Assessment of Astaxanthin Accumulation in Hepatocytes of Atlantic Salmon Fed Different Diets Using NMR Spectroscopy.

This study aimed to assess the astaxanthin (Ax) accumulation in hepatocytes isolated from farmed Atlantic salmon fed different diets (rich marine, poor, poor with marine phospholipids (MPL) and poor with docosahexaenoic acid (DHA)). Nuclear magnetic resonance (NMR) spectroscopy was used for the Ax detection and quantification. The use of the 13C-enriched Ax allowed the assessment of short-time Ax metabolism. The substitution of fish oil and meal in fish feed on plant analogs and the addition of MPL caused further catabolism and decrease of Ax accumulation in hepatocytes from 17 to about 6 mg/kg or to almost zero in the case of DHA addition. Signals assignment of the native and 13C-enriched astaxanthin in acetone were performed using 1D and 2D NMR spectra.

A novel strategy for isolation and purification of fucoxanthinol and fucoxanthin from the diatom Nitzschia laevis.

In this study, the microalga Nitzschia laevis (N. laevis) can accumulate a marine carotenoid fucoxanthinol. In particular, fucoxanthinol was firstly isolated from microalgae, accompanied by its derivative fucoxanthin. The identification and quantification of fucoxanthinol and fucoxanthin were determined by ultra-performance liquid chromatography coupled to photodiode array detector-quadrupole/travelling-wave ion mobility mass spectrometry/time-of-flight mass spectrometry (UPLC-PDA-TWIMS-QTOF-MS). Furthermore, a cost-effective approach mediated with solid-phase extraction (SPE) and thin-layer chromatography (TLC) technique was used to isolate and purify fucoxanthinol and fucoxanthin from the extracts of N. laevis. This two-step method can obtain 98% fucoxanthinol and 95% fucoxanthin, with the recovery efficiencies of around 85% for fucoxanthinol and 70% for fucoxanthin, respectively. Moreover, 1H and 13C nuclear magnetic resonance (NMR) techniques were adopted to record the purified compounds for supporting the above results. In all, the developed method has a promising potential to purify fucoxanthinol and fucoxanthin of microalgae for food and pharmaceutical applications.

Health condition assessment for vegetation exposed to heavy metal pollution through airborne hyperspectral data.

Recent advancements in hyperspectral remote sensing technology now provide improved diagnostic capabilities to assess vegetation health conditions. This paper uses a set of 13 vegetation health indices related to chlorophyll, xanthophyll, blue/green/red ratio and structure from airborne hyperspectral reflectance data collected around a derelict mining area in Yerranderie, New South Wales, Australia. The studied area has ten historic mine shafts with a legacy of heavy metals and acidic contamination in a pristine ecosystem now recognised as Great Blue Mountain World Heritage Area. The forest is predominantly comprised of different species of Eucalyptus trees. In addition to the airborne survey, ground-based spectra of the tree leaves were collected along the two accessible heavy metal contaminated pathways. The stream networks in the area were classified and the geospatial patterns of vegetation health were analysed along the Tonalli River, a major water tributary flowing through the National Park. Despite the inflow of contaminated water from the near-mine streams, the measured vegetation health indices along Tonalli River were found to remain unchanged. The responses of the vegetation health indices between the near-mine and away-mine streams were found similar. Based on the along-stream and inter-stream analysis of the spectral indices of vegetation health, no significant impact of the heavy metal pollution could be noticed. The results indicate the possibility of the vegetation having developed immunity towards the high levels of heavy metal pollution over a century of exposure.

Assessment of antioxidant activity in Victorian marine algal extracts using high performance thin-layer chromatography and multivariate analysis.

The aim of this study was to develop and validate a rapid and simple high performance thin layer chromatographic (HPTLC) method to screen for antioxidant activity in algal samples. 16 algal species were collected from local Victorian beaches. Fucoxanthin, one of the most abundant marine carotenoids was quantified directly from the HPTLC plates before derivatization, while derivatization either with 2,2-diphenyl-1-picrylhydrazyl (DPPH) or ferric chloride (FeCl3) was used to analyze antioxidants in marine algae, based on their ability to scavenge non biological stable free radical (DPPH) or to chelate iron ions. Principal component analysis of obtained HPTLC fingerprints has classified algae species into 5 groups according to their chemical/antioxidant profiles. The investigated brown algae samples were found to be rich in non-and moderate-polar compounds and phenolic compounds with antioxidant activity. Most of the phenolic iron chelators also have shown free radical scavenging activity. Strong positive and significant correlations between total phenolic content and DPPH radical scavenging activity showed that, phenolic compounds, including flavonoids are the main contributors of antioxidant activity in these species. The results suggest that certain brown algae possess significantly higher antioxidant potential when compared to red or green algae and could be considered for future applications in medicine, dietary supplements, cosmetics or food industries. Cystophora monilifera extract was found to have the highest antioxidant concentration, followed by Zonaria angustata, Cystophora pectinate, Codium fragile, and Cystophora pectinata. Fucoxanthin was found mainly in the brown algae species. The proposed methods provide an edge in terms of screening for antioxidants and quantification of antioxidant constituents in complex mixtures. The current application also demonstrates flexibility and versatility of a standard HPTLC system in the drug discovery. Proposed methods could be used for the bioassay-guided isolation of unknown natural antioxidants and subsequent identification if combined with spectroscopic identification.

