Individual Carotenoid Concentrations Research Articles

Inclusion of Lutein and Zeaxanthin in Standard or Omega-3 Fatty Acids Enriched Laying Hens Diets: Effects on Performance, Egg Quality Parameters, Fatty Acid Composition and Individual Carotenoids Concentration of Egg Yolk

The effects of lutein (L) and zeaxanthin (Z) inclusion to standard or omega-3 fatty acids enriched laying hens’ diets, on performance, egg quality parameters, fatty acids and individual carotenoids of egg yolk was investigated. The dietary treatments were designed as: standard (S) diet, 50 mg/kg L plus 50 mg/kg Z (S+L/Z (50+50)) or 100 mg/kg lutein plus 100 mg/kg zeaxanthin (S+L/Z (100+100)) added to S diet, Omega-3 fatty acids enriched (OM3) diet, 50 mg/kg lutein plus 50 mg/kg zeaxanthin (OM3+L/Z (50+50) or 100 mg/kg lutein plus 100 mg/kg zeaxanthin (OM3+L/Z (100+100)) added to OM3 diet. Hens with a mean live weight of 1525.88 ± 31.19 g were divided into six treatment groups. During the experimental period, there was not any mortality rate recorded. Dietary inclusion of L/Z (50+50 or 100+100) either in S or OM3 diets did not alter performance parameters. Eggs from hens fed enriched diet (OM3) significantly increased (P<0.05) the albumen index and Haugh unit. The type of dietary fat source (F), equal dose of L/Z or F×L/Z on egg yolk lutein, unknown carotenoids and total carotenoid concentration were found to be significant. However, the F on zeaxanthin concentration was not significant (P>0.05). The yolk color a* value was significantly increased by inclusion of L/Z (50+50 or 100+100) to diets (S or OM3) (P<0.05) and this was correlated with increased (P<0.05) concentration of individual carotenoids. The major changes in egg yolk fatty acid MUFA and PUFA composition occurred when laying hen fed with OM3 diet. Key words: Egg yolk, omega-3, carotenoids, lutein, zeaxanth

Characterizing carotenoids in cyanobacterial cultures – Opportunities and implications for paleolimnological studies

Cyanobacterial blooms are increasing in frequency and intensity globally, impacting lake ecosystem health and posing a risk to human and animal health due to the toxins they can produce. Cyanobacterial pigments preserved in lake sediments provide a useful means of understanding the changes that have led to cyanobacterial blooms in lakes. However, there is some uncertainty as to whether specific carotenoids are unique to certain genera or types of cyanobacteria. To fill this knowledge gap, we analyzed pigments in 34 cyanobacteria cultures and applied the findings to sediments from three New Zealand lakes. The cyanobacterial carotenoids canthaxanthin, echinenone and zeaxanthin were detected in all cultures, whereas myxoxanthophyll was only detected in ten cultures (Microcoleus, Planktothrix and the picocyanobacteria cultures; Synechococcaceae). The sum of the individual carotenoid concentrations provided the strongest relationship with cyanobacterial biomass (R2 = 0.58) and could be used in paleolimnology studies to evaluate general cyanobacterial abundance. Ratios of canthaxanthin, zeaxanthin and myxoxanthophyll relative to echinenone indicated that carotenoid ratios could be used to differentiate picocyanobacteria and bloom-forming cyanobacteria, to some degree. High zeaxanthin/echinenone ratios were measured in picocyanobacteria and low zeaxanthin/echinenone ratios were measured in bloom-forming cyanobacteria. The zeaxanthin/echinenone ratio was applied to sediment core samples where the cyanobacterial community was also evaluated by 16S rRNA gene metabarcoding, with the zeaxanthin/echinenone ratios showing similar patterns to those observed in the cultures. The preliminary assessment described here suggests that zeaxanthin/echinenone ratios could provide a valuable paleoecological proxy for evaluating historical shifts in cyanobacterial communities and warrants further exploration.

New associations of serum β-carotene, lycopene, and zeaxanthin concentrations with NR1H3, APOB, RDH12, AND CYP genes.

Variation in carotenoid bioavailability at individual and population levels might depend on host‐related factors where genetic variation has a part to play. It manifests itself through the proteins involved in carotenoid intestinal absorption and metabolism, blood lipoprotein transport, or tissue uptake. This study aims to identify novel SNPs which could be associated with carotenoid serum concentrations. A total of 265 self‐reported healthy individuals of Lithuanian origin were genotyped (Illumina HumanOmniExpress‐12v1.0 or v1.1 and Infinium OmniExpress‐24v1.2 arrays) and fasting blood serum concentrations of β‐ and α‐carotene, β‐cryptoxanthin, lycopene, lutein, and zeaxanthin were measured (Shimadzu Prominence HPLC system). According to the individual carotenoid concentrations, the cohort was subdivided into quartiles. Q1 and Q4 were used for the following association analysis. The set of 2883 SNPs in 109 potential candidate genes (assumed for a direct or indirect role in carotenoid bioavailability) was analyzed. Liver X receptor alpha (NR1H3) “transport” polymorphisms rs2279238 (p = 2.129 × 10−5) and rs11039155 (p = 2.984 × 10−5), and apolipoprotein B (APOB) “transport” polymorphism rs550619 (p = 4.844 × 10−5) were associated with higher zeaxanthin concentration. Retinol dehydrogenase 12 (RDH12) “functional partner” polymorphism rs756473 (p = 7.422 × 10−5) was associated with higher lycopene concentration. Twenty‐one cytochrome P450 (CYP2C9, CYP2C18, and CYP2C19) “metabolism” polymorphisms in locus 10q23.33 were significantly associated with higher β‐carotene concentration. To conclude, four novel genomic loci were found to be associated with carotenoid serum levels. Zeaxanthin, lycopene, and β‐carotene serum concentrations might depend on genetic variation in NR1H3, APOB, RDH12 and CYP2C9, CYP2C18, and CYP2C19 genes.

Serum Carotenoids Reveal Poor Fruit and Vegetable Intake among Schoolchildren in Burkina Faso.

The health benefits of fruits and vegetables are well-documented. Those rich in provitamin A carotenoids are good sources of vitamin A. This cross-sectional study indirectly assessed fruit and vegetable intakes using serum carotenoids in 193 schoolchildren aged 7 to 12 years in the Western part of Burkina Faso. The mean total serum carotenoid concentration was 0.23 ± 0.29 µmol/L, which included α- and β-carotene, lutein, and β-cryptoxanthin, and determined with serum retinol concentrations in a single analysis with high performance liquid chromatography. Serum retinol concentration was 0.80 ± 0.35 µmol/L with 46% of children (n = 88) having low values <0.7 µmol/L. Total serum carotene (the sum of α- and β-carotene) concentration was 0.13 ± 0.24 µmol/L, well below the reference range of 0.9–3.7 µmol carotene/L used to assess habitual intake of fruits and vegetables. Individual carotenoid concentrations were determined for α-carotene (0.01 ± 0.05 µmol/L), β-carotene (0.17 ± 0.24 µmol/L), β-cryptoxanthin (0.07 ± 0.06 µmol/L), and lutein (0.06 ± 0.05 µmol/L). These results confirm the previously measured high prevalence of low serum vitamin A concentrations and adds information about low serum carotenoids among schoolchildren suggesting that they have low intakes of provitamin A-rich fruits and vegetables.

UV radiation promotes flavonoid biosynthesis, while negatively affecting the biosynthesis and the de-epoxidation of xanthophylls: Consequence for photoprotection?

There is evidence that UV radiation may detrimentally affect the biosynthesis of carotenoids, particularly de-epoxided xanthophylls, while strongly promoting phenylpropanoid, particularly flavonoid biosynthesis in a range of taxa. Here we tested the hypothesis that mesophyll flavonoids might protect chloroplasts from UV-induced photo-oxidative damage, by partially compensating for the UV-induced depression of xanthophyll biosynthesis. To test this hypothesis we grew two members of the Oleaceae family, Ligustrum vulgare L. and Phillyrea latifolia L., under either partial shading or fully exposed to sunlight, in the presence or in the absence of UV radiation. The examined species, which display very similar flavonoid composition, largely differ in their ability to limit the transmission of UV and visible light through the leaf and, hence, in the accumulation of flavonoids in mesophyll cells. We conducted measurements of photosynthesis, chlorophyll a fluorescence kinetics, the concentrations of individual carotenoids and phenylpropanoids at the level of whole-leaf, as well as the content of epidermal flavonoids. We also performed multispectral fluorescence micro-imaging to unveil the intra-cellular distribution of flavonoids in mesophyll cells. UV radiation decreased the concentration of carotenoids, particularly of xanthophylls, while greatly promoting the accumulation of flavonoids in palisade parenchyma cells. These effects were much greater in L. vulgare than in P. latifolia. UV radiation significantly inhibited the de-epoxidation of xanthophyll cycle pigments, while enhancing the concentration of luteolin, and particularly of quercetin glycosides. Flavonoids accumulated in the vacuole and the chloroplasts in palisade cells proximal to the adaxial epidermis. We hypothesize that flavonoids might complement the photo-protective functions of xanthophylls in the chloroplasts of mesophyll cells exposed to the greatest doses of UV radiation. However, UV radiation might result in adaxial mesophyll cells being less effective in dissipating the excess of radiant energy, e.g., by decreasing their capacity of thermal dissipation of excess visible light in the chloroplast.

Salinity stress constrains photosynthesis in Fraxinus ornus more when growing in partial shading than in full sunlight: consequences for the antioxidant defence system.

A major challenge in plant ecophysiology is understanding the effects of multiple sub-optimal environmental conditions on plant performance. In most Mediterranean areas soil salinity builds up during the summer because of low availability of soil water coupled with hot temperatures. Although sunlight and soil salinity may strongly interact in determining a plant's performance, this has received relatively little attention. Two-year-old seedlings of Fraxinus ornus were grown outdoors in pots during a Mediterranean summer in either 45 % (shaded plants) or 100 % (sun plants) sunlight irradiance and were supplied with either deionized water or deionized water plus 75 mm NaCl. Morpho-anatomical traits, water and ionic relations, gas exchange and photosystem II performance, concentrations of individual carotenoids, activity of antioxidant enzymes, concentrations of ascorbic acid and individual polyphenols were measured in leaves. Leaf oxidative stress and damage were estimated by in vivo analysis of stable free radicals and ultrastructural analyses. Leaf concentrations of potentially toxic ions did not markedly differ in shaded or sun plants in response to salinity. Leaves of sun plants displayed superior water use efficiency compared with leaves of shaded plants, irrespective of salinity treatment, and had both better stomatal control and higher CO2 carboxylation efficiency than leaves of shaded plants. In the salt-treated groups, the adverse effects of excess midday irradiance were greater in shade than in sun plants. The activity of enzymes responsible for detoxifying hydrogen peroxide decreased in shaded plants and increased in sun plants as a result of salinity stress. In contrast, the activity of guaiacol peroxidase and the concentration of phenylpropanoids increased steeply in response to salinity in shaded plants but were unaffected in sun plants. It is concluded that salinity may constrain the performance of plants growing under partial shading more severely than that of plants growing under full sun during summer. The results suggest co-ordination within the antioxidant defence network aimed at detoxifying salt-induced generation of reactive oxygen species.

Stability of Carotenoids, Total Phenolics and In Vitro Antioxidant Capacity in the Thermal Processing of Orange-Fleshed Sweet Potato (Ipomoea batatas Lam.) Cultivars Grown in Brazil

Intervention strategies regarding the biofortification of orange-fleshed sweet potato, which is a rich source of carotenoids for combating vitamin A deficiency, are being developed in Brazil. This study was conducted to evaluate the concentrations of individual carotenoids, total phenolic compounds and antioxidant capacity in the roots of four biofortified sweet potato cultivars that were raw or processed by four common heat treatments. HPLC, Folin-Ciocalteu, DPPH and ABTS assays were used. All cultivars showed high levels of carotenoids in raw roots, predominantly all-trans-β-carotene (79.1-128.5mg.100g(-1) DW), suggesting a high estimated vitamin A activity. The CNPH 1194 cultivar reported carotenoids values highest than those of other cultivars (p < 0.05). The total phenolic compounds varied among cultivars and heat treatments (0.96-2.05mg.g(-1) DW). In most cases, the heat treatments resulted in a significant decrease in the carotenoids and phenolic compounds contents as well as antioxidant capacity. Processing of flour presented the greatest losses of major carotenoids and phenolics. The phenolic compounds showed more stability than carotenoids after processing. There were significant correlations between the carotenoids and phenolic compounds and the antioxidant capacity.

The genes and enzymes of the carotenoid metabolic pathway in Vitis vinifera L.

BackgroundCarotenoids are a heterogeneous group of plant isoprenoids primarily involved in photosynthesis. In plants the cleavage of carotenoids leads to the formation of the phytohormones abscisic acid and strigolactone, and C13-norisoprenoids involved in the characteristic flavour and aroma compounds in flowers and fruits and are of specific importance in the varietal character of grapes and wine. This work extends the previous reports of carotenoid gene expression and photosynthetic pigment analysis by providing an up-to-date pathway analysis and an important framework for the analysis of carotenoid metabolic pathways in grapevine.ResultsComparative genomics was used to identify 42 genes putatively involved in carotenoid biosynthesis/catabolism in grapevine. The genes are distributed on 16 of the 19 chromosomes and have been localised to the physical map of the heterozygous ENTAV115 grapevine sequence. Nine of the genes occur as single copies whereas the rest of the carotenoid metabolic genes have more than one paralogue. The cDNA copies of eleven corresponding genes from Vitis vinifera L. cv. Pinotage were characterised, and four where shown to be functional. Microarrays provided expression profiles of 39 accessions in the metabolic pathway during three berry developmental stages in Sauvignon blanc, whereas an optimised HPLC analysis provided the concentrations of individual carotenoids. This provides evidence of the functioning of the lutein epoxide cycle and the respective genes in grapevine. Similarly, orthologues of genes leading to the formation of strigolactone involved in shoot branching inhibition were identified: CCD7, CCD8 and MAX1. Moreover, the isoforms typically have different expression patterns, confirming the complex regulation of the pathway. Of particular interest is the expression pattern of the three VvNCEDs: Our results support previous findings that VvNCED3 is likely the isoform linked to ABA content in berries.ConclusionsThe carotenoid metabolic pathway is well characterised, and the genes and enzymes have been studied in a number of plants. The study of the 42 carotenoid pathway genes of grapevine showed that they share a high degree of similarity with other eudicots. Expression and pigment profiling of developing berries provided insights into the most complete grapevine carotenoid pathway representation. This study represents an important reference study for further characterisation of carotenoid biosynthesis and catabolism in grapevine.

Photoprotection by carotenoid pigments in the yeast Rhodotorula mucilaginosa: the role of torularhodin

In this paper we report the relationship between carotenoids and ergosterol and cell UV-B resistance in different strains of the yeast Rhodotorula mucilaginosa. Cell survival was studied using a set of 13 strains; additionally, two mutants (a hyper-producing one and a colourless one) in combination with diphenylamine (DPA), a carotenogenesis inhibitor, were used. A positive correlation between total carotenoids and survival to UV-B radiation was found. However, when individual carotenoid concentrations were tested, only torularhodin was found to be significantly related to UV-B survival. On the contrary, ergosterol did not affect survival. The hyper-pigmented strain showed an enhanced survival (up to 250%) compared to the parental strain, while the survival of the albino mutant was similar to that experienced by the parental strain; however, observed changes in survival were dose dependent. The cyclobutane pyrimidine dimers (CPDs), one of the major forms of DNA damage caused by UV exposure, appears as unrelated to the accumulation of carotenoids and cell survival. These results indicate that bearing higher torularhodin concentrations enhances UV-B survival in yeasts and, thus, the accumulation of this pigment constitutes an important mechanism that improves the resistance of yeasts to UV-B.

Total and individual carotenoid profiles in Solanum phureja cultivated potatoes: II. Development and application of near-infrared reflectance spectroscopy (NIRS) calibrations for germplasm characterization

Calibration equations to estimate total and individual carotenoid concentrations by near-infrared reflectance spectroscopy (NIRS) in Solanum phureja potato germplasm were developed, validated and applied to the characterization of 152 S. phureja germplasm accessions. The cross, external and independent validation of the calibrations indicate that total carotenoids and zeaxanthin concentrations can be estimated by NIRS with high accuracy and that NIRS can be used to differentiate accessions with low, medium and high concentrations of violaxanthin, antheraxanthin, lutein or β-carotene. Significant variability was revealed for total and all individual carotenoids measured. Two varieties with significant zeaxanthin concentrations (above 1000 μg 100 g −1 FW) and a group of 43 accessions with relatively high β-carotene concentrations (above 10 μg 100 g −1 FW) were identified. The positive relation between total carotenoids and zeaxanthin as well as their negative relation with β-carotene reported for 23 accessions in the companion manuscript was upheld. The use of NIRS in estimating and characterizing a large germplasm collection has permitted the identification of accessions with high total carotenoids and zeaxanthin and relatively high β-carotene concentrations at low cost and in reduced time compared to HPLC. This can be applied to documentation of descriptive and potentially beneficial characteristics of potato genebank materials, the assessment of the food value of present varieties, and the selection of parents in a breeding program oriented to nutritional enhancement of potato.

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 spectrophotometry 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 β-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 μg 100 g −1 FW) comprised largely of zeaxanthin (658–1290 μg 100 g −1 FW) with very low or no presence of β-carotene (below 5.4 μg 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 μg 100 g −1 FW) and very low or no zeaxanthin, with lutein and violaxanthin as the predominant carotenoids and relatively high concentrations of β-carotene (up to 27 μg 100 g −1 FW). Five accessions with significant concentrations of zeaxanthin were identified with the accession 703566 showing the highest concentration (1290 μg 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 β-carotene concentration.

Persistence of carotenoid pigments in the blood of concentrate-finished grazing sheep: its significance for the traceability of grass-feeding.

Carotenoid pigments are good biomarkers of grass feeding in sheep. However, grazing lambs are often stall-finished because of grass shortage. We investigated the nature of the carotenoids present in sheep blood and their persistence in this tissue. Four treatments were compared: 1) feeding a concentrate-based diet (n = 10 lambs), 2) grazing followed by a long stall-finishing period (n = 10), 3) grazing followed by a short stall-finishing period (n = 10), and 4) grazing to slaughter weight (n = 10). The concentrate supply was regulated to have similar average daily gain for all treatments. The 40 lambs were allocated to either the grazing or the stall treatments on the basis of their birth date, birth weight, and body weight. The 30 grazing lambs were further allocated to long-stall, short-stall, or grass treatment on the basis of their body weight and plasma carotenoid content. Plasma content of total carotenoids was measured by spectrophotometry during the grazing and the stall periods for all lambs and at slaughter weight for the eight heaviest lambs of each treatment. Analysis of the nature and the concentration of individual carotenoids was performed by HPLC on pasture and stall diets and on blood of grazing lambs. The carotenoid content of the stall diet was 2 to 3% that of the pasture diet. Lutein, zeaxanthin, and beta-carotene accounted for 43 to 58%, 3 to 17%, and 0 to 7% of total plasma carotenoids in grazing lambs, respectively. Two unknown polar carotenoids, expressed in lutein equivalent, accounted for 10 to 22% and 0 to 9% of total carotenoids. Plasma carotenoid content during the grazing and the finishing periods varied among animals (P < 0.001). At slaughter weight, plasma carotenoid content was higher for grass-fed than for stall-fed, long-stall finished, or short-stall finished lambs (P < 0.001), and reliably distinguished grass-fed lambs from all the others. Plasma carotenoid content decreased exponentially with the interval from starting on the stall diet (P < 0.005). The deceleration parameter of the model increased linearly with lamb average daily gain during the stall-finishing period, suggesting that the turnover of carotenoids in the blood may depend on the level of intake of the stall-finishing diet. After 4 to 13 d on the stall diet, depending on the initial plasma carotenoid concentration, plasma carotenoid concentration of previously grazed, stall-finished lambs fell to the values of lambs fed a concentrate diet without grazing. Such a low persistence is of interest for discriminating grazing lambs from stall-finished grazing lambs.

Individual carotenoid concentrations in adipose tissue and plasma as biomarkers of dietary intake , ,

Plasma and adipose tissue concentrations of carotenoids are thought to reflect short- and long-term intakes of carotenoids, respectively. The ability of adipose tissue carotenoid concentrations to reflect dietary intake in population studies is unknown. We examined the relation between intakes of the major dietary carotenoids and their concentrations in plasma and adipose tissue. A blood sample and an adipose tissue biopsy sample were collected from 115 women and 344 men in Costa Rica after they had fasted overnight, and a dietary interview based on a 135-item food-frequency questionnaire was administered. After carotenoid intake was adjusted for total energy intake and plasma concentrations were adjusted for HDL-, LDL-, and VLDL-cholesterol concentrations, we calculated partial Spearman correlation coefficients that were adjusted for age, sex, smoking, and body mass index. In women, the correlations (r) between intakes and concentrations of alpha-carotene, beta-carotene, beta-cryptoxanthin, and lutein+zeaxanthin were 0.25, 0.29, 0.44, and 0.17, respectively (P < 0.05 for r > or = 0.19), in adipose tissue and 0.26, 0.13, 0.55, and 0.22 in plasma. In men, these values were 0.04, 0.07, 0.23, and 0.06 in adipose tissue and 0.24, 0.22, 0.44, and 0.20 in plasma. In women and men, correlations for lycopene were higher in plasma (r = 0.19 and 0.35, respectively) than in adipose tissue (r = 0.14 and 0.26). The relative abundance of each carotenoid in the diet was similar to its distribution in plasma but not in adipose tissue. The usefulness of adipose tissue and plasma carotenoids as biomarkers of intake is similar, although correlations for individual carotenoids vary substantially.

Serum carotenoids and tocopherols and incidence of age-related nuclear cataract

It is not known whether the protective effects of antioxidants on cataract observed in experimental animals are relevant to age-related opacities in humans. The relations of serum carotenoids and tocopherols to the incidence of age-related nuclear cataract were investigated in a random sample of 400 adults, 50-86 y of age, in the Beaver Dam Eye Study. Nuclear opacity was assessed by using lens photographs taken at baseline (in 1988-1990) and follow-up (in 1993-1995). Nonfasting concentrations of individual carotenoids and alpha- and gamma-tocopherol, were determined from serum obtained at baseline. A total of 252 persons were eligible for incident cataract, of whom 57 developed nuclear cataract in at least one eye. Results were adjusted for age, smoking, serum cholesterol, heavy drinking, adiposity, and, in the tocopherol models, dietary linoleic acid intake. Only serum tocopherol (the sum of alpha- and gamma-tocopherol, in micromol/mmol cholesterol) was associated with cataract. For total serum tocopherol, persons in tertile 3 had a lower risk of cataract than persons in tertile 1 [odds ratio (OR): 0.4; 95% CI: 0.2, 0.9; P = 0.03 for linear trend]. Although serum carotenoids were not significantly associated with nuclear cataract, marginal inverse associations with lutein (OR: 0.3; 95% CI: 0.1, 1.2; P = 0.13 for linear trend) and cryptoxanthin (OR: 0.3; 95% CI: 0.1, 1.3; P = 0.11 for linear trend) were suggested in people < or = 65 y of age. Findings were compatible with the possibility that nuclear cataract may be linked inversely to vitamin E status, but neither strongly supported nor negated the hypothesized inverse association of nuclear cataract with serum carotenoids.

Effect of menstrual cycle phase on the concentration of individual carotenoids in lipoproteins of premenopausal women: a controlled dietary study

Because premenopausal women experience cyclic fluctuations of plasma carotenoids and their lipoprotein carriers, it is hypothesized that carotenoid concentrations in lipoprotein fractions fluctuate by phase of the menstrual cycle. Nine women ate a standard set of carotenoid-rich foods daily for two cycles under isoenergetic conditions. In the second cycle, hormones and carotenoids in lipoprotein fractions were measured in the early and late follicular and luteal phases. alpha-Carotene concentrations in the LDL fraction were lower in the early than in the late follicular phase (P = 0.03) on the basis of regression analysis. beta-carotene concentrations in the LDL fraction and the HDL2 subfraction were higher in the late follicular than in the luteal phase (P = 0.02 and P = 0.04, respectively). Lutein/zeaxanthin concentrations in the LDL and HDL fractions were higher in the late follicular than in the luteal phase (P = 0.03 and P = 0.02, respectively). In each phase, 80% of alpha-carotene, 82% of beta-carotene, 85% of lycopene, and 64% of lutein/zeaxanthin were distributed in the LDL fraction. Among the hydrocarbon cartenoids, 18% of alpha-carotene and of beta-carotene and 13% of lycopene were distributed in the HDL fraction, with slightly more in the HDL2 than in the HDL3 subfraction. In contrast 34% of lutein/zeaxanthin was distributed in the HDL fraction with more concentrated in the HDL3 than in the HDL2 subfraction. Less than 4% of any carotenoid was found in the VLDL + IDL (intermediate-density-lipoprotein) fractions. Thus, the hydrocarbon carotenoids were highly concentrated in the LDL fraction and xanthophyll was more evenly distributed in the LDL and HDL fractions. The cyclic fluctuations of these carotenoids in lipoprotein fractions add another dimension to the understanding of their transport and physiologic function.

Effects of storage and handling conditions on concentrations of individual carotenoids, retinol, and tocopherol in plasma.

We investigated the effects of storage and handling on measured values for carotenoids, retinol, and tocopherol in plasma. We found no significant differences in the concentrations of these analytes measured in plasma samples that were frozen immediately after separation as compared with replicate samples maintained at room temperature in the dark for 24 h. Analytes were stable in solvents for at least 18 h at 23 degrees C after extraction. Purging samples with nitrogen gas before freezing had no detectable beneficial effects. All analytes were stable in plasma stored at -70 degrees C for at least 28 months or at -20 degrees C for five months. By 15 months the concentrations of carotenoids were significantly less (P less than 0.05) in plasma stored at -20 degrees C than in plasma stored at -70 degrees C, while retinol and tocopherol concentrations were not significantly different. Concomitant with the decrease in carotenoids was the appearance of unidentified peaks in the ultraviolet. Adding ascorbic acid or butylated hydroxytoluene antioxidants to the precipitating solvent did not alter the losses of carotenoids or alter the appearance of unidentified peaks. Under appropriate conditions, plasma carotenoids, retinol, and tocopherol are stable for more than two years.

Carotenoid and tocopherol composition of human adipose tissue

Concentrations of individual carotenoids and tocopherols were determined in abdominal adipose tissue from 19 adults undergoing corrective surgery. Samples were extracted and saponified before separation and quantitation of carotenoids and tocopherols by reverse-phase high-performance liquid chromatography. Total carotenoid concentration varied 40-fold between individuals, from 0.34 to 13.51 micrograms/g adipose tissue. Beta-carotene and lycopene were the predominant carotenoids, averaging 20.2 and 18.5% of total carotenoids, respectively. In 10 of 19 subjects, the lycopene concentration exceeded that of beta-carotene. Total tocopherol concentrations varied 11-fold, with alpha-tocopherol representing 80.6 +/- 8.1% of the total. Absolute concentrations of both carotenoids and tocopherols were more variable than their relative concentrations. Both beta-carotene and lycopene concentrations were highly correlated with total carotenoid content but there was no correlation between beta-carotene and lycopene or between beta-carotene and alpha-tocopherol concentrations.

Changes in chemical composition of thylakoid membranes during greening of the y-1 mutant of Chlamydomonas reinhardi.

Two fractions of thylakoid membranes (TMF) have been isolated from disrupted (French press) algal cells by using a discontinuous sucrose gradient. TMF-II consists mostly of thylakoid membranes still partially organized in grana; it contains also fragments of chloroplast envelope, pyrenoid tubules, and starch granules; thus it amounts to a fraction of chloroplast fragments which have lost practically all matrix components. TMF-I consists of smaller chloroplast fragments and is contaminated to a larger extent than TMF-II by other subcellular components, primarily mitochondria. TMF-II accounts for about 12% of the protein and 30% of the chlorophyll of the whole cell; it contains cytochrome 554 and carotenoids in the same ratio to chlorophyll as the latter, and shows photosystems I and II activities but lacks enzymatic activities characteristic of the dark reactions. During the greening of the y-1 mutant of Chlamydomonas, TMF's have been isolated over a range of chlorophyll concentrations from 5 to 25 microg/10(7) cells. The results showed that during this period the ratios of chlorophyll to cytochrome 554 and of chlorophyll to carotenoids, and the relative concentrations of individual carotenoids were continuously changing. The findings support the view that during greening, thylakoid membranes are produced by multistep assembly.