HPLC-PDA-MS/MS of anthocyanins and carotenoids from dovyalis and tamarillo fruits

Anthocyanins and carotenoids are natural pigments responsible for the color of vegetables and fruits, and they are also bioactive compounds, both demonstrating important biological, therapeutic, and preventative properties. Considering the biodiversity of edible fruits, high performance liquid chromatography coupled to photodiode array and mass spectrometry detectors (HPLC-PDA-MS) was used to establish the composition of carotenoids and anthocyanins from dovyalis and tamarillo fruits. Ten anthocyanins and 26 carotenoids were found in dovyalis, whereas tamarillo showed 3 anthocyanins and 17 carotenoids. Higher contents of anthocyanins and carotenoids were found in dovyalis, 42.0 and 6.6 mg/100 g, respectively, as compared to tamarillo fruits with 8.5 and 4.4 mg/100 g, respectively. Although these fruits belong to different families, delphinidin 3-rutinoside and beta-cryptoxanthin were found to be, respectively, the major anthocyanin and carotenoid in both fruits.     

Change in carotenoids and antioxidant vitamins in tomato as a function of varietal and technological factors

The change in the carotenoid and bioantioxidant content of tomato as a function of varietal and technological factors was investigated in the present work. No great differences were found between cultivars for fresh consumption (salad tomatoes) and those for processing in ascorbic acid content. The concentration of ascorbic acid ranged between 14.6 and 21.7 mg/100 g fresh weight of ripe tomato fruit. Processing cultivars contained higher amounts of tocopherols, particularly alpha-tocopherol than tomatoes for fresh consumption. Significant differences could be obtained between the examined varieties with regard to carotenoid concentration. The different tomatoes varied not only in the total carotenoid content but also in the qualitative distribution of some pigments such as lycopene, beta-carotene and lutein. During heat-based processing, ascorbic acid, tocopherols, and carotenoids showed different role and response. Ascorbic acid, alpha-tocopherol quinone, and beta-carotene were the most susceptible components toward thermal degradation.     

Influence of combined biotic and abiotic stress on nutritional quality parameters in tomato (Solanum lycopersicum)

Induction of abiotic stress in tomato plants has been proposed as a mechanism for improving the nutritional quality of fruits. However, the occurrence of biotic stress can interfere with normal abiotic stress responses. In this study, the combined effect of water stress and infection with plant-parasitic nematodes on the nutritional quality of tomato was investigated. Plants were exposed to one or both stresses, and the levels of phenolic compounds, carotenoids, and sugars in fruits were analyzed as well as physiological responses. Levels of carotenoids lycopene and β-carotene were lower in water-stressed tomatoes but exhibited a different response pattern under combined stress. Nematode stress was associated with increased flavonoid levels, albeit with reduced yields, while chlorogenic acid was increased by nematodes, water stress, and the combined stress. Sugar levels were higher only in tomatoes exposed to both stresses. These results emphasize the importance of studying plant stress factors in combination.     

Susceptibility of the tomato mutant high pigment-2dg (hp-2dg) to Orobanche spp. infection

The consumption of natural products with potential health benefits has been continuously growing, and enhanced pigmentation is of major economic importance in fruits and vegetables. The tomato hp-2 ( dg ) is an important mutant line that has been introgressed into commercial tomato cultivars marketed as lycopene rich tomatoes (LRT) because of their enhanced fruit pigmentation, attributed to higher levels of carotenoids, including lycopene. Strigolactones are signaling compounds that mediate host finding in root parasitic plants and are biosynthetically derived from carotenoids. Considering the high carotenoid content of the hp-2 ( dg ) mutant, we studied its susceptibility to the root parasite Orobanche. In a field experiment, the average number of Orobanche aegyptiaca plants growing on hp-2 ( dg ) was surprisingly significantly reduced compared with its isogenic wild-type counterpart. In vitro assays and LC-MS/MS analysis showed that this reduction was associated with a lower production of strigolactones, which apparently renders the high-carotenoid hp-2 ( dg ) mutant less susceptible to Orobanche.     

Characterization of phytochemicals and antioxidant activities of a purple tomato (Solanum lycopersicum L.)

A newly developed nongenetically modified purple tomato V118 was investigated for its phytochemical compositions and antioxidant activities. A highly efficient and sensitive UPLC method was developed for both the phenolics and carotenoids, which showed that in addition to the phytochemicals commonly known for tomatoes, V118 had a unique composition of anthocyanins. The total carotenoid content of V118 was 234.78 μg/g dry weight (DW), and the total phenolic content was 659.11 mg GAE/100 g DW. The antioxidant activities of the lipophilic extract as measured by the PCL and ORAC-L assays were 30.11 μmol TE/g DW and 11.97 μmol TE/g DW, respectively, while the hydrophilic extracts as determined by the ORAC-H and FRAP assays were 323.23 μmol TE/g DW and 54.95 μmol AAE/g DW, respectively. The LC-MS study showed three major anthocyanins, which were mainly acylglycosides of petunidin and malvidin. This study showed that purple tomatoes such as V118 possess additional phytochemicals like anthocyanins, which can potentially have added health benefits.     

Carotenoid pigmentation affects the volatile composition of tomato and watermelon fruits, as revealed by comparative genetic analyses

Tomato near-isogenic lines differing in fruit carotenogenesis genes accumulated different aroma volatiles, in a strikingly similar fashion as compared to watermelon cultivars differing in fruit color. The major volatile norisoprenoids present in lycopene-containing tomatoes and watermelons were noncyclic, such as geranial, neral, 6-methyl-5-hepten-2-one, 2,6-dimethylhept-5-1-al, 2,3-epoxygeranial, (E,E)-pseudoionone, geranyl acetone, and farnesyl acetone, seemingly derived from lycopene and other noncyclic tetraterpenoids. Beta-ionone, dihydroactinodiolide, and beta-cyclocitral were prominent in both tomato and watermelon fruits containing beta-carotene. Alpha-ionone was detected only in an orange-fleshed tomato mutant that accumulates delta-carotene. A yellow flesh (r) mutant tomato bearing a nonfunctional psy1 gene and the yellow-fleshed watermelon Early Moonbeam, almost devoid of carotenoid fruit pigments, also lacked norisoprenoid derivatives and geranial. This study provides evidence, based on comparative genetics, that carotenoid pigmentation patterns have profound effects on the norisoprene and monoterpene aroma volatile compositions of tomato and watermelon and that in these fruits geranial (trans-citral) is apparently derived from lycopene in vivo.     

Processing effects on lycopene content and antioxidant activity of tomatoes

Consumption of tomato products has been associated with decreased risk of some cancer types, and the tomato antioxidant, lycopene, is thought to play an important role in the observed health effects. In this study, four carotenoids, trans-lycopene, phytofluene, phytoene, and zeta-carotene, were quantified in tomato products. Samples of raw tomatoes, tomato juice after hot break scalder, and final paste were obtained from two different processing plants over two years. Comparison of carotenoid levels throughout processing indicated that lycopene losses during processing of tomatoes into final paste (25-30 degrees Brix) ranged from 9 to 28%. The initial Brix level of the raw tomatoes appeared to influence the amount of lycopene loss that occurred, possibly due to the differences in processing time required to achieve the final desired Brix level of the paste. In general, no consistent changes in the other carotenoids were observed as a function of processing. The antioxidant activity of fresh tomatoes, tomato paste, and three fractions obtained from these products (i.e., aqueous, methanol, and hexane fractions) was also determined. In both a free radical quenching assay and a singlet oxygen quenching assay, significant antioxidant activity was found in both the hexane fraction (containing lycopene) and the methanol fraction, which contained the phenolic antioxidants caffeic and chlorogenic acid. The results suggest that in addition to lycopene, polyphenols in tomatoes may also be important in conferring protective antioxidative effects.     

Influence of organic versus conventional agricultural practice on the antioxidant microconstituent content of tomatoes and derived purees; consequences on antioxidant plasma status in humans

The present study aims first to compare the antioxidant microconstituent contents between organically and conventionally grown tomatoes and, second, to evaluate whether the consumption of purees made of these tomatoes can differently affect the plasma levels of antioxidant microconstituents in humans. When results were expressed as fresh matter, organic tomatoes had higher vitamin C, carotenoids, and polyphenol contents (except for chlorogenic acid) than conventional tomatoes. When results were expressed as dry matter, no significant difference was found for lycopene and naringenin. In tomato purees, no difference in carotenoid content was found between the two modes of culture, whereas the concentrations of vitamin C and polyphenols remained higher in purees made out of organic tomatoes. For the nutritional intervention, no significant difference (after 3 weeks of consumption of 96 g/day of tomato puree) was found between the two purees with regard to their ability to affect the plasma levels of the two major antioxidants, vitamin C and lycopene.     

Profiling of carotenoids in tomato juice by one- and two-dimensional NMR

Epidemiological data have shown a link between dietary intake of tomatoes and tomato products (rich in carotenoids) and a decreased risk of chronic diseases. The carotenoid profile in tomato products depends on tomato variety as well as the thermal conditions used in processing. The final carotenoid profile may affect the bioaccessibility and bioavailability of these biomolecules. Therefore, nondestructive, reliable methods are needed to characterize the structural and stereochemical variation of carotenoids. CDCl(3) rapid extraction was used to extract carotenoids from tomato juice as an alternative rapid procedure that minimizes solvents and time consumption prior to NMR analysis. The profile of these biomolecules was characterized by application of high-resolution multidimensional NMR techniques using a cryogenic probe. The combination of homonuclear and heteronuclear two-dimensional NMR techniques served to identify (all-E)-, (5Z)-, (9Z)-, and (13Z)-lycopene isomers and other carotenoids such as (all-E)-beta-carotene and (15Z)-phytoene dissolved in the extracted lipid mixture. The use of one-dimensional NMR enabled the rapid identification of lycopene isomers, thereby minimizing further isomerization of (all-E)-lycopene as compared to HPLC data. On the basis of the assignments accomplished, the carotenoid profile of typical tomato juice was successfully determined with minimal purification procedures.     

Antioxidative activity and carotenoid and tomatine contents in different typologies of fresh consumption tomatoes

The phytonutrient intake associated with tomato consumption depends also on cultivar and fruit ripening stage. This work associates the antioxidative ability, the level of carotenoids, and the amount of glycoalkaloids to the main carpometric characteristics of four different typologies of tomatoes: "cherry", "cluster", "elongated," and "salad". These typologies have different weights and shapes, and they are usually consumed in the Mediterranean area at different ripening stages. Results showed that the considered tomato typologies also differ in their antioxidative ability and their carotenoid and glycoalkaloid contents. Growing conditions are also important in determining fruit characteristics: the analysis of the same cultivar of cherry tomato produced under the influence of moderate salt stress showed increases in the lipophilic antioxidative ability and the amount of carotenoid, whereas the level of glycoalkaloid decreased.     

Changes in carotenoid and ascorbic acid contents in fruits of different tomato genotypes related to the depletion of UV-B radiation

The aim of the present study was to investigate if the depletion of UV-B radiation affected the most representative carotenoids as well as the ascorbic acid content in tomato fruits, harvested at both breaker and firm red stages. To do this, three tomato genotypes, DRW 5981, HP 1, and Esperanza, were grown inside a greenhouse either covered with polyethylene transparent to UV-B or depleted of UV-B by a special covering film. The antioxidant properties of the fruits were evaluated on the water-insoluble fractions according to the ABTS method. UV-B effect on antioxidant activity was negligible in DRW and HP 1 genotypes, whereas it was detrimental in Esperanza at both ripening stages. This genotype seems to have a negligible capability of accumulating carotenoids and a great susceptibility to detrimental effects of UV-B; conversely, the DRW genotype shows high carotenoid levels under sunlight conditions and a further promotion by UV-B. On the other hand, the HP 1 mutant displays an intermediate behavior and represents the only genotype favored by UV-B with respect to ascorbic acid accumulation.     

Animal-based organic nutrition induces comparable fruit quality to that of inorganic fertigation in soilless-grown grape tomato

The aim of the present study was to determine the effect of animal-based organic nutrition and environmental parameters on tomato fruit quality, as well as to establish relations among colour and morphological values performed by the Tomato Analyzer (TA) software application. Organic tomato fruits produced by three organic nutrient solutions, which consisted of different mixtures of several OMRI certified nitrogen fertilizers and one inorganic nutrient solution (Steiner’s solution) as the control, were evaluated for their polyphenol and carotenoid content. We used Tomato Analyzer (TA) to evaluate fruit size and shape. Moreover, we implemented a digital image analysis tool, Color Test (CT), as part of the TA software application to collect and analyse fruit colour parameters. The application of organic fertilizers positively affected the total hydrolysable and condensed polyphenols of tomato fruits compared to the control. The high air temperature (>30°C) and sub-optimal light intensity negatively affected the carotene content of tomato fruits, as well as their morphological and colour attributes. Plants fed with organic solutions containing ASA + ASB + NK2SO4 showed comparable morphology and fruit colour attributes to those of the control plants that received Steiner’s nutrient solution. The results indicated that the application of organic fertilizers positively affected the total hydrolysable and condensed polyphenols of tomato fruits compared to the control. Plants fed with the Steiner’s nutrient solution exhibited the highest carotenoid content in tomato fruits. Organically produced tomatoes through animal-based fertilizer application displayed similar fruit morphology and colour attributes compared to conventionally produced tomatoes.     

Carotenoids in white- and red-fleshed loquat fruits

Fruits of 23 loquat ( Eriobotrya japonica Lindl.) cultivars, of which 11 were white-fleshed and 12 red-fleshed, were analyzed for color, carotenoid content, and vitamin A values. Color differences between two loquat groups were observed in the peel as well as in the flesh. beta-Carotene and lutein were the major carotenoids in the peel, which accounted for about 60% of the total colored carotenoids in both red- and white-fleshed cultivars. beta-Cryptoxanthin and, in some red-fleshed cultivars, beta-carotene were the most abundant carotenoids in the flesh, and in total, they accounted for over half of the colored carotenoids. Neoxanthin, violaxanthin, luteoxanthin, 9- cis-violaxanthin, phytoene, phytofluene, and zeta-carotene were also identified, while zeaxanthin, alpha-carotene, and lycopene were undetectable. Xanthophylls were highly esterified. On average, 1.3- and 10.8-fold higher levels of colored carotenoids were observed in the peel and flesh tissue of red-fleshed cultivars, respectively. The percentage of beta-carotene among colored carotenoids was higher in both the peel and the flesh of red-fleshed cultivars. Correlations between the levels of total colored carotenoids and the color indices were analyzed. The a* and the ratio of a*/ b* were positively correlated with the total content of colored carotenoids, while L*, b*, and H degrees correlated negatively. Vitamin A values, as retinol equivalents (RE), of loquat flesh were 0.49 and 8.77 microg/g DW (8.46 and 136.41 microg/100 g FW) on average for white- and red-fleshed cultivars, respectively. The RE values for the red-fleshed fruits were higher than fruits such as mango, red watermelon, papaya, and orange as reported in the literature, suggesting that loquat is an excellent source of provitamin A.     

彩色油菜花瓣色素成分研究

为明确彩色油菜花瓣色素成分和色素呈色机制,提高彩色油菜育种效率,本研究比较了红色、橙色、黄色和白色4种色系共10种彩色油菜花瓣色素代谢的次生代谢物含量,并采用液质联用仪(HPLC-MS/MS)分析红色、橙色和黄色油菜花瓣中花青素和类胡萝卜素含量,同时对花青素合成关键酶活性和编码关键酶活性的基因表达量进行分析。结果表明,各色系油菜花瓣积累了大量酚类和类黄酮化合物。红色系和橙色系油菜花瓣中含有丰富的原花青素和花色苷;黄色系油菜花瓣中不含原花青素,但含花色苷;白色系油菜花瓣既不合成原花青素,也不合成花色苷。红色和橙色油菜花瓣中的花色苷主要成分是矢车菊素和芍药素。红色、橙色、黄色和白色油菜花瓣中均含有胡萝卜素和番茄红素,且4种花色花瓣中的番茄红素含量均高于胡萝卜素。红色、橙色和黄色油菜花瓣的番茄红素含量仅为对应花色花瓣花青素的0.25%、0.15%、0.02%。基因表达分析结果表明,引起花青素在不同色系油菜花瓣中积累差异的基因为二氢黄酮醇还原酶基因(DFR)和花青素还原酶基因(ANS),在红色、橙色和橙黄色花瓣中DFR和ANS的表达量分别为黄色花瓣的100倍和1 000倍以上。相关性分析表明,彩色油菜DFR和ANS基因的表达量与花色苷含量呈高度正相关性。但是,DFR和ANS活性在红色、橙色、黄色和白色油菜花瓣中差异不显著。本研究为彩色油菜进行基因操纵中靶基因的选择和合理的育种线路提供了理论基础。     

