy in the food industry for the manufacture of natural, functional, and safe products. In this work, a 100% natural spray-dried (SD) microencapsulated colorant of pink guava pulp, using aloe vera (AV) or Opuntia ficus-indica (OFI) mucilage as functional encapsulating material, was prepared and evaluated as an additive into a yogurt (Y) matrix. The characterization of yogurt samples supplemented with OFI (Y-SD-OFI) and AV (Y-SD-AV) mucilage-covered guava pulp microcapsules was carried out through carotenoid quantification using UV–vis and HPLC–MS techniques, dietary fiber content, antioxidant capacity, colorimetry, and textural analysis, as well as by an evaluation of color stability after 25 days of storage at 4 °C in the dark. These physicochemical characteristics and color stability on the Y-SD-OFI and Y-SD-AV samples were compared with those of a commercial yogurt (control sample, Y-C) containing sunset yellow FCF synthetic colorant (E110). Y-SD-OFI and Y-SD-AV samples exhibited a high content of lycopene, dietary fiber, and antioxidant activity, which were absent in the control sample. Microencapsulated lycopene imparted a highly stable color to yogurt, contrary to the effect provided by the E110 dye in the control sample. The texture profile analysis revealed an increase in firmness, consistency, and cohesion in the Y-SD-OFI sample, contrary to the Y-SD-AV and Y-C samples, which was attributed to the variation in fiber concentration in the microcapsules. The incorporation of OFI and AV mucilage microparticles containing pink guava pulp into yogurt demonstrated its potential application as a functional natural colorant for dairy products.

Identification, Quantification and Kinetic Study of Carotenoids and Lipids in Rhodotorula toruloides CBS 14 Cultivated on Wheat Straw Hydrolysate

Production of carotenoids and lipids by Rhodotorula toruloides CBS 14 cultivated on wheat straw hydrolysate was investigated. An ultra-high-performance liquid chromatography (UHPLC) method for carotenoid quantification was developed and validated. Saponification effects on individual carotenoid quantification were identified, and lipid and carotenoid kinetics during cultivation were determined. The carotenoids β-carotene, γ-carotene, torularhodin, and torulene were identified; β-carotene was the major carotenoid, reaching a maximum of 1.48 mg/100 g dry weight. Recoveries of the carotenoids were between 66% and 76%, except torulene and torularhodin, which had lower recoveries due to saponification effects. Total carotenoid content in saponified and unsaponified yeast extract, respectively, determined by UHPLC or photometer, respectively, was 1.99 mg/100 g and 4.02 mg β-EQ/100 g dry weight. Growth kinetics showed a positive correlation between carotenoid content and lipid accumulation. β-carotene was the major carotenoid at all time points. At the end of the cultivation, triacylglycerols (TAGs) were the major lipid class, with 58.1% ± 3.32% of total lipids. There was also a high proportion of free fatty acids, reaching from 20.5% to 41.8% of total lipids. Oleic acid (C18:1) was the major fatty acid. The lipid yield at the end of the cultivation was 0.13 g/g of sugar consumed.

An optimal saponification and extraction method to determine carotenoids in avocado

Quantification of carotenoids in avocado fruit is a great challenge due to their co-extraction with high-oil concentration and the inherent nature of carotenoids to degrade and undergo cis/trans photoisomerization with prolonged extraction times and high temperatures. The study provides an optimised and validated methodology for quantification of carotenoids in the high-oil avocado matrix, with > 93% recovery of all carotenoids tested being significantly greater than previously published. Saponification with 15% KOH for 60 min was optimal for the avocado matrix. For the first time, this study identified that soap, produced during the saponification reaction, resulted in a significant reduction of carotenoid content from the avocado matrix, due to the production of micelles. A significantly higher carotenoid content (3.58 versus 2.0 mg/100 g DW) was able to be extracted after saponification with acidified phosphate buffer, instead of water as reported previously. Carotenoid profiles of five avocado cultivars were identified and quantified.

Impact of cooking on the bioactive compounds and antioxidant activity of gherkin (Cucumis anguria L.)

Despite the high consumption of Japanese gherkin in the Northeast of Brazil, very few studies with this vegetable are found in the literature. As it is consumed raw or after cooking, being mainly consumed cooked, there was the interest to know if cooking could influence bioactive compounds content and antioxidant activity since high temperatures are related to benefits losses in food. This research aimed to compare the content of bioactive compounds and the antioxidant activity between in natura and cooked Japanese gherkin. Gherkins were submitted to quantification of carotenoids, anthocyanins, vitamin C and phenolic compounds, in addition to the antioxidant activity analyses through the ferric reducing antioxidant power (FRAP) and the DPPH free radical scavenging methods. After cooking, carotenoids (from 4.90 to 4.59 ?g.g-1) and phenolic compounds (from 17.07 to 14.95 mg.100 g-1) content decreased, but with no significant difference between in natura and cooked samples. As for the antioxidant activity, the in natura gherkin showed a higher value (3.31 ?M Ferrous Sulfate.g-1) than the cooked one (1.96 ?M Ferrous Sulfate.g-1) for FRAP method, while for DPPH method no difference between the samples was detected. Anthocyanin and vitamin C were not detected in gherkins. We concluded that, in general, cooking did not affect the bioactive compounds of gherkins and that antioxidant activity increased.

Different spectrophotometric methods for simultaneous quantification of lycopene and β-carotene from a binary mixture

UV-VIS spectrophotometric analysis is one of the cheapest and most handy methods used for carotenoids quantification in solutions. According to Lambert-Beer's law, carotenoids concentrations in a mixture are computed using mixture absorbances and characteristic absorption coefficients for pure compounds. This work proposes four methods for simultaneous quantification of two carotenoids using mixture spectral data, physical models of pure and mixtures spectra in acetone-hexane mixture, different wavelengths groups for absorbance of each carotenoid, the spectra overlapping property and isosbestic point presence. Two absorbance factors defined to improve the concentrations equations accuracy and different assumptions in terms of wavelengths groups and contribution of each carotenoid at mixture absorbance are considered. These methods are validated using synthetic mixtures for two carotenoids (lycopene and β-carotene) and two tomato residue extracts using equivalence test as statistical tool. IPM–II–WG6 method gives good correlation from statistical point of view, with medium accuracy errors.

Feasibility of Individual Carotenoid Quantification in Mixtures Using UV-Vis Spectrophotometry with Multivariate Curve Resolution Alternating Least Squares (MCR-ALS)

A fast and low-cost analytical method to determine the concentrations of carotenoids (β-carotene, lutein, and lycopene) from mixed standard solutions or from fruit extracts (kiwi fruits, tomato paste, pink grapefruit juice, kiwi-pineapple smoothie, and apricot nectar) was tested. The methodology was based on UV-Vis spectrophotometry and Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS). Results showed that relative concentrations of β-carotene, lutein, and lycopene in solvent were successfully determined by this technique with an error inferior to 6%. In real extracts, the procedure succeeded well in identifying the major carotenoid type of the fruit samples but also a more complex profile of a fruit mixture. The results also showed that accuracy of carotenoids determination by UV-Vis spectrophotometry-MCR-ALS in fruit extracts was conditioned by their spectral characteristics (III/II ratios and λmax), their relative proportion, and the extract purity.

Engineering the polyphenolic biosynthetic pathway stimulates metabolic and molecular changes during fruit ripening in "Bronze" tomato.

The metabolic engineered Bronze tomato line is characterized by the constitutive over-expression of the VvStSy gene encoding a structural protein responsible for the stilbenoids biosynthesis and the fruit-specific over-expression of AmDel/Rosea1 and AtMYB12 genes encoding transcription factors that activate the polyphenol biosynthetic pathway. This tomato line is known for the increased levels of polyphenols in ripe fruits and for beneficial health promoting antioxidant and anti-inflammatory effects. In this study we analyzed the transcriptional and metabolic profiling in mature green, breaker, orange and ripe fruits compared to the normal tomato counterparts during ripening, to unravel the effect of regulatory and structural transgenes on metabolic fluxes of primary and secondary metabolisms. Our results showed that the shikimate synthase (SK) gene was up-regulated in the Bronze fruit, and the transcriptional activation is consistent with the metabolic changes observed throughout fruit ripening. These results paralleled with a reduced level of simple sugars and malate, highlighting the consumption of primary metabolites to favor secondary metabolites production and accumulation. Finally, carotenoids quantification revealed a change in the lycopene/β-carotene ratio in the Bronze fruit as a consequence of significant lower level of the first and higher levels of the latter. The high polyphenols and β-carotene content displayed by the Bronze fruit at the later stages of fruit ripening renders this line an interesting model to study the additive or synergic effects of these phyto-chemicals in the prevention of human pathologies.

Laser capture of tomato pericarp tissues for microscale carotenoid analysis by supercritical fluid chromatography.

Plant organs and tissues are comprised of an array of cell types often superimposed on a gradient of developmental stages. As a result, the ability to analyze and understand the synthesis, metabolism, and accumulation of plant biomolecules requires improved methods for cell- and tissue-specific analysis. Tomato (Solanum lycopersicum) is the world's most valuable fruit crop and is an important source of health-promoting dietary compounds, including carotenoids. Furthermore, tomato possesses unique genetic activities at the cell and tissue levels, making it an ideal system for tissue- and cell-type analysis of important biochemicals. A sample preparation workflow was developed for cell-type-specific carotenoid analysis in tomato fruit samples. Protocols for hyperspectral imaging of tomato fruit samples, cryoembedding and sectioning of pericarp tissue, laser microdissection of specific cell types, metabolite extraction using cell wall digestion enzymes and pressure cycling, and carotenoid quantification by supercritical fluid chromatography were optimized and integrated into a working protocol. The workflow was applied to quantify carotenoids in the cuticle and noncuticle component of the tomato pericarp during fruit development from the initial ripening to full ripe stages. Carotenoids were extracted and quantified from cell volumes less than 10nL. This workflow for cell-type-specific metabolite extraction and quantification can be adapted for the analysis of diverse metabolites, cell types, and organisms.

Purification and use of carotenoid standards to quantify cis-trans geometrical carotenoid isomers in plant tissues.

Reverse-phase high-performance liquid chromatography (HPLC) is a preferred method used to identify and quantify carotenoids. Here, we describe a straightforward, reliable, and cost-effective protocol to purify and develop individual carotenoid standards for absolute quantification of carotenoids, including selected cis-trans (geometric) isomers. Analytical techniques to extract, purify and collect individual carotenoids using an HPLC system equipped with a Diode Array Detector (DAD) and fraction collector are described. Carotenoids were separated and identified by their characteristic ultraviolet-visible (UV-Vis) absorption spectra and individually isolated based on their retention times using a C30 column. This chapter outlines how to prepare standard calibration curves using known quantities of purified and/or commercially available carotenoids. A series of molar extinction and slope coefficients for phytoene, phytofluene, ζ-carotene, neurosporene, pro-lycopene, all trans-lycopene, lutein, β-carotene, zeaxanthin, antheraxanthin, violaxanthin, neoxanthin, capsanthin, capsorubin and β-cryptoxanthin are defined to enable absolute quantification of their abundance in plant, animal, and bacterial tissues. Different approaches for reporting carotenoid abundance by absolute concentration, relative composition, and/or using ratios of different pigments are provided as a convenient resource for carotenoid researchers.

HPLC analysis of carotenoids from bacteria.

Bacteria synthesize different carotenoid structures. A powerful method for the determination of carotenoid profiles of bacteria, including quantification, is high performance liquid chromatography (HPLC). This chapter explains the basics of this method and its application, and provides instruction on how to proceed with HPLC. A basic HPLC system for bacterial carotenoids is presented which allows tentative identification and quantification. The individual analysis steps including extraction, sample preparation and HPLC separation are described in detail. HPLC examples of structurally diverse carotenoids from different bacterial phyla are displayed demonstrating the applicability of this method. In addition, information on spectroscopic properties useful for identification is included. Finally, a procedure for carotenoid quantification from the HPLC chromatogram is described.

