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.     

Determination of carotenoids in Dunaliella salina cultivated in Taiwan and antioxidant capacity of the algal carotenoid extract

A simple HPLC method with good separation efficiency was developed to determine all-trans and cis forms of carotenoids in Dunaliella salina cultivated in Taiwan. The analysis used a C30 column (250 × 4.6 mm, 5 μm) and an isocratic solvent system (flow rate = 1 mL/min) mixing methanol–acetonitrile–water (84/14/2, v/v/v) and methylene chloride, (75/25, v/v). Carotenoids were detected at 450 nm. Moreover, the antioxidant capacities of the algal carotenoid extract were also evaluated with Trolox equivalent antioxidant capacity (TEAC) assay, reducing power and 2,2-diphenyl-2-picrylhydrazyl hydrate (DPPH) radical scavenging assay. Results showed that 7 carotenoids in the algal extract could be separated simultaneously within 30 min and the total amount of them was 290.77 mg/g algae. The contents of all-trans-β-carotene and 9- or 9′-cis-β-carotene, the major carotenoids in the algae, were 138.25 and 124.65 mg/g algae, respectively. The contents of all-trans-lutein, all-trans-zeaxanthin, 13- or 13′-cis-β-carotene, all-trans-α-carotene and 9- or 9′-cis-α-carotene were 6.55, 11.27, 4.95, 2.69, and 2.41 mg/g algae, respectively. The algal carotenoid extract had significantly higher antioxidant activity than all-trans forms of α-carotene, β-carotene, lutein and zeaxanthin in all antioxidant assays. The cis forms of carotenoids, especially 9- or 9′-cis-β-carotene, might play crucial roles for the antioxidant capacities of the algal extract.     

Effect of processing and oil type on carotene bioaccessibility in traditional foods prepared with flour and puree from orange-fleshed sweetpotatoes

Consumption of Orange-Fleshed sweetpotato (OFSP) and products as source of provitamin A is being promoted to tackle vitamin A deficiency (VAD) in sub-Saharan Africa. However, limited information is available on β-carotene retention in foods and delivery after digestion. The study assessed carotene retention and bioaccessibility following in vitro digestion on traditional foods having OFSP among the ingredients. Sunflower oil, margarine and beef fat were evaluated on their effect on β-carotene bioaccessibility. Porridge and chapatis were prepared with either OFSP puree or flours in the formulations. Carotene retention was highest in chapatis (83%) as compared to porridge (65%). Micerallisation efficiency of all-trans β-carotene was comparable between similar products but greater in chapatis (62%) than porridge (11%). Sunflower oil had the highest all-trans β-carotene bioaccessibility compared to margarine and beef fat. The results support the promotion of consumption of OFSP-based products as good source of provitamin A to fight VAD.     

Genetic variability in Musa fruit provitamin A carotenoids,lutein and mineral micronutrient contents

Bananas and plantains (Musa spp.) are a staple food for millions of impoverished people and as such are an important source of vitamins and micronutrients. To evaluate the potential of Musa spp. to meet dietary micronutrients requirements, we have screened 171 different genotypes for fruit provitamin A carotenoids (pVACs) contents, and a subset of 47 genotypes for macro- and micro-mineral (iron and zinc) contents using standardised sampling and analytical protocols. The results indicate that there is substantial variability in mean fruit pulp pVACs contents between cultivars, and that cultivars with a high fruit pVACs content are widely distributed across the different genome groups but only at a low frequency. The introduction of such high pVACs cultivars has much potential for improving the vitamin A nutritional status of Musa-dependent populations at modest and realistic fruit-consumption levels. In contrast, fruit pulp mineral micronutrient contents (iron and zinc), were low and showed limited inter-cultivar variability, even for genotypes grown under widely-differing environments and soil types. Results are discussed within the framework of the development of strategies to improve the nutritional health and alleviation of micronutrient deficiencies within Musa-consuming population groups.     

Changes in β-carotene bioaccessibility and concentration during processing of carrot puree

The effect of some process and product factors (addition of olive oil, high pressure homogenisation, subsequent thermal or high pressure pasteurisation) on β-carotene bioaccessibility and isomerisation of carrot puree was investigated. High pressure homogenisation could improve β-carotene bioaccessibility by disrupting cells, but only at a pressure higher than 50 MPa. Softening of the cell walls during a subsequent thermal pasteurisation resulted in a further increase in β-carotene bioaccessibility. Unfortunately, the high temperature also induced formation of some undesirable cis-isomers. As cell walls are probably strengthened by high pressure, high pressure pasteurisation could not positively affect the amount of bioaccessible β-carotene. Moreover, the high pressure process induced β-carotene oxidation. A positive effect of olive oil on β-carotene bioaccessibility could only be noticed in combination with a thermal pasteurisation process when β-carotene was solubilised in the oil droplets. Under high pressure however, oil can be crystallised which hinders the solubilisation of β-carotene.     

Isomerisation kinetics and antioxidant activities of β-carotene in carrots undergoing different drying techniques and conditions

Carrots are known as a natural source of β-carotene. In order to preserve the latter, carrots must generally be processed, and drying is one of the most common methods for processing carrots. During drying β-carotene in carrots suffers degradation. β-Carotene degradation is generally due to thermal degradation and isomerisation. In this work, the drying kinetics as well as the isomerisation kinetics and antioxidant activities of β-carotene in carrots undergoing hot air drying, vacuum drying and low-pressure superheated steam drying (LPSSD) were determined within the temperature range of 60–80 °C and, in the case of vacuum drying and LPSSD, at a pressure of 7 kPa. A high performance liquid chromatography (HPLC) method was used to determine the β-carotene contents and its isomerisation kinetics, while the antioxidant activities of various combinations of all-trans- and cis-forms of β-carotene in carrots were evaluated using the Trolox equivalent antioxidant capacity (TEAC) assay.     

Recent advances in banana (musa spp.) biofortification to alleviate vitamin A deficiency

Abstract

Vitamin A deficiency (VAD) is one of the most prevalent micronutrient deficiencies that disproportionately affects low income populations in developing countries. Traditional breeding and modern biotechnology have significant potential to enhance micronutrient bioavailability in crops through biofortification. Bananas (Musa spp.) are economically important fruit crops grown throughout tropical and sub-tropical regions of the world where VAD is most prevalent. Some banana genotypes are rich in provitamin A carotenoids (pVACs), providing an opportunity to use bananas as a readily available vehicle for provitamin A delivery. This review summarizes the progress made in carotenoid research in bananas relative to banana diversity and the use of conventional breeding and transgenic approaches aimed at banana biofortification to address vitamin A deficiency. Existing reports on sampling strategies, pVAC retention and bioavailability are also evaluated as essential components for a successful banana biofortification effort. The wide variability of pVACs reported in banana cultivars coupled with recent advances in unraveling the diversity and genetic improvement of this globally important but often-neglected staple fruit crop underscores their importance in biofortification schemes.     

Bioavailability of β-carotene isomers from raw and cooked carrots using an in vitro digestion model coupled with a human intestinal Caco-2 cell model

Not only is there limited information in the literature regarding the β-carotene (BC) isomer profile of micelles from digested foods; few studies have looked at their subsequent uptake and transport by human intestinal Caco-2 cells. Therefore, the aims of the present study were, first, to assess the profile of BC isomers in micelles from digested raw and cooked carrots; and, second, to determine their cellular uptake and transport. Greater amounts of all-trans-, 13-cis- and 15-cis-BC isomers were present in the micelles of cooked carrots compared with raw carrots. Furthermore, micelle fractions obtained from the most highly processed (pureed) carrots had greater (P < 0.05) amounts of all-trans-, 13-cis- and 15-cis-BC compared with those derived from raw and boiled carrots. A similar trend was seen with BC isomer uptake and transport. Our data suggest that the food matrix and degree of processing play important roles on carotenoid isomerization and bioavailability.     

Freshwater fish as a dietary source of vitamin A in Cambodia

Vitamin A deficiency is a public health problem among children and women. Common Cambodian fish species were sampled and screened for vitamin A content. Contents of vitamin A-active compounds (all-trans retinol, all-trans dehydroretinol, 13-cis retinol, 13-cis dehydroretinol and β-carotene) were determined by high-performance liquid chromatography in samples of raw, whole fish from 29 fish species and in raw, edible parts from 24 species. Replicate samples were analysed in seven selected species. Two species, Parachela siamensis and Rasbora tornieri had very high vitamin A contents >1500 RAE/100 g raw, whole fish, and six species (Barbodes altus, Barbodes gonionatus, Dermogenys pusilla, Puntioplites proctozysron and Thynnichthys thynnoides) had high contents of 500–1500 RAE/100 g raw, whole fish. Two species, Puntioplites proctozysron and Thynnichthys thynnoides had high vitamin A contents in raw, edible parts, after employing traditional cleaning practices. (RAE: The amount of vitamin A active compounds in food is expressed as retinol activity equivalents (RAE), defined as the bioefficacy relative to all-trans-retinol [ West, C. E., & Eilander, A. (2002). Consequences of revised estimates of carotenoid bioefficacy for the control of vitamin A deficiency in developing countries. Journal of Nutrition, 132, 2920S–2926S]. Dehydroretinoids (vitamin A₂) are not converted to all-trans-retinol but have similar metabolic functions. In this paper, RAE refers to the functional bioefficacy as defined by Brouwer et al. [ Brouwer, I. A., Dusseldorp, M. V., West, C. E., & Steegers-Theunissen, R. P. M. (2001). Bioavailability and bioefficacy of folate and folic acid in man. Nutrition Research Review, 14, 267–293]).     

Nanoemulsion delivery systems: Influence of carrier oil on β-carotene bioaccessibility

Consumption of carotenoids may reduce the incidences of certain chronic diseases, but their use in foods is currently limited because of their poor water-solubility, low bioavailability and chemical instability. We examined the impact of carrier oil type on the bioaccessibility of β-carotene encapsulated within nanoemulsion-based delivery systems. Oil-in-water nanoemulsions (d < 200 nm) were formed using a non-ionic surfactant (Tween 20) as emulsifier and long chain triglycerides (LCT), medium chain triglycerides (MCT) or orange oil as carrier oils. The influence of carrier oil type on β-carotene bioaccessibility was established using an in vitro model to simulate the oral, gastric and small intestinal phases of the gastrointestinal tract. The rate and extent of free fatty acid production in the intestine decreased in the order LCT ≈ MCT ? orange oil; whereas β-carotene bioaccessibility decreased in the order LCT ? MCT > orange oil. The bioaccessibility of β-carotene was negligible (≈0%) in orange oil nanoemulsions because no mixed micelles were formed to solubilise β-carotene, and was relatively low (≈2%) in MCT nanoemulsions because the mixed micelles formed were too small to solubilise β-carotene. In contrast, β-carotene bioaccessibility was relatively high (≈66%) in LCT nanoemulsions. Our results have important implications for the design of effective delivery systems for encapsulation of carotenoids and other lipophilic bioactive components.     

Changes in carotenoids during processing and storage of pumpkin puree

Changes in the contents of carotenoids and their true retentions (% TR) during the production of puree of Cucurbita moschata ‘Menina Brasileira’ and of Cucurbita maxima ‘Exposição’ pumpkins and the stability of such compounds during 180 days of storage were monitored by liquid chromatography coupled with a photodiode array detector. Cooking caused higher losses than commercial sterilisation. High losses of xanthophylls such as lutein and violaxanthin were noted during processing and storage of pumpkin puree. Such losses show the low stability of these compounds. The major carotenoids, pro-vitamin A carotenes, namely, α-carotene and all-trans-β-carotene for C. moschata ‘Menina Brasileira’ and all-trans-β-carotene for C. maxima ‘Exposição’ obtained high retentions (>75%) after processing. A slight degree of isomerisation of β-carotene was noted in the puree samples, but with low concentrations of cis-isomers. Storage for 180 days did not significantly affect (P ? 0.05) the concentrations of these carotenoids.     

Vitamin A activity in foods of animal origin

The vitamin A activity of 246 samples of 59 different foods of animal origin was determined by high performance liquid chromatography. Wherever possible, the amounts of all-trans retinol, 13-cis- retinol, dehydroretinol, retinaldehyde and β-carotene were measured. The relative activity of these components is discussed, and the results compared with previous estimates of the vitamin A content of these foods.     

An in vitro model to study the intestinal absorption of carotenoids

An in vitro cell culture model to assess the intestinal absorption of carotenoids is described. When supplemented with taurocholate and oleic acid, differentiated Caco-2 cells on membranes are able to produce chylomicrons. Under conditions mimicking the in vivo postprandial state, Caco-2 cells take up carotenoids and secrete them incorporated into chylomicrons; the extents of absorption of all-trans, 9-cis, 13-cis β-carotene, α-carotene, lutein, lycopene were 11%, 2%, 3%, 10%, 7%, and 3%, respectively. Saturation of β-carotene transport through Caco-2 cells occurred at concentrations (>15 μM) higher than “physiological” concentrations. Finally, retinol supplementation resulted in an 1.7-fold increase in β-carotene transport. The data suggest that (1) the intestinal absorption of carotenoids is facilitated by the participation of a specific epithelial transporter and (2) retinol promotes β-carotene incorporation into larger, retinyl ester-enriched chylomicrons. The present in vitro cell culture system is a relevant model to study the intestinal absorption of carotenoids at the molecular level.     

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.     

Degradation compounds of carotenoids formed during heating of a simulated cashew apple juice

The influence of organic acid and heating treatments on carotenoid degradation on a simulated cashew apple juice was assessed by high performance liquid chromatography coupled with a photodiode array and mass spectrometry detectors. A total of nineteen carotenoids were separated in unheated simulated cashew apple juice, with all-trans-β-cryptoxanthin and all-trans-β-carotene as the major ones. As a consequence of heating, five xanthophylls disappeared, whereas two new cis isomers and five epoxide or furanoid-derivatives were formed and the levels of all cis isomers increased. In addition, 12′-apo-β-carotenal was formed at 90 °C. Two oxidation compounds (12′-apo-β-carotenal and 5,6-epoxy-β-cryptoxanthin) were formed after β-cryptoxanthin heating at 90 °C in an aqueous-based system. In all systems, the amounts of total carotenoids lost were not compensated by those formed. These facts indicated that isomerisation and oxidation to both coloured and non-coloured compounds were the main reactions occurring during heating of carotenoids in aqueous-based and juice systems.     

Thermal degradation of all-trans-β-carotene in the presence of phenylalanine

Solutions of all-trans-β-carotene in liquid paraffin were stirred under air at 210°C for 15 min in the presence and absence of various amino acids. After cooling, the absorbance values at 460 nm of model systems containing all-trans-β-carotene and phenylalanine, cysteine or tryptophan were 34–58 times higher than that obtained on heating in the absence of an amino acid. Based on absorbance values at 420 nm, 86% of the coloured material present on heating the all-trans-β-carotene-phenylalanine model system at 210°C for 15 min was retained by the liquid paraffin on methanol extraction. The liquid paraffin phase remaining after methanol extraction was analysed by reverse-phase HPLC and three major components were identified as all-trans-β-carotene and two degradation products of all-trans-β-carotene, ie 9-cis-β-carotene and a furanoid diepoxy derivative. None could be detected on heating all-trans-β-carotene alone, suggesting that its degradation was retarded on heating in the presence of phenylalanine.     

Beta-carotene isomerisation in mango puree as influenced by thermal processing and high-pressure homogenisation

The present study describes a case study on mango puree, in which focus is given to the effect of thermal processing (100–130 °C, 0–80 min) and high-pressure homogenisation (0–1300 bar) on the isomerisation of β-carotene. Both unit operations are of relevance for the production of mango puree. β-Carotene is an essential micronutrient which is present in a high amount in most mango cultivars, and it is important for human health due to its antioxidant and provitamin A capacity. It is known that these health-related properties of β-carotene are negatively affected by the conversion to cis-isomers. The results have shown that during high-pressure homogenisation of mango puree, β-carotene isomerisation was negligible. During thermal processing, on the other hand, an increase in β-carotene cis-isomer formation with increasing treatment intensity could be observed, although high temperatures and/or long treatment times were required to observe clear additional isomer formation. From a kinetic point of view, a fractional conversion model could be used to model the all-trans-β-carotene isomerisation in mango puree in the temperature and time range studied. In general, it can be concluded that a high percentage of β-carotene is present as cis-isomers in raw mango puree. Furthermore, only intense thermal processing of mango puree leads to the formation of additional cis-isomers in relevant amounts.     

Composition of Carotenoids in <Emphasis Type="Italic">Scenedesmus protuberans</Emphasis>: Application of Chromatographic and Spectroscopic Methods

This study aimed to identify and determine the carotenoids from green microalga, Scenedesmus protuberans using analytical techniques. Identification of carotenoids was realized by comparing their absorption and mass spectral data with those of reference standards available and reported values. Chromatographic data were then combined with the spectroscopic information. The separation of carotenoids was achieved by C₃₀ column and high-performance liquid chromatography-diode array detection was used for their determination. In the present work, the carotenoid content of S. protuberans was found to be 1.45 mg/g of violaxanthin, 2.47 mg/g of all-trans-lutein, 0.15 mg/g of all-trans-α-carotene, 0.55 mg/g of all-trans-β-carotene, and 0.20 mg/g of 9 or 9′-cis-β-carotene. Due to lack of their standards, the amount of all-trans-loroxanthin and cis-isomers of other carotenoids could not be quantified. In order to validate the method, Certified Reference Material (BCR 485-Mixed vegetables) was used. In conclusion, this study can serve as a reference for the analysis of carotenoids in other microalgae.     

Effects of Chinese domestic cooking methods on the carotenoid composition of vegetables in Taiwan

Carotenoids, lutein, zeaxanthin, and β-carotene, common in many vegetables, have been shown to be important in visual health-promoting. However, cooking practice has a great impact on these bioactive compounds. The objective of this work was to evaluate the effect of three common Chinese domestic cooking methods including boiling, stir-frying, and deep-frying on the composition of lutein, zeaxanthin, β-carotene and their trans/cis (E/Z) isomers of vegetables. Among the 25 fresh vegetables commonly consumed in Taiwan, it was found that cilantro, Thai basil leaves, sweet potato leaves, and choy sum contained the 1st–4th highest amount of total carotenoids content (TCC), respectively. Boiling preserved the majority of carotenoids, whereas stir-frying and deep-frying significantly decreased TCC and (all-E)-forms of carotenoids. In addition, (all-E) – and (13Z)-/(13′Z)-carotenoids exhibited similar trends during boiling, since the boiling time required to reach maximum concentration for (all-E)-carotenoids was almost the same as that for (13Z) – plus (13′Z)-isomers. An overall increase of (9Z)/(9′Z)-carotenoid isomers was observed in all boiled vegetables, probably because of E/Z isomerization, which could partially result in the formation of more bioactive compounds such as (9Z)-β-carotene. Boiling may be the preferred practice of domestic cooking methods to preserve carotenoids in vegetables.     

Determination of thermal oxidation and oxidation products of β-carotene in corn oil triacylglycerols

A reversed phase HPLC-DAD coupled to APCI-MS method is reported for the determination of the degradation of β-carotene and characterization of the oxidation products in corn oil. Corn oil containing β-carotene was oxidized in the Rancimat at 110 °C from 1 to 14 h. A significant degradation of β-carotene was observed in corn oil during accelerated thermal oxidation. A total of eight different oxidized products of β-carotene were identified, which includes 8′-apo-β-carotenal, 6′-apo-β-carotenal, 5,6-epoxy-8′-apo-β-carotenal, β-carotene-2,2′-dione, 13-Z-5,6,5′,6′-diepoxy-β-carotene, all-E-5,8-epoxy-β-carotene, all-E-5,6-epoxy-β-carotene, and 15-Z-5,6-epoxy-β-carotene. Corn oil triacylglycerols (TAGs) oxidation products were also identified using isocratic HPLC–ESI-MS. It was found that β-carotene promoted oxidation of TAGs especially at longer exposure times. For the first time two new classes of oxidized TAGs have been identified in corn oil, which were epidioxy bis-hydroperoxides and hydroxy bis-hydroperoxides. Other oxidation products of TAGs were mono-epoxides, mono-hydroperoxides, and epoxy hydroperoxides. It was found that complementing the HPLC-DAD method for β-carotene with ESI-MS method for TAGs oxidation provides a comprehensive set of analytical tools to characterize carotenoids and triacylglycerols oxidation and degradation in edible oils.