Eelgrass slabs, a soilless culture substrate that inhibits adhesion of fungi and oomycetes and enhances antioxidant activity in tomato

Composed of a marine plant, Zostera sp., eelgrass slabs are a novel organic substrate for soilless cultures used in tomato production. The benefit of using eelgrass slabs for growing tomatoes was assessed by comparing it with coconut fiber slabs in regard to contamination by Pythium spp. and to the antioxidant properties of tomato fruits. First, tomato root contamination by Pythium spp. was studied by direct plate counting, and a molecular comparison of fungal and oomycete communities was conducted using PCR-DHPLC. Second, the antioxidant properties of tomato fruits were analyzed by measuring total phenol and carotenoid contents and by evaluating radical scavenging activity. Compared to plants grown on coconut fiber slabs, those on eelgrass slabs presented a lower rate of Pythium spp. root contamination. Moreover, culture on eelgrass slabs produced fruits with better radical scavenging activity and higher total phenol content compared to controls. Carotenoid content was not affected by the type of substrate. This study highlights the value of detrital leaves of Zostera sp. as a substrate for soilless culture that reduces root contamination and also promotes the production of tomato fruits with better nutritional value.     

Nutritional value of cherry tomatoes (Lycopersicon esculentum Cv. Naomi F1) harvested at different ripening stages

The average content of some classes of antioxidants is generally higher in cherry tomatoes than in normal-sized berries. The aim of this work was to assess the nutritional value of cherry tomato (cv. Naomi F1) by investigating the compositional pattern of berries harvested at different ripening stages and evaluating, in particular, all of the main antioxidants (carotenoids, ascorbic acid, phenolic compounds, and alpha-tocopherol) and the antioxidant activity of the water-soluble and water-insoluble fractions. Results confirmed the relatively high level of carotenoids in cherry tomato but showed that not all biologically active compounds necessarily increase in tomatoes picked at later stages of ripeness. Cherry tomatoes harvested at full ripeness exhibited the highest level of carotenoids and antioxidant activity in the water-insoluble fraction. On the other hand, no significant differences in ascorbic acid content were observed at different ripening stages, whereas the main phenolics content and the antioxidant activity of water-soluble fraction showed slight, but significant, decreases at later stages of ripeness.     

Determination of ascorbic acid and carotenoids in food commodities by liquid chromatography with mass spectrometry detection

Two methods, one to determine ascorbic acid and one to determine lycopene and beta-carotene, in vegetables and fruits by liquid chromatography coupled with mass spectrometry (LC-MS) have been established. The chromatographic separation of the studied compounds and their MS parameters were optimized to improve selectivity and sensitivity. In both methods, separation was carried out with two coupled columns, first a C(18) and then a dC(18), using as mobile phase 70% methanol (0.005% acetic acid) and 30% acetic acid 0.05% for ascorbic acid determination and a mixture of methanol, tetrahydrofuran, and acetonitrile (60:30:10 v/v/v) for carotenoid analysis in isocratic mode. The molecular ion was selected for the quantification in selective ion monitoring (SIM) mode. Ascorbic acid was detected with electrospray ionization probe (ESI) in negative mode, while chemical ionization atmospheric pressure (APCI) in positive mode was used for the target carotenoids. The methodology for ascorbic acid analysis is based on an extraction with polytron using methanol and a mixture of methaphosphoric acid and acetic acid. Extraction of the carotenoids was carried out with tetrahydrofuran/methanol (1:1) (v/v). The proposed methods were applied, after their corresponding validations, to the analysis of four varieties of tomatoes, tomato in tin enriched and dried tomato, and to the analysis of mango and kiwi fruits, to compare the content in these compounds. Moreover, the influence of the process of freezing and the effect that the manipulation/preservation has in the content of ascorbic acid in tomato have also been studied.     

Metabolic Profiling of Glucosinolates, Anthocyanins, Carotenoids, and Other Secondary Metabolites in Kohlrabi (Brassica oleracea var. gongylodes)

We profiled and quantified glucosinolates, anthocyanins, carotenoids, and other secondary metabolites in the skin and flesh of pale green and purple kohlrabis. Analysis of these distinct kohlrabis revealed the presence of 8 glucosinolates, 12 anthocyanins, 2 carotenoids, and 7 phenylpropanoids. Glucosinolate contents varied among the different parts and types of kohlrabi. Glucoerucin contents were 4-fold higher in the flesh of purple kohlrabi than those in the skin. Among the 12 anthocyanins, cyanidin 3-(feruloyl)(sinapoyl) diglucoside-5-glucoside levels were the highest. Carotenoid levels were much higher in the skins than the flesh of both types of kohlrabi. The levels of most phenylpropanoids were higher in purple kohlrabi than in pale green ones. trans-Cinnamic acid content was 12.7-fold higher in the flesh of purple kohlrabi than that in the pale green ones. Thus, the amounts of glucosinolates, anthocyanins, carotenoids, and phenylpropanoids varied widely, and the variations in these compounds between the two types of kohlrabi were significant.     

Changes in chloroplast pigments of olive varieties during fruit ripening

Changes in chlorophyll and carotenoid pigments of five olive (Olea europaea L.) varieties destined for milling were investigated at six consecutive ripening stages. There was a manifest dependence between olive variety, moment of picking, and chloroplast pigment composition of the fruits. Although the content of chlorophylls and carotenoids differed with fruit variety, ripening always involved their gradual loss, which becames more pronounced with increased presence of anthocyanin compounds. The relative rates of disappearance of chlorophylls and carotenoids were markedly different between varieties, implying that the catabolism of these pigments takes place at a relative rate inherent to each variety. The varieties less rich in pigments showed the most extreme behavior. The highest relative rate of disappearance was observed in fruits of the Blanqueta variety, and the lowest was observed in those of Arbequina. The chlorophyll a/chlorophyll b ratio remained practically constant during ripening, with a value very similar for Hojiblanca, Picual, Cornicabra, and Blanqueta, but much higher for Arbequina, implying that the structure of the photosynthetic apparatus is different in the latter variety. In the five varieties studied, lutein was the slowest carotenoid to be degraded, so that its percentage in the fruits increased with ripening, whereas beta-carotene was the fastest to disappear. In ripe fruits covered with anthocyanins, chloroplast pigments were retained in both skin and pulp, with the rate of disappearance being much higher in the latter.     

Variation and correlation analysis of flavonoids and carotenoids in Korean pigmented rice (Oryza sativa L.) cultivars

Flavonoids and carotenoids of pigmented rice ( Oryza sativa L.), including five black cultivars and two red cultivars, from Korea were characterized to determine the diversity among the phytochemicals and to analyze the relationships among their contents. Black cultivars were higher in flavonoids and carotenoids than the red and white cultivars. The profiles of eight phytochemicals identified from the rice grains were subjected to principal component analysis (PCA) to evaluate the differences among cultivars. PCA could fully distinguish between these cultivars. The Heugjinjubyeo (BR-1) and Heugseolbyeo (BR-2) cultivars were separated from the others based on flavonoid and carotenoid concentrations. Flavonoid contents had a positive correlation with carotenoid contents among all rice grains. The BR-1 and BR-2 cultivars appear to be good candidates for future breeding programs because they have simultaneously high flavonoid and carotenoid contents.