Sources of errors in the quantitative analysis of food carotenoids by HPLC

Several factors render carotenoid determination inherently difficult. Thus, in spite of advances in analytical instrumentation, discrepancies in quantitative results on carotenoids can be encountered in the international literature. A good part of the errors comes from the pre-chromatographic steps such as sampling scheme that does not yield samples representative of the food lots under investigation; sample preparation which does not maintain representativity and guarantee homogeneity of the analytical sample; incomplete extraction; physical losses of carotenoids during the various steps, especially during partition or washing and by adsorption to glass walls of containers; isomerization and oxidation of carotenoids during analysis. on the otherhand, although currently considered the method of choice for carotenoids, high performance liquid chromatography (HPLC) is subject to various sources of errors, such as: incompatibility of the injection solvent and the mobile phase, resulting in distorted or split peaks; erroneous identification; unavailability, impurity and instability of carotenoid standards; quantification of highly overlapping peaks; low recovery from the HPLC column; errors in the preparation of standard solutions and in the calibration procedure; calculation errors. Illustrations of the possible errors in the quantification of carotenoids by HPLC are presented.

Total and individual carotenoid profiles in Solanum phureja of cultivated potatoes: I. Concentrations and relationships as determined by spectrophotometry and HPLC

Total and individual carotenoid concentrations were determined by spectro photometry and HPLC, in raw tubers of a sample of 23 accessions of Solanum phureja potatoes taken at random from the world germplasm collection following its stratification on tuber flesh color. Lutein, zeaxanthin, violaxanthin, antheraxanthin and beta-carotene were detected in all accessions and three distinct patterns of carotenoid accumulation were evidenced by cluster analysis. Accessions in group 1 showed the highest concentrations of total carotenoids (1258-1840 mu g 100 g(-1) FW) comprised largely of zeaxanthin (658-1290 mu g 100 g(-1) FW) with very low or no presence of beta-carotene (below 5.4 mu g 100 g(-1) FW). Accessions in group 2 presented moderate total carotenoid concentrations with violaxanthin, antheraxanthin, lutein and zeaxanthin as the major carotenoids. Accessions in group 3 showed low concentrations of total carotenoids (97-262 mu g 100 g(-1) FW) and very low or no zeaxanthin, with lutein and violaxanthin as the predominant carotenoids and relatively high concentrations of beta-carotene(up to 27 mu g 100 g(-1) FW). Five accessions with significant concentrations of zeaxanthin were identified with the accession 703566 showing the highest concentration (1290 p g 100 g(-1) FW). This value is to our knowledge higher than any value previously reported for potatoes, including those achieved through genetic modification. For the 23 S. phureja accessions, total carotenoid concentration was positively and significantly correlated with antheraxanthin and zeaxanthin concentrations, and negatively and significantly correlated with beta-carotene concentration. (C) 2008 Elsevier B.V. All rights reserved.

beta-carotene and other carotenoids as antioxidants

Carotenoids are natural pigments which are synthesized by plants and are responsible for the bright colors of various fruits and vegetables. There are several dozen carotenoids in the foods that we eat, and most of these carotenoids have antioxidant activity. beta-carotene has been best studied since, in most countries it is the most common carotenoid in fruits and vegetables. However, in the U.S., lycopene from tomatoes now is consumed in approximately the same amount as beta-carotene. Antioxidants (including carotenoids) have been studied for their ability to prevent chronic disease, beta-carotene and others carotenoids have antioxidant properties in vitro and in animal models. Mixtures of carotenoids or associations with others antioxidants (e.g. vitamin E) can increase their activity against free radicals. The use of animals models for studying carotenoids is limited since most of the animals do not absorb or metabolize carotenoids similarly to humans.Epidemiologic studies have shown an inverse relationship between presence of various cancers and dietary carotenoids or blood carotenoid levels. However, three out of four intervention trials using high dose beta-carotene supplements did not show protective effects against cancer or cardiovascular disease. Rather, the high risk population (smokers and asbestos workers) in these intervention trials showed an increase in cancer and angina cases. It appears that carotenoids (including beta-carotene) can promote health when taken at dietary levels, but may have adverse effects when taken in high dose by subjects who smoke or who have been exposed to asbestos. It will be the task of ongoing and future studies to define the populations that can benefit from carotenoids and to define the proper doses, lengths of treatment, and whether mixtures, lather than single carotenoids (e.g. beta-carotene) are more advantageous.

Postprandial plasma carotenoid responses following consumption of strawberries, red wine, vitamin C or spinach by elderly women

This study investigated the postprandial plasma responses of carotenoids for 24 h after feeding five specific breakfast beverages; four of which had low or no carotenoid content. In seven fasting healthy elderly female subjects a blood sample (baseline) was obtained, after which they were given a breakfast beverage, containing one of the following: 1) strawberries (240 g); 2) ascorbic acid (1250 mg); 3) spinach (294 g); 4) red wine (300 mL); and 5) control (breakfast beverage only). Blood samples were collected at 0.5, 1, 4, 7, 11, 15 and 24 h. Plasma carotenoids were measured using HPLC. No significant differences were found in the levels of the plasma carotenoids measured among the various treatments at baseline. In the spinach treatment, plasma lutein, zeaxanthin and β-carotene levels at 7, 11, 15 and 24 h were significantly higher than those at baseline, as expected. All of the carotenoids measured in the control and vitamin C treatments, at subsequent sampling times were not significantly different from those at baseline. However, for most carotenoids, strawberry and red wine feeding resulted in significantly lower carotenoids values from baseline at 11 and 15 h. Subjects who received a diet with low levels of carotenoids, but whose postprandial plasma levels of carotenoids remain steady, might be explained by a mechanism that promotes secretion of carotenoids into the circulation. Assuming that plasma carotenoids are being used over time, we hypothesize that strawberries and red wine contain some substances that interfere with the secretion of carotenoids into the circulation.

β-carotene and other carotenoids as antioxidants

Carotenoids are natural pigments which are synthesized by plants and are responsible for the bright colors of various fruits and vegetables. There are several dozen carotenoids in the foods that we eat, and most of these carotenoids have antioxidant activity. β-carotene has been best studied since, in most countries it is the most common carotenoid in fruits and vegetables. However, in the U.S., lycopene from tomatoes now is consumed in approximately the same amount as β-carotene. Antioxidants (including carotenoids) have been studied for their ability to prevent chronic disease. β-carotene and others carotenoids have antioxidant properties in vitro and in animal models. Mixtures of carotenoids or associations with others antioxidants (e.g. vitamin E) can increase their activity against free radicals. The use of animals models for studying carotenoids is limited since most of the animals do not absorb or metabolize carotenoids similarly to humans. Epidemiologic studies have shown an inverse relationship between presence of various cancers and dietary carotenoids or blood carotenoid levels. However, three out of four intervention trials using high dose β- carotene supplements did not show protective effects against cancer or cardiovascular disease. Rather, the high risk population (smokers and asbestos workers) in these intervention trials showed an increase in cancer and angina cases. It appears that carotenoids (including β-carotene) can promote health when taken at dietary levels, but may have adverse effects when taken in high dose by subjects who smoke or who have been exposed to asbestos. It will be the task of ongoing and future studies to define the populations that can benefit from carotenoids and to define the proper doses, lengths of treatment, and whether mixtures, rather than single carotenoids (e.g. β-carotene) are more advantageous.

Endogenous carotenoid concentrations in cancerous and non-cancerous tissues of gastric cancer patients in Korea

Carotenoid concentrations were measured in serum and in both non-cancerous and cancerous gastric mucosal tissues of Korean patients with gastric cancer (n = 18). Carotenoids in serum and gastric tissue were extracted with chloroform/methanol (2:1), and measured using reverse-phase high-performance liquid chromatography with a C30 column. Cryptoxanthin and β-carotene were the major carotenoids in the Korean blood and they had a median ratio of non-cancerous tissue/serum levels which was less than 1.0. No significant differences of Cryptoxanthin and β-carotene levels were found between non-cancerous and cancerous tissues. After incubation of β-carotene with gastric tissue, significantly higher levels of β-carotene breakdown products were produced in the homogenates of cancerous tissue when compared with non-cancerous tissue. Lutein, zeaxanthin and α-carotene were the minor carotenoid constituents in the blood and their median ratio of non-cancerous tissue/serum levels was greater than 1.0. Cancerous tissue had significantly lower levels of lutein, zeaxanthin and α-carotene than did non-cancerous tissue. It appears that the increased breakdown of β-carotene and cryptoxanthin in cancerous tissue can be compensated for by an increased uptake of circulating carotenoids by cancerous tissue, whereas lutein, zeaxanthin and α-carotene levels in cancerous tissue are not able to be maintained.