Assessment of tissue distribution and concentration of ß-cryptoxanthin in response to varying amounts of dietary ß-cryptoxanthin in the Mongolian gerbil.

There is a general lack of knowledge regarding the absorption and tissue storage of the provitamin A carotenoid ß-cryptoxanthin. The present study investigated the whole-body tissue distribution of ß-cryptoxanthin in an appropriate small animal model, the Mongolian gerbil (Meriones unguiculatus), for human provitamin A carotenoid metabolism. After 5 d of carotenoid depletion, five gerbils were euthanised for baseline measurements. The remaining gerbils were placed in three weight-matched treatment groups (n 8). All the groups received 20 µg/d of ß-cryptoxanthin from tangerine concentrate, while the second and third groups received an additional 20 and 40 µg/d of pure ß-cryptoxanthin (CX40 and CX60), respectively, for 21 d. During the last 2 d of the study, urine and faecal samples of two gerbils from each treatment group were collected. ß-Cryptoxanthin was detected in the whole blood, and in twelve of the fourteen tissues analysed. Most tissues resembled the liver, in which the concentrations of ß-cryptoxanthin were significantly higher in the CX60 (17·8 (sem 0·7) µg/organ; P= 0·004) and CX40 (16·2 (sem 0·9) µg/organ; P= 0·006) groups than in the CX20 group (13·3 (sem 0·4) µg/organ). However, in intestinal tissues, the concentrations of ß-cryptoxanthin increased only in the CX60 group. Despite elevated vitamin A concentrations in tissues at baseline due to pre-study diets containing high levels of vitamin A, ß-cryptoxanthin maintained those vitamin A stores. These results indicate that ß-cryptoxanthin is stored in many tissues, potentially suggesting that its functions are widespread.

In vitro assessment of the bioaccessibility of carotenoids from sun-dried chilli peppers.

Chilli peppers have been recognized as an excellent source of antioxidants as they are rich in bioactive phytochemicals such as carotenoids which are known to exert various beneficial effects in vivo. Absorption is an important factor in the determination of the potential biological effects of carotenoids. The bioaccessibility of a food constituent such as a carotenoid represents its potential to be absorbed in humans. There is very limited information in the literature regarding the content and bioaccessibility of carotenoids from dried peppers. Therefore, the objectives of the present study were: first, to determine the carotenoid content of 20 varieties of red, orange or yellow coloured sun-dried chilli peppers belonging to either of four Capsicum species (annuum, baccatum, chinense and chacoense); and second, to quantify the carotenoid micellarization (bioaccessibility) following an in vitro digestion procedure. Red peppers had a higher carotenoid content and bioaccessibility than either the orange peppers or yellow pepper. Xanthophylls showed greater bioaccessibility than carotenes. Our findings confirm that dried chilli peppers are a good source of carotenoids.

Astaxanthin and canthaxanthin (xanthophyll) as supplements in rainbow trout diet: in vivo assessment of residual levels and contributions to human health.

Many studies have demonstrated that xanthophylls, such as astaxanthin, have beneficial effects in human health, and their use in food supplements is thus encouraged. Moreover, such nutrients are frequently used in aquaculture to meet consumer demand for salmonoid flesh pigmentation. In this study different xanthophyll administration protocols were tested to verify pigmentation properties and safety of such mixtures of additives in trout diet. Residues of xanthophylls in muscle samples were determined by HPLC-MS/MS, reaching levels of 3.70 ± 0.04 mg/kg (astaxanthin) and 1.21 ± 0.06 mg/kg (canthaxanthin) during a 56 day administration period. On the basis of the average fish consumption in the human diet, the highest astaxanthin and canthaxanthin concentrations detected in trout fillets could result in weekly intakes of 1.63 and 0.53 mg, respectively, in humans; these values are not sufficient to achieve the positive effects described by many authors, but their residues could still represent an important source of carotenoids, alternative to the use of synthetic dietary supplements.

Supercritical carbon dioxide extraction of fucoxanthin from Undaria pinnatifida.

Undaria pinnatifida, commonly known as wakame in Japan, is one species of brown seaweeds containing valuable bioactive organic compounds such as fucoxanthin, a carotenoid, which has numerous functional properties. However, most of the seaweeds that do not meet strict quality standards are normally discarded as wastes or returned to the sea, a situation which is becoming an environmental concern. In this research, supercritical carbon dioxide (SCCO2) extraction was investigated for the isolation of fucoxanthin. SCCO2 extraction experiments were carried out at temperature range of 25-60 °C and pressure range of 20-40 MPa, at a carbon dioxide flow rate of 1.0-4.0 mL/min. Results showed that fucoxanthin recovery closed to 80% could be obtained at 40 °C and 40 MPa in extraction time of 180 min. The recovery increased with decreasing temperature and increasing pressure. Pretreatment with microwave (MW) also enhanced the efficiency of extraction due most likely to disruption of the cell membrane. Application of SCCO2, generally regarded as safe and environmentally benign solvent, for extraction of useful bioactive compounds from unwanted or substandard seaweeds look promising in the near future. The extracts obtained using the method can be utilized as food and pharmaceutical additive, and can be used in the development of new health supplements.

Assessment of phytochemical content in human milk during different stages of lactation.

OBJECTIVE: The present study reports the presence of several carotenoids and flavonoids in human milk samples. METHODS: Samