Carotenogenesis and physico-chemical characteristics during maturation of red fleshed papaya fruit (Carica papaya L.)

Carotenoid development of red fleshed papaya fruit (Carica papaya L.) was investigated in the course of a complete pre- and postharvest period using HPLC-DAD coupled to mass spectrometry. Esterified xanthophylls such as ??-cryptoxanthin laurate and caprate were the most abundant pigments during incipient carotenoid biosynthesis. Subsequent fruit maturation led to a gradual accumulation of carotenoids, whereas particularly ??-cryptoxanthin laurate and total lycopene contents disproportionately increased, reaching maximum contents of up to 775 and 3168 ??g/100 g of fresh weight (FW), respectively. Total carotenoid contents of fully ripe papaya ranged from 5414 to 6214 ??g/100 g of FW, while corresponding biosynthetic precursors like phytoene, phytofluene, and ??-carotene were only detected in trace amounts. Due to high contents of vitamin A precursors like ??-carotene and ??-cryptoxanthins, edible parts of the ripe fruit contained 132-166 ??g retinol equivalents per 100 g of FW.Furthermore, the development of morphological and physico-chemical fruit traits was characterized revealing significant correlations to carotenoid accumulation. A ripening index derived from several parameters was developed to easily allow exact assignment of ripening stages in studies of carotenoid development during fruit ontogenesis. Additionally, on-tree ripened versus postharvest ripened fruits were compared revealing striking similarity of their physico-chemical parameters and contents of individual carotenoids.     

Carotenoid composition from the Brazilian tropical fruit camu–camu (Myrciaria dubia)

Camu–camu (Myrciaria dubia) is a small berry, native to the Amazon, known as a rich source of ascorbic acid. The carotenoid composition of this fruit was determined using high performance liquid chromatography-diode array detection on C₁₈ and C₃₀ columns. Fruits produced in two different regions of São Paulo State, Iguape and Mirandópolis, were analysed. All-trans-lutein was the major carotenoid in camu–camu fruits from both regions, ranging from 45% to 55% of the total carotenoid content (160.5 ± 93.1 μg/100 g for Iguape and 601.9 ± 75.6 μg/100 g for Mirandópolis fruits), followed by β-carotene, violaxanthin and luteoxanthin. The levels of lutein, β-carotene, violaxanthin, luteoxanthin and other minor carotenoids were significantly higher in the camu–camu produced in Mirandópolis region, most probably due to the higher temperature and light exposure found in this region, in comparison to those from Iguape. Maturation was also an important feature affecting batches from the same region.     

Expression of carotenoid biosynthetic pathway genes and changes in carotenoids during ripening in tomato (Lycopersicon esculentum)

To study the expression pattern of carotenoid biosynthetic pathway genes, changes in their expression at different stages of maturity in tomato fruit (cv. Arka Ahuti) were investigated. The genes regulating carotenoid production were quantified by a dot blot method using a DIG (dioxigenin) labelling and detection kit. The results revealed that there was an increase in the levels of upstream genes of the carotenoid biosynthetic pathway such as 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR), 4-hydroxy-3-methyl-but-2-enyl diphosphate reductase (Lyt B), phytoene synthase (PSY), phytoene desaturase (PDS) and ζ-carotene desaturase (ZDS) by 2-4 fold at the breaker stage as compared to leaf. The lycopene and β-carotene content was analyzed by HPLC at different stages of maturity. The lycopene (15.33 ± 0.24 mg per 100 g) and β-carotene (10.37 ± 0.46 mg per 100 g) content were found to be highest at 5 days post-breaker and 10 days post-breaker stage, respectively. The lycopene accumulation pattern also coincided with the color values at different stages of maturity. These studies may provide insight into devising gene-based strategies for enhancing carotenoid accumulation in tomato fruits.     

Determination of Carotenoids in Fruits of Rosa sp. (Rosa Canina and Rosa Rugosa) and of Chokeberry (Aronia Melanocarpa)

Carotenoid composition of fruits of two Rosa species (Rosa canina and Rosa rugosa) and of chokeberry (Aronia melanocarpa) was studied by high performance liquid chromatography. Nine carotenoids were determined: three carotenes (lycopene, ζ-carotene, β-carotene) and six xanthophylls (neoxanthin, trans-violaxanthin, cis-violaxan-thin, 5,6-epoxylutein, lutein, β-cryptoxanthin). This high number of compounds classified these fruits among those with the greatest variety of carotenoid pigments. Quantitatively large differences occurred in the carotenoid composition of the three fruits. Rosa hips contained the highest concentrations of total carotenoids, which were mainly comprised of lycopene and βcarotene. Conversely, total xanthophylls were low. In contrast to Rosa, fruits of Aronia were lower in total carotenoids while xanthophylls composed a higher proportion.     

Analysis of carotenoids with emphasis on 9-cis β-carotene in vegetables and fruits commonly consumed in Israel

Recent epidemiological studies have directed the attention from the synthetic all-trans β-carotene to natural carotenoids predominant in fruits and vegetables as possible active ingredients for prevention of cancer and cardiovascular diseases. Seventeen fruits and 17 vegetables commonly consumed in Israel and the β-car-otene-rich alga, Dunaliella bardawil, were analysed for their content of carotenoids with emphasis on 9-cis β-carotene by reversed-phase, 3D photodiode array HPLC. Fourteen carotenoids were eluted in order of decreasing polarity, from polar oxycarotenoids to lipophilic hydrocarbons, and quantified in μg carotenoid per gram freeze-dried plant sample. The richest sources of total carotenoids (>100μg/g dry weight) in Israeli fruits were pittango, mango and papaya while, in vegetables, the predominant types were carrot, dill, parsley, tomato, lettuce, sweet potato and red pepper. Red fruits and vegetables contained mainly lycopene. Yellow and orange fruits and vegetables had high contents of hydrocarbon carotenes with substantial levels of cryptoxanthins and xanthophylls. The green vegetables had high contents of both xanthophylls and hydrocarbon carotenes. Relatively high ratios (9-cis to all-trans β-carotene) of above 0.2 g/g were noted in sweet potato, papaya, parsley, lettuce, dill, apricot, pepper, prune and pumpkin, compared to the high ratio of 9-cis to all-trans β-carotene in the alga Dunaliella (~ 1.0 g/g). The high content of 9-cis β-carotene in certain fruits and vegetables and the wide variety of carotenoids and stereoisomers of carotenoids in all plants should shift nutritional and medical attention from the synthetic all-trans β-carotene toward natural carotenoids as potential candidates for chemoprevention.     

Comparative fruit colouration in watermelon and tomato

The characteristic red pigmentation of watermelon and tomato fruits is determined by accumulation of the carotenoid pigment lycopene and this phenotype is polyphyletic. Since several carotenoids are known to have health promoting activity, and watermelon can be a significant source of lycopene and other carotenoids, it is important to understand the genetic basis of watermelon fruit-specific carotenoid biosynthesis. Unlike tomato, very little is known about the regulation of carotenoid biosynthesis during fruit development in watermelon, a non-climacteric fruit. We have HPLC analyzed the carotenoids of red, yellow and orange watermelons and compared their carotenoid patterns with those of known fruit colour mutants of tomato. Interestingly, we could detect tomato mutant equivalents to most watermelon fruit colour phenotypes, including r, og, B and t.     

Analysis of a complex mixture of carotenes from oil palm (Elaeis guineensis) fruit extract

A carotene extract from the fruits of the oil palm (Elaeis guineensis) was analysed by HPLC employing a C₃₀ column for better separation efficiency. A multitude of cis-isomers of α-, β- and γ-carotene were separated. Detailed assignment was possible by subjecting pure standards of α-, β- and γ-carotene to isomerisation and comparing spectral data and order of elution to literature data. α- and β-carotene were found to be the most abundant carotenoids comprising 12.3% and 17.9%, respectively, of a (roughly) 30% oil suspension of oil palm carotenes in vegetable oil. A large proportion (about 40%) of α- and β-carotene was in the form of cis-isomers. The γ-carotene content was found to be 0.38% and other carotenes like phytoene, phytofluene, ζ-carotene, lycopene and possibly β-zeacarotene were found as well but were not quantified.     

Identification of key enzymes involved in the accumulation of carotenoids during fruit ripening of Lycium barbarum L. by a proteomic approach

The isobaric tag for relative and absolute quantification (iTRAQ)-based proteomic approach was used to identify the key enzymes involved in the carotenoids accumulation from flowering to maturity stage of Lycium barbarum L. A total of 1,799 differentially expressed proteins were identified, and 190 were related to carotenoids metabolism. Six enzymes, including phytoene synthase, phytoene desaturase, zeta-carotene desaturase, P450 carotenoid beta-ring hydroxylase (LUT5), neoflavin synthase and nine-cis-epoxycarotenoid dioxygenase were found to be associated with the carotenoid accumulation during L. barbarum ripening by the proteomic analysis and qRT-PCR. The changes in the chromaticity and carotenoid contents during fruit ripening were also studied. The contents of carotenoid, except violaxanthin were strongly correlated with the expression levels of the key enzymes, and the colours were strongly correlated with the contents of carotenoid. This study may contribute to further investigations of the accumulation mechanism of carotenoids in L. barbarum.     

In vitro bioaccessibility of carotenoids and tocopherols from fruits and vegetables

Aim of the study

To assess the in vitro bioaccessibility of carotenoids, including xanthophyll esters, and tocopherols from fruits and vegetables.

Results

Stability for carotenoids and tocopherols was over 70%. Xanthophyll esters were cleaved by cholesterol esterase but not by human pancreatic lipase. Less than 40% of the β-cryptoxanthin initially present was hydrolyzed and the amount of free xanthophylls recovered was higher when liquid was used than when fresh homogenized matrix was employed. cis-Isomers of β-carotene and lutein did not significantly increase during the process. Xanthophylls were more efficiently transferred into supernatants than tocopherols and β-carotene. cis-Carotenoids, epoxy-xanthophylls and ester forms were also transferred.

Conclusion

The results are consistent with observations in other in vitro digestion models and human studies and support the usefulness of in vitro assessment to study food-related determinants of the bioavailability of carotenoids and tocopherols from fruits and vegetables.     

The content of polyphenols and carotenoids in three apricot cultivars depending on stage of maturity and geographical region

During the ripening of three apricot cultivars (“Keckemetska ruza”, “Madjarska najbolja” and “Velika rana”) grown in two different geographical region of Croatia the changes of polyphenols and carotenoids were determined by using high-performance liquid chromatography (HPLC) with UV–Vis photo diode array detection. The content of individual polyphenols during ripening was quite similar, whereas their amount differed significantly. Immature fruits showed the highest level of polyphenols, which decreased at semi-mature fruits and did not change remarkably in commercial mature fruits. Among polyphenols, flavan-3-ols, chlorogenic acid and quercetin-3-rutinoside were dominant in all ripening stages of all apricot cultivars. The quantity of polyphenols during apricot fruits ripening depended on cultivars, while the region of cultivation did not have remarkable influence on polyphenols amount. During ripening carotenoids increased significantly especially β-carotene which represented 70–85% of the total carotenoid content. Besides β-carotene, in all apricot cultivars γ-carotene was found. α-Carotene, zeaxantin and lutein were found in cv. “Madjarska najbolja” and in cv. “Velika rana” α-carotene and lutein were determined. The region of cultivation and the cultivar were important factors, which influenced the carotenoid amount of apricot fruits, and this content was higher in cultivars grown in Mediterranean region.     

Phytochemical and antioxidant characterization of mamey (Pouteria sapota Jacq. H.E. Moore & Stearn) fruit

Phytochemical compounds in fruits and vegetables have gained great importance in the last few years because of the increasing evidence suggesting their antioxidant and prevention of chronic diseases. Carotenoids, phenolics, flavonoids, and vitamins E and C, are among these phytochemicals. Several fruits have been characterized so far for their antioxidant and health properties but there is still limited information on fruits from the tropic. Therefore, the objective of this study was the characterization of mamey fruit (Pouteria sapota Jacq. H. E. Moore & Stearn) with regard to their antioxidant capacity and phytochemical profile. Phenolics, carotenoids and ??-tocopherol were quantified and identified by HPLC-DAD-Mass Spectrometry (LC-MS), and DPPH and FRAP assays were used to evaluate antioxidant capacity. Hydrophilic extracts of mamey fruit showed higher antioxidant capacity than the lipophilic portion. Total soluble phenols content was 28.5 mg GAE/100 g fw, being p-hydroxybenzoic acid as the main phenolic that was identified. Total carotenoid content was 1127.9 ??g ??-carotene/100 g fw with ??-carotene being the main contributor, in addition to lutein, and violoxanthin. Concentration of ??-tocopherol was 360.0 ??g/100 g fw. Results of this study suggest that mamey fruit is a good source of carotenoids and its inclusion in the diet is recommended.     

Carotenoid composition and vitamin A value of the brasilian fruit Cyphomandra betacea

The carotenoid pigments of the fruit of the Tree Tomato Cyphomandra betacea were identified and quantified. β-carotene, β-cryptoxanthin, ζ-carotene, 5,6-monoepoxy-β-carotene, lutein and zeaxanthin were detected in both the pulp and the peel. The quantitative patterns of the pulp and the peel were similar, with cryptoxanthin and β-carotene predominating. The high average vitamin A value (2475 IU/100 g edible portion) is due to the principal carotenoids that have provitamin A activity.     

Phytochemical and antioxidant characterization of the fruit of black sapote (Diospyros digyna Jacq.)

Consumption of fruits and vegetables has been associated with a healthy state which has been attributed, in part, to their antioxidant capacity. Characterization of the bioactive compounds with antioxidant activity found in fruits cultivated in the tropic is limited. Thus, the objective of the present work was the characterization of phytochemicals and antioxidants of the fruit of black sapote (Diospyros digyna Jacq.). HPLC-DAD-Mass Spectrometry (HPLC-MS) analyses were used to identify and quantify phenolics, carotenoids and tocopherols. Total soluble phenolic content was 247.8 mg GAE/100 g fw (fresh weight). Important phenolics identified were sinapic acid, myricetin, ferulic acid, and catechin. Total carotenoid content was 399.4 ??g of ??-carotene/100 g fw, and ??-carotene and lutein were the main carotenoids identified. ??-Tocopherol concentration was 672.0 ??g/100 g dry weight. Antioxidant capacity as measured by the DPPH and FRAP assays was higher in the hydrophilic than in the lipophilic extract, and it is thought to be due mainly to the phenolic content of this fruit. Results suggest that the fruit of black sapote has an antioxidant capacity comparable to other important fruits, and its inclusion in the diet is therefore recommended.     

Comparative study of the effect of the maturation process of the olive fruit on the chlorophyll and carotenoid fractions of drupes and virgin oils from Arbequina and Farga cultivars

Two varieties of olive fruit (Arbequina and Farga) have been characterized by their contents and types of chlorophyll and carotenoid pigments during the olive ripening. Independently of the initial content, the pigment concentrations decreased with fruit maturity. Chlorophyllides a and b, esterified xanthophylls and α-carotene were only detected in Arbequina fruits. Moreover, the synthesis of esterified xanthophylls, when the skin fruit colour changed from green to turning-colour only, was observed in this variety. The pigments transferred from fruit to the oil were also studied. All the pigments that were found in the fruit were transferred to the oils, in addition to derivative pigments associated with acidic medium in the oil extraction process. The destruction of the chlorophyll fraction was greater than of the yellow pigments during the olive oil extraction process.     

Influence of extraction with ethanol or ethyl acetate on the yield of lycopene, β-carotene,phytoene and phytofluene from tomato peel powder

This study evaluated the extraction yield of the food grade solvents ethanol and ethyl acetate by extracting lycopene, β-carotene, phytoene and phytofluene from tomato peel powder at varying heating intensities, and the influence of the solvent and heating intensity on carotenoids isomerization and degradation during extraction. The heat treatments assayed were 25, 35, 50 and 60 °C which were applied for periods of 5, 10, 20, 30 or 40 min. The carotenoid yield was higher in the extractions with ethanol than with ethyl acetate. In general, the temperature increase caused an increase in the carotenoid concentrations; however in the extractions performed with ethanol at 60 °C, the yield of (all-E)-lycopene and their (Z)-isomers was lower than at 50 °C. This could indicate that a great isomerization is produced in the high temperature extractions with ethanol but the oxidative degradation is the predominant reaction. On the contrary, the obtained results in the extractions with ethyl acetate indicate that the isomerization is the predominant reaction.     

Carotenoids and antioxidant capacities from Canarium odontophyllum Miq. fruit

Carotenoids were isolated and identified from peel, pulp and seed fractions of Canarium odontophyllum Miq., and their antioxidant capacities were evaluated. all-trans-β-carotene was present in a large amount in peel (69.5 ± 1.0 mg/kg), followed by pulp (31.1 ± 0.76 mg/kg) and seed (15.1 ± 3.0 mg/kg). Additionally, 15-cis-β-carotene, 9-cis-β-carotene and 13-cis-β-carotenes were also major contributors to carotenoid contents in peel, pulp and seed fractions. Pulp exhibited excellent β-carotene bleaching activity, significantly higher than peel and seed; high 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity, whereas peel exhibited significantly higher scavenging activity of 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals. All the extracts exhibited good inhibitory effect against hydrogen peroxide-induced haemoglobin oxidation, ranging from 45.3 to 59.7%. This is the first report about carotenoids and antioxidant capacities from C. odontophyllum fruit, and indicates that this fruit can be explored and promoted as a potential source of natural antioxidants.     

Ultra-performance liquid chromatographic separation of geometric isomers of carotenoids and antioxidant activities of 20 tomato cultivars and breeding lines

All-trans-lutein, lycopene, β-carotene and their 22 cis-isomers in 20 tomato breeding were separated and identified by a rapid and sensitive UPLC method using a 1.7 μm C₁₈ column and a new gradient mobile phase based on methanol-MTBE-water in 15 min. All-trans-carotenoids were predominant, but 9-cis, 13-cis-lutein, 5-cis, 9-cis, 13-cis, 15-cis, di-cis-lycopene, 9-cis, 13-cis, 15-cis and di-cis-β-carotene were also found. The cis-isomers were identified using absorption around 330 nm and the Q-ratio. The total antioxidant activities as evaluated by PCL and DPPH assays were found to correlate well with the total carotenoid content, but not with the individual carotenoid or its different isomers. This paper provides an efficient analytical method for obtaining a complete picture of carotenoids in tomatoes. It can be a valuable tool for plant breeders, food processors and researchers in developing designer tomatoes and tomato-products with unique carotenoid compositions, and functional properties.     

Enzymatic carotenoid degradation and aroma formation in nectarines (Prunus persica)

C-13 norisoprenoids such as β-ionone, 3 hydroxy-β-ionone, and 3 hydroxy-5,6-epoxy-β-ionone are important carotenoid related aroma compounds in fruits.Apart from chemical degradation (photo-oxygenation, (auto)oxidation, thermal degradation), enzymatic cleavage is an important flavor formation pathway [Enzell, C. (1985). Biodegradation of carotenoids – an important route to aroma compounds. Pure and Applied Chemistry, 57(5), 693–700; Winterhalter, P., & Rouseff, R. (Eds.). (2001). Carotenoid-derived aroma compounds. ACS symposium series 802, Washington, DC: American Chemical Society (ISBN 0-8412-3729-8)]. In nectarines, the content of carotenoid degradation products depends on the degree of maturity. In fully ripened fruit up to 40% of the aglycons are C-13 norisoprenoids [Aubert, C., Günata, Z., Ambid, C., & Baumes, R. (2003). Changes in physicochemical characteristics and volatile constituents of yellow- and white-fleshed nectarines during maturation and artificial ripening. Journal of Agriculture and Food Chemistry, 51(10), 3083–3091]. Regiospecific carotenoid cleavage enzymes have been shown to be involved in aroma formation in different plants and fruits [Bouvier, F., Suire, C., Mutterer, J., & Camara, B. (2003). Oxidative remodeling of chromoplast carotenoids: identification of the carotenoid dioxygenase CsCCD and CsZCD genes involved in crocus secondary metabolite biogenesis. Plant Cell, 15(1), 47–62; Fleischmann, P., Studer, K., & Winterhalter, P., (2002). Partial purification and kinetic characterization of a carotenoid cleavage enzyme from quince fruit (Cydonia oblonga). Journal of Agriculture and Food Chemistry, 50(6), 1677–1680; Fleischmann, P., Watanabe, N., & Winterhalter, P., (2003). Enzymatic carotenoid cleavage in star fruit (Averrhoa carambola). Phytochemistry, 63(2), 131–137; Schwartz, Qin & Zeevaart, 2001].This paper describes the isolation and partial characterization of carotenoid cleavage enzymes in nectarines, isolated from ripe skin of commercially available fruit. The enzyme is characterized by its kinetic parameters (v_max, K_m, time constant, temperature dependence, activation energy).     

Carotenoid composition and retinol equivalent in plants of nutritional and medicinal importance: Efficacy of β-carotene from Chenopodiumalbum in retinol-deficient rats

This study aimed to (1) quantify carotenoids in leafy vegetables and plants of nutritional and medicinal importance, (2) evaluate retinol equivalent (RE) of provitamin-A carotenoids and (3) determine efficacy of β-carotene from Chenopodium album and to compare with retinol formed on feeding to retinol-deficient rats for 3 weeks. β-Carotene and lutein contents (mg/100 g dry weight) ranged from 1.5 to 120 and 11.7 to 185 (leafy greens) and 0.4 to 34.7 and 11.8 to 679 (medicinal plants) whereas, α-carotene ranged from 0.3 to 35.6 (leafy greens) and 0.1 to 15.7 (medicinal plants). RE values (mg%) ranged from 0.4 to 20 and 0.42 to 5.8 in leafy greens and medicinal plants. Efficacy of β-carotene (2400 μg/kg diet) from C. album in retinol-deficient rats revealed a 93.6% rise in plasma retinol levels from 0.53 to 8.4 μM. The plants analysed are a good source of retinol precursors and biologically active lutein; therefore can be exploited to meet carotenoid requirements.