Carotenoid composition of hydroponic leafy vegetables

Because hydroponic production of vegetables is becoming more common, the carotenoid composition of hydroponic leafy vegetables commercialized in Campinas, Brazil, was determined. All samples were collected and analyzed in winter. Lactucaxanthin was quantified for the first time and was found to have concentrations similar to that of neoxanthin in the four types of lettuce analyzed. Lutein predominated in cress, chicory, and roquette (75.4 ± 10.2, 57.0 ± 10.3, and 52.2 ± 12.6 μg/g, respectively). In the lactucaxanthin-containing lettuces, β-carotene and lutein were the principal carotenoids (ranging from 9.9 ± 1.5 to 24.6 ± 3.1 μg/g and from 10.2 ± 1.0 to 22.9 ± 2.6 μg/g, respectively). Comparison of hydroponic and field-produced curly lettuce, taken from neighboring farms, showed that the hydroponic lettuce had significantly lower lutein, β-carotene, violaxanthin, and neoxanthin contents than the conventionally produced lettuce. Because the hydroponic farm had a polyethylene covering, less exposure to sunlight and lower temperatures may have decreased carotenogenesis.

Carotenoid composition of Brazilian fruits and vegetables

Brazil has a wide diversity of food sources of carotenoids. The updated Brazilian database consists of more than 270 items of fruits, vegetables and their prepared and processed products. The database demonstrates variations due to variety, maturity, production technique, climate and processing. Many of these foods are not found in the US and European databases. Good to rich sources (>20 μg/g) of β-carotene are: acerola, bocaiúva, mango 'Extreme' and tucumã. Sources of both α-carotene and β-carotene are buriti, carrot, Cucurbita moschata 'Menina Brasileira', 'Baianinha' and 'Goianinha', and red palm oil. Commercially produced and uncultivated or semi-cultivated leafy vegetables, C. maxima 'Jerimum Caboclo' and the hybrid Tetsukabuto, cooked broccoli are sources of lutein and β-carotene. The edible Tropaeolum majus flower is especially rich in lutein. Although many fruits have β-cryptoxanthin as principal carotenoid (e.g. caja, nectarine, peach, orange-fleshed papaya, tree tomato), the levels are below 20 μg/g. Good to rich sources of lycopene are guava and guava products, papaya, pitanga and pitanga juice, tomato and tomato products, and watermelon. Sources of zeaxanthin are rare; although the principal carotenoid of piqui, the amount is low, lower than that found in buriti.

Edible coatings from native and modified starches retain carotenoids in pumpkin during drying

The goal of this study was to evaluate the effect of edible coating pre-treatments on the retention of provitamin A during pumpkin drying. The coatings used were based on native and modified maize and cassava starch. To evaluate the effects of these coatings, slices of 'Dry Rajada' pumpkin were dried at 70 °C both with and without starch coatings applied at 30 and 80-90 °C. Carotenoid content was determined through HPLC using a C 30 column. Significant losses (12-15%) of trans-α-carotene and trans-β-carotene were observed when slices were dried without the coating. Significant improvement of carotenoid content was observed for dehydrated slices that were previously coated with a native maize starch solution at 90 °C, as well as with a modified maize starch solution at 30 °C and also with a modified cassava starch solution at 90 °C. The application of these starch solutions probably produced a more uniform film that adhered to the slices, minimizing carotenoid degradation during pumpkin drying and, as a consequence, resulting in a product that can be considered a good source of provitamin A. © 2012 Elsevier Ltd.

Antioxidant activity and determination of total phenol contents, carotenoids, beta-carotene, lycopene and zinc in white, yellow and pinkish varieties of manihot esculenta crantz

There are cassava varieties that present compounds as carotenoids, beta-carotene, lycopene and minerals important for human and animal health. The present study evaluated the antioxidant activity of the white, yellow and pinkish varieties of Manihot esculenta, by mean of the DPPH test and by the ferrous ion-chelating activity. Furthermore, the total phenols, carotenoids, beta-carotene, lycopene and zinc contents were also determined. Utilizing the DPPH test it was possible to find that extracts of boiled samples presented higher antioxidant activity (89.53% -pinkish) in comparison to the fresh samples (1.97% -white). For the ferrous ion-chelating test, the highest activity was found for the boiled pinkish variety extract (63.43%) and the lowest was for fresh yellow extract (17.34%) the white sample did not present activity. The highest concentration of total phenols and zinc content was obtained for the boiled pinkish variety extract 136.12 mg EAG/g of extract and 0,811ppm, respectively, in the concentration of 1000 mu g/mL. The pinkish variety presented also higher quantity of pigments, including carotenoid (29.40 mu g/g), beta-carotene (9.14 mu g/100g) and lycopene (68.92%). According to the results obtained in this study it was possible to conclude that the yellow and pinkish varieties of M. esculenta present quantity of phenolic compounds and minerals sufficient to attribute the antioxidant activity and may thus contribute to reduce oxidative damage and be used as nutraceuticals or directly ingested in the diet to maintain good health.

