Effects of supercritical CO2 fluid parameters on chemical composition and yield of carotenoids extracted from pumpkin

Pumpkin is a traditional food that is grown extensively worldwide and is believed to be beneficial to human health due to its high contents of carotenoids. The carotenoids in pumpkin were extracted by organic solvents and by supercritical carbon dioxide (SC-CO₂), and then they were identified, quantified, and compared. β-carotene (31 to 40 g per 100 g of total carotenoids) was the predominate carotenoid in pumpkin. Lutein and lycopene contents were much higher in SC-CO₂ extracts than those in organic solvent extract. Cis-β-carotene increased by more than two times in the SC-CO₂ extracts, even at a relatively low temperature of 40 °C, over those in the solvent extracts, indicating both enhanced solubility and isomerization from trans- to cis-β-carotene. The influences of modifier (10 mL/100 mL), temperature (40-70 °C), and pressure (25-35 MPa) of SC-CO₂ extraction on the change of carotenoid yields were also investigated. The highest yield (109.6 μg/g) was obtained at 70 °C and 35 MPa, with a 73.7% recovery. Selective extraction could be achieved by adjusting the temperature and pressure. Higher proportions of all-trans-β-carotene extracts were achieved at 40 °C under both 25 MPa and 35 MPa conditions. In order to extract more cis-isomers, a higher temperature of 70 °C was preferred.     

Degradation of carotenoids in pumpkin (Cucurbita maxima L.) slices as influenced by microwave vacuum drying

The aim of this study was to investigate the influence of microwave vacuum drying on carotenoids in pumpkin (Cucurbita maxima L.) slices. Carotenoids were measured using the reverse-phase high-performance liquid chromatography technique. It was shown that compared with hot air drying, microwave vacuum drying inhibited color changes and significantly (p < 0.05) improved total carotenoid retention (89.1%) in pumpkin slices. During the microwave vacuum drying process, microwave power had an important effect on total carotenoid and all-trans carotenoids. As microwave power increased, the total carotenoid content significantly decreased (p < 0.05), and the levels of individual carotenoids, including all-trans-α-carotene, all-trans-β-carotene, and all-trans-lutein, generally decreased. However, there was an overall upward trend for the levels of 13-cis-β-carotene, 15-cis-β-carotene, 9-cis-β-carotene, and 9-cis-α-carotene. The trans carotenoid quality of the finished products was improved within a certain range of vacuum levels. In addition to the degradation induced by microwave energy, isomerization was considered to be responsible for the loss of all-trans carotenoids. These results indicated that inappropriate drying methods and conditions might result in high losses of all-trans carotenoids in pumpkins.     

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 and isomerization of chlorophyll a and β-carotene as affected by various heating and illumination treatments

The degradation and isomerization of β-carotene and chlorophyll a as affected by oven-heating, reflux-heating, iodine-catalysed illumination, and non-iodine-catalysed illumination, were studied. Results showed that the degradations of both total β-carotene and chlorophyll a may fit the first-order model under either heating or illumination treatment. 13-cis-β-Carotene and 13,15-di-cis-β-carotene were the major cis isomers of β-carotene formed during oven heating, while 13-cis-β-carotene was favoured during reflux heating. For illumination with or without iodine as catalyst, 13,15-di-cis-β-carotene was the major cis isomer of β-carotene formed. The formation of 13,15-di-cis-β-carotene may be due to conversion of either 13-cis- or 15-cis-β-carotene. No epimerization of chlorophyll a was observed as a result of illumination.     

Carotenoid Photostability in Raw Spinach and Carrots During Cold Storage

A reverse-phase gradient HPLC method was developed for separating xanthophylls, chlorophylls, carotenes and cisβ-carotene isomers from raw spinach and carrots. The effect of dark and lighted cold storage on pigment stability was studied. Light promoted pigment losses in raw spinach. Degradative losses at 8 days ranged from 60% for violaxanthin to 22% for lutein. Dark, cold storage did not affect carotenoid levels except for all-transβ-carotene which showed an 18% loss at 8 days. In raw carrots, neither lighted nor dark cold storage affected major carotenoids. In spinach, the isomeric distribution of β-carotene showed strong linear correlations between trans and cis forms.     

Effect of processing variables on the stability of β-Carotenes and xanthophylls of dehydrated parsley

Curled- and plain-leaf parsley were analysed for β-carotene, xanthophyll and the stereo-isomers of β-carotene, before and after various combinations of freezing, blanching, sulphiting and air-drying. Heating of parsley by blanching or by dehydration of unblanched samples generally resulted in an increased retention of β-carotene. Dried samples contained a higher percentage of the cis-isomer and neo-β-carotene b but a lower percentage of the all-trans-isomer. Freezing unblanched parsley reduced total carotene content 8–13 %. Xanthophyll changes were similar to those for total carotene except that dried samples contained 2–28 % less xanthophyll than fresh parsley. Sulphited samples retained 8–30 % more carotenoid pigments than the non-sulphited samples. Total β-carotene of dried parsley stored for 3 months in air and in nitrogen at 22° decreased 10-15%and 3·5-10%, respectively, while that stored at –18° showed no change. The stereo-isomer ratio did not change. Blanching had no apparent effect on carotene stability in storage. Carotene loss in fresh parsley, additional extractions, and enzymic destruction of added carotene were also investigated.     

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.     

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.     

Carotenoid Composition and Vitamin A Value of Fresh and Pasteurized Cashew-Apple (Anacardium occidentale L.) Juice

The carotenoid composition of the red and yellow cashew-apples and two commercial brands of pasteurized juice were determined by column and thin-layer chromatography, visible absorption spectroscopy and specific chemical reactions. Alpha-carotene, β-carotene, ζ-carotene, cis-β-carotene, cryptoxanthin, aurochrome, cryptochrome, and auroxanthin were identified in all samples, with β-carotene as the predominant pigment. Color differences reflected the total amount, not the type of carotenoid present. Geographic effects on the pigment composition did not appear to be appreciable. In the processed juice, the total carotenoid content and vitamin A value were lower but the epoxide level higher. No quantitative differences were noted among lots of the same brand or between brands of pasteurized juice.     

Use of Multi-response Modelling to Investigate Mechanisms of β-Carotene Degradation in Dried Orange-Fleshed Sweet Potato During Storage: from Carotenoids to Aroma Compounds

In order to give insight into β-carotene degradation mechanism during the storage of dried orange-fleshed sweet potato, and particularly into the role of isomers and norisoprenoids formation, multi-response kinetic modelling was applied. Determination of degradation compounds were carried out by HPLD-DAD and SPME-GC-MS as a function of time between 10 and 40 °C and at four water activities from 0.13 to 0.76. Kinetic modelling was developed assuming first-order reactions and by using mass balance. Eight compounds, namely, two isomers (9-cis- and 13-cis-β-carotene), two β-carotene epoxides (β-carotene 5,6 and 5,8 epoxide) and four volatile compounds (β-cyclocitral, β-ionone, 5,6-epoxy-β-ionone and dihydroactinidiolide), were integrated into two theoretical reaction schemes. The different models were discriminated according to goodness of fit to experimental data. This work showed that: (1) the formation of cis-isomers from β-carotene preceded oxidation, (2) β-cyclocitral arose directly from β-carotene scission while the other norisoprenoids resulted from β-carotene epoxide degradation, (3) cis-isomers were high reactive compounds. Temperature had a major influence on reaction rates k while water activities only impacted k at values under 0.51. Therefore, multi-response modelling is not only a tool to predict β-carotene degradation but a interesting way to select the appropriate degradation scheme based on the different options presented in literature.     

Apparent solubility of lycopene and β-carotene in supercritical CO2, CO2 + ethanol and CO2 + canola oil using dynamic extraction of tomatoes

Lycopene and β-carotene were extracted from freeze-dried tomatoes (skin + pulp) with pure SC CO₂ and SC CO₂ + 5% w/w co-solvent at 40 °C, 400 bar and flow rates of 0.5 and 1.2 L/min. The apparent solubility of lycopene and β-carotene in the multicomponent complex system was determined from dynamic extraction experiments using a laboratory-scale supercritical extraction system. Solubility of pure lycopene and β-carotene in SC CO₂ (binary system) was reported in the literature to be of the order of 10⁻⁶ mole fraction. The apparent solubility of lycopene extracted from tomatoes with SC CO₂ (multicomponent complex system) under the same conditions was almost one order of magnitude smaller. The apparent solubility obtained using oil as a co-solvent was higher than that obtained with ethanol as a co-solvent or pure SC CO₂. The differences in solubility are mainly due to the polarity of the co-solvent and the impact of the tomato matrix in the multicomponent complex system.     

Antioxidant activity of tomato lipophilic extracts and interactions between carotenoids and α-tocopherol in synthetic mixtures

In the present study we assayed the antioxidant activity of lipophilic extracts obtained from different tomato varieties. The results showed that cherry tomatoes, characterized by a high carotenoid content, had the highest antioxidant activity. A quantitative analysis of lycopene, β-carotene, lutein and α-tocopherol was also performed and the correlation between the antioxidant content and the antioxidant activity was estimated. The highest correlation coefficient was found for lycopene (R² = 0.9236, P ≤ 0.001). The analysis of two-component mixtures containing α-tocopherol and carotenoids showed that significant synergism occurred for all the combinations which contained α-tocopherol and β-carotene mixed together. The highest synergistic effects were detected for α-tocopherol-lycopene mixtures, which were the most efficient combinations tested in the present study. The analysis of the carotenoid combinations indicated that synergism occurred for lycopene-β-carotene, lycopene-lutein and lutein-β-carotene mixtures. The analysis of four-component mixtures did not show statistically significant synergistic effects.     

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.     

Production of a carotenoid-rich product by alginate entrapment and fluid-bed drying of Dunaliella salina

A new carotenoid-rich product was formed by entrapment of Dunaliella salina in calcium alginate beads of different alginate concentrations, followed by drying in a fluid-bed dryer. The drying process yielded β-carotene recoveries of between 79 and 89% and produced a change in the 9-cis/all-trans ratio of β-carotene isomers. The carotenoid stability of the product was dependant on both the storage conditions and alginate content (range 3·3–7·3 g litre⁻¹) of the beads. In the presence of light and oxygen total carotenoid degraded following a first order kinetic model with degradation constants between 0·016 and 0·039 days⁻¹, with the lowest degradation occurring with the lowest alginate concentration. Product stored in the dark and flushed with nitrogen produced first-order degradation constants of 0·012 and 0·020 days⁻¹ for the two higher alginate concentrations; that with the lowest alginate content showed no noticeable degradation after 58 days storage. During storage, the 9-cis isomer was significantly more unstable showing a relative loss under all conditions, degrading almost completely when stored in the presence of light and oxygen and reaching an equilibrium ratio with the all-trans isomer when stored in the dark and flushed with nitrogen. © 1998 SCI.     

Characterisation of oleoresins extracted from tomato waste by liquid and supercritical carbon dioxide

Tomato waste is characterised by high amounts of bioactive compounds, such as carotenoids, polyphenols and essential fatty acids. In this work, oleoresins were produced from tomato waste (skins + seeds) and skins using carbon dioxide under liquid and supercritical conditions. The use of ethanol as a co-solvent was also evaluated. Compositions of the oleoresins were analysed and compared to oleoresin obtained by means of an organic solvent, which was used as the control. Oleoresin extraction from the waste by liquid and supercritical CO₂ provided the highest yields (12.5%–12.9%). The extracts obtained from the skins with supercritical CO₂ without ethanol had the highest content of α- and γ-tocopherols (80 and 575 mg/100 g oleoresin, respectively), while lycopene and β-carotene were found at 205 and 75 mg/100 g oleoresin, respectively. The highest quantity of polyphenols (9305 mg GAE/100 g oleoresin) was found in the oleoresin extracted from skin by supercritical CO₂ with ethanol. All oleoresins were characterised by a high antioxidant activity, which improved with the synergy of carotenoids, tocopherols and polyphenols.     

Suppressive effect of carotenoid extract of Dunaliella salina alga on production of LPS-stimulated pro-inflammatory mediators in RAW264.7 cells via NF-κB and JNK inactivation

The carotenoid extract from Dunaliella salina was used to evaluate the suppressive effects on lipopolysaccharide (LPS)-induced pro-inflammatory mediators in RAW264.7 cells. The extract composed all-trans forms of α-carotene (28.8 mg/g extract), β-carotene (471.1 mg/g extract), lutein (7.1 mg/g extract) and zeaxanthin (7.2 mg/g extract), 13- or 13′-cis-β-carotene (12.1 mg/g extract), 9- or 9′-cis-α-carotene (19.1 mg/g extract) and 9- or 9′-cis-β-carotene (440.3 mg/g extract) dose-dependently reduced the production of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α), the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and the secretion of nitric oxide (NO) and prostaglandin E₂ (PGE₂) in LPS-activated RAW264.7 cells. Its attenuation of LPS-induced inflammatory responses was closely related to inhibition of the nuclear NF-κB p50 subunit translocation by blocking inhibitor of κBα (IκB) phosphorylation and degradation correlated with suppressing IκB kinase (IKK) α/β phosphorylation, as well as down-regulation of the c-Jun NH₂-terminal kinase (JNK) activation.     

Antioxidant activity of extracts obtained by different isolation procedures from some aromatic herbs grown in Lithuania

An increasing demand for natural additives has shifted the attention from synthetic to natural antioxidants. The present work examines the potential of some aromatic herbs grown in Lithuania as a source of natural antioxidants: marjoram (Majorana hortensis Moench), catnip (Nepeta cataria L), oregano (Origanum vulgare L), lavender (Lavandula angustifolia Mill), thyme (Thymus vulgaris L), hyssop (Hyssopus officinalis L), anise hyssop (Lophantus anisatus Benth), and sage (Salvia officinalis L). Dried herb samples were submitted to extraction with supercritical CO₂, acetone or methanol/water and hydrodistillation. Deodorised herb samples (after removal of essential oil) were also extracted with acetone. The antioxidant activity of these extracts, essential oils and dried deodorised aqueous extracts was assessed by the β-carotene bleaching test (diffusion and spectrophotometric methods). The highest yields of extracts were obtained using polar solvents. Thyme and sage acetone oleoresins showed high antioxidant activity in the tests performed and were regarded as the most promising sources. © 1998 SCI.     

Effect of Natural Extracted Antioxidants from Eriobotrya japonica (Lindl.) Fruit Skin on Thermo Oxidative Stability of Soybean Oil During Deep Frying

The effects of ultrasound-assisted, supercritical CO₂ and solvent extraction techniques on antioxidant activity of Eriobotrya japonica (Lindl.) skin extracts were investigated using 2, 2-diphenyl-1-picrylhydrazyl, β-carotene, and Rancimat assays. Solvent extract of skin at 400 ppm had the highest antioxidant activity. Solvent extraction was the most effective method on extraction of phenols and tocopherols. Protective effects of extracts in stabilization of soybean oil during frying were determined by measuring its peroxide value, total polar compounds, carbonyl value, free fatty acids, conjugated dienes, and trienes. Results indicated that solvent extract of skin at 400 ppm exhibited stronger antioxidant activity in oil than tertiary butylhydroquinone.     

Determination of β-Carotene Stereoisomers by Spectrophotometric Analysis

A method for determining the amount of β-carotene stereoisomers using spectrophotometric analysis was developed. Water-soluble β-carotene was dissolved in pH 6.0 buffer and heated. The stereoisomers formed were separated by column chromatography and quantified using spectrophotometric analysis. The %D_B/D₁₁ value (absorbance of the cis-peak divided by the absorbance at the maximum wavelength) determined for the β-carotene solutions was found to be linearly correlated to the isomer composition (r = 0.971).     

Utilization of Jujube Fruit (Ziziphus mauritiana Lam.) Extracts as Natural Antioxidants in Stability of Frying Oil

The effects of ultrasound-assisted, supercritical CO₂ and solvent extraction methods on antioxidant activity of Ziziphus mauritiana (Lam.) extracts were investigated using 2,2-diphenyl-1-picrylhydrazyl and β-carotene assays. Ethanol-water extract of the jujube that was obtained with ultrasound-assisted extraction had the highest antioxidant activity. Ultrasound-assisted extraction was the most effective method on extraction of phenolic compounds. The effect of ultrasound extract in stability of soybean oil was compared to synthetic antioxidants by measuring total polar compounds, carbonyl value, peroxide value, free fatty acids, oxidative stability index, and conjugated dienes and trienes values. Results showed that ultrasound ethanol-water extract at 600 ppm had a higher stabilization efficiency than commercial antioxidants butylated hydroxyanisole, butylated hydroxytoluene, and tertiary butylhydroquinone.