Extraction of Lipophilic Antioxidants from Native Tomato Using Green Technologies.

SUMMARYResearch backgroundTomato (Solanum lycopersicum L.) fruit is highly consumed worldwide and contains high amounts of carotenoids and tocopherols, two powerful antioxidants. Native tomato genotypes are rarely used in large-scale market but serve as a reservoir to diversify the species gene pool and can be employed to obtain functional compounds. Extraction methods are currently changing towards cleaner procedures that are more efficient and environmentally friendly, including avoiding toxic or polluting solvents.Experimental approachIn this study, factorial and fractional factorial designs were used to evaluate the efficiency of digestive enzymes, sonication and green solvents to obtain lipophilic antioxidant extracts from native tomato. To monitor the efficiency of the extraction process, spectrophotometric quantification of total carotenoids and antioxidant activity was carried out, and then individual quantification of carotenoids and tocopherols in the extracts was done by HPLC.Results and conclusionsDigestive enzymes and sonication increased the carotenoid content and the antioxidant activity of the obtained extracts when applied individually. However, when these treatments were applied together and in combination with isopropyl acetate, a green solvent, the obtained extracts had the highest carotenoid and tocopherol contents as well as the maximal antioxidant activity. A correlation analysis suggested that antioxidant activity resulted from synergistic effects rather than individual compounds. Tomato extracts were obtained through a rapid and environmentally friendly extraction method and their antioxidant activity was enhanced.Novelty and scientific contributionTomato fruits have been the subject of numerous studies; however, functional compound extraction through environmentally friendly methods remains an attractive use of native tomato fruit, enhancing its limited production and harnessing a large amount of tomato product industry. There are few reports where environmentally friendly extraction methods are combined; even rarer are those where green solvents are also used. In this work, the combination of different environmentally friendly extraction methods improved the extraction of carotenoids and tocopherols and allowed to establish a more efficient process. These results could stimulate the use of clean technologies and make the native tomato more attractive for industrial or compound extraction processes.

Resonance Raman and SERRS of fucoxanthin: Prospects for carotenoid quantification in live diatom cells

Fucoxanthin, one of the most abundant carotenoids in brown algae or in unicellular diatoms, plays a crucial role in photosynthetic processes and has a significant contribution to the natural pigments industry and biomedical application as colouring and bioactive agent. Many carotenoids are widely identified using resonance Raman (RR) spectroscopy, however, quantitative estimation of this pigment in solutions and in live cells from RR data is challenging. Here we investigated the concentration dependence resonance Raman and surface-enhanced resonance Raman scattering (SERRS) spectra of fucoxanthin, to prospect the capability of the techniques for quantitative estimation of the carotenoid in live diatom species, Cylindrotheca closterium and Skeletonema costatum. Furthermore, since pools of carotenoids are usually present in live photosynthetic microorganisms, we compared their SERRS signal when exposed to Ag nanoparticles and found distinct SERRS carotenoids signature. Regarding fucoxanthin quantification, we found a nonlinear increase in the resonance Raman intensity of the ν(C=C) mode in the concentration range from 1.190 µM to 3.035 mM and a linear increase in SERRS intensity in the 0.12 to 1.186 µM concentration range, comparable with the signal intensity collected from single diatom cells. Particularly, S. costatum, an opportunistic diatom causing harmful bloom in certain conditions, could provide important information from carotenoids monitoring in relation with its adaptability to various environmental factors, which would certainly consolidate such monitoring approaches relying on Raman spectroscopy techniques.

Chemical Profile and Evaluation of the Antioxidant and Anti-Acetylcholinesterase Activities of Annona squamosa L. (Annonaceae) Extracts.

This study presents the chemical profile of extracts from the pulp and seed of Annona squamosa L., as well as the evaluation of their antioxidant and acetylcholinesterase inhibition activities. In the chemical prospection, qualitative assays were performed, and the contents of total phenols, flavonoids, vitamin C, and carotenoids were quantified. For the compounds identification, analyses of the extracts were performed by liquid chromatography coupled to mass spectrometry. Antioxidant evaluation was performed using the DPPH, ABTS, Fe3+ reduction, 2-DR protection, and β-carotene protection methods. The assay for inhibition of acetylcholinesterase activity was determined using the method described by Ellman. The secondary metabolites identified were anthocyanidins, flavones, flavonols, and alkaloids. Phenol analysis showed a higher quantitative value of total phenols and flavonoids for the seed extract, and the vitamin C content was higher in the pulp extract. There was no significant difference in relation to the carotenoids quantification. The best results obtained for antioxidant activity, for both seed and pulp extracts, were with the ABTS method with IC50 of 0.14 ± 0.02 and 0.38 ± 0.02 mg/mL, respectively. Compared to A. squamosa seed extract, the pulp extract demonstrates higher AChE inhibitory activity with IC50 of 18.82 ± 0.17 µg/mL. A. squamosa is a nutritious food source. The continuity of the studies is fundamental to relate the consumption of this food and its effects on neurodegenerative diseases.

Assessing the Fatty Acid, Carotenoid, and Tocopherol Compositions of Seeds from Apple Cultivars (Malus domestica Borkh.) Grown in Norway.

Apple production generates large amounts of apple pomace including seeds, leading to high transportation costs, public health hazards and undesirable odor. A new reuse strategy of this kind of waste could solve environmental issues and/or create unconventional sources of health beneficial products. In total, seeds from 75 apple cultivars grown in Norway (both domestic and international) have been analyzed for the first time for oil content and fatty acid profile together with tocopherols and carotenoids quantification in defatted seeds. Seeds from cultivar Håkonseple had the highest oil content (22.10%), with linoleic, oleic acid, and palmitic acid as the most abundant fatty acids. The levels of β-carotene and lycopene carotenoids and α-tocopherol were the highest in defatted seeds of the cultivar Sureple Grøn. Principal component analysis separated cultivars according to the total oil content. The Norwegian apple cultivars Håkonseple, Kviteple, Tolleivseple, Vinterrosenstrips, and Tokheimseple are recommended for obtaining vegetable oil due to their high oil contents, while cultivar Sureple Grøn can be separated due to its high levels of β-carotene, lycopene and total tocopherols.

Effect of different levels of synthetic astaxanthin on growth, skin color and lipid metabolism of commercial sized red porgy (Pagrus pagrus)

A study was undertaken to evaluate the effects of supplementing commercial feeds with Carophyll® Pink, a gelatin-encapsuled astaxanthin, on growth parameters, lipid composition, pigment concentration and skin color of commercial size red porgy. Three isonitrogenous diets were formulated containing different levels of astaxantin: Ctrl diet (no pigment added), A50 diet (with 50 mg/kg of astaxanthin) and A80 diet (with 80 mg/ kg of astaxanthin). Four groups of fish (386.29 ± 29.50 g initial weight) were grown in triplicate tanks (10 fish per tank). Ctrl, A50 and A80 groups were fed the respective diets to apparent satiation for 180 days, while a fourth group was fed A50 diet for 90 days followed by A80 diet for another 90 days (A50A80). At the end of trial muscle samples were collected for composition analysis, liver and mesenteric fat for lipid composition determination and skin samples for carotenoids quantification. Dietary astaxanthin supplementation had no effect on growth performance and hepatosomatic index. However, A50A80 group displayed a lower muscle protein content and higher fat content when compared to the other groups. There was no clear effect of dietary astaxanthin supplementation on the liver’s and mesenteric fatty acid profile. Although there was no significant effect on skin total carotenoids, the astaxanthin content tended to increase in fish fed astaxanthin supplemented diets, which was reflected on redness values. Accordingly, dietary astaxanthin supplementation (at 50 or 80 mg/kg for 6 months or at 50 mg/kg for 3 months followed by 80 mg/kg for 3 months) influenced positively skin hue and chroma of red porgy on both pectoral and caudal areas, improving the skin color in commercial sized red porgy and achieving hue and chroma values close to those previously reported for wild individuals. The results reported here provide evidence of the applicability of Carophyll® Pink to improve skin color of large size commercial red porgy, but further studies are needed to optimize this carotenoid supplementation in production farming.