Novel bioassay for the discovery of inhibitors of the 2-C-Methyl-D-erythritol 4-Phosphate (MEP) and terpenoid pathways leading to carotenoid biosynthesis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Phenolic compounds, carotenoids, tocopherols and fatty acids present in oils extracted from palm fruits

This review aimed to discuss the main bioactive compounds present in oils extracted from palm trees, indicating possible applications for the same. The bioactive compounds approached were: phenolic compounds, carotenoids, tocopherols, mono and polyunsaturated fatty acids. It is growing the search for food, especially from plant origin, characterized by the presence of such substances due to their benefi ts. Many studies show palm species as important sources of bioactive compounds and essential fatty acids. Therefore it is important to study the fruit characterization and composition of the oil from palm species of different regions that are still poorly investigated, seeking the possibility of its application in industry or the development of functional foods.

Improvement of water transport and carotenoid retention during drying of papaya by applying ultrasonic osmotic pretreatment

The effect of ultrasound and osmotic dehydration pretreatments on papaya drying kinetics was investigated. The ultrasound pretreatment was carried out in an ultrasonic bath at 30 A degrees C. The osmotic pretreatment in sucrose solution was carried out in an incubator at 34 A degrees C and agitation of 80 rpm for 210 min. The drying process was conducted in a fixed bed dryer at 70 A degrees C. Experimental data were fitted successfully using the Page model for dried fresh and pretreated fruits, with coefficient of determination greater than 0.9992 and average relative error lower that 14.4 %. The diffusional model was used to describe the moisture transfer, and the effective water diffusivity was identified in the order of 10(-9) m(2) s(-1). It was found that drying rates of osmosed fruits were the lowest due to the presence of infused solutes, while the ultrasound pretreatment contributed to faster drying rates. Evaluation of the dried fruit was performed by means of total carotenoids retention. Ultrasound treatments in distilled water prior to air-drying gave rise to dried papayas with retention of carotenoids in the range 30.4-39.8 % and the ultrasonic-assisted osmotic dehydration of papayas showed carotenoids retention values up to 64.9 %, whereas the dried fruit without pretreatment showed carotenoids retention lower than 24 %.

Effect of sibling competition and male carotenoid supply on offspring condition and oxidative stress

Early developmental conditions have major implications for an individual's fitness. In species where offspring are born simultaneously, the level of sibling competition for food access is intense. In birds, high sibling competition may subject nestlings to decreased growth rate as a result of limited food and increased levels of oxidative stress through high metabolic activity induced by begging behaviors. We manipulated the level of sibling competition in a natural population of great tits and assessed the consequences for nestling body condition and resistance to oxidative stress. In a full factorial design, we both augmented brood size to increase sibling competition and supplemented the male parents with physiological doses of carotenoids thereby doubling the natural carotenoid intake, aiming at increasing the males' investment in current reproduction and thereby decreasing sibling competition. Nestling body mass was reduced by the brood enlargement and enhanced by the carotenoid supplementation of fathers. Nestling resistance to oxidative stress, measured as total antioxidant defenses in whole blood, was not influenced by the treatments. Because nestlings experience high metabolic activities, an absence of an effect of sibling competition on free radicals production seems unlikely. Nestling body mass decreased and resistance to oxidative stress tended to increase with initial brood size, and hence these correlational effects suggest a trade-off between morphological growth and development of the antioxidant system. However, the result of the experimental treatment did not support this trade-off hypothesis. Alternatively, it suggests that nestling developed compensatory mechanisms that were not detected by our antioxidant capacity measure.

Oxidative damage to DNA related to survivorship and carotenoid-based sexual ornamentation in the common yellowthroat

Carotenoid-based sexual ornaments are hypothesized to be reliable signals of male quality, based on an allocation trade-off between the use of carotenoids as pigments and their use in antioxidant defence against reactive oxygen species. Carotenoids appear to be poor antioxidants in vivo, however, and it is not clear whether variation in ornament expression is correlated with measures of oxidative stress (OXS) under natural conditions. We used single-cell gel electrophoresis to assay oxidative damage to erythrocyte DNA in the common yellowthroat (Geothlypis trichas), a sexually dichromatic warbler in which sexual selection favours components of the males’ yellow ‘bib’. We found that the level of DNA damage sustained by males predicted their overwinter survivorship and was reflected in the quality of their plumage. Males with brighter yellow bibs showed lower levels of DNA damage, both during the year the plumage was sampled (such that yellow brightness signalled current OXS) and during the previous year (such that yellow brightness signalled past OXS). We suggest that carotenoid-based ornaments can convey information about OXS to prospective mates and that further work exploring the proximate mechanism(s) linking OXS to coloration is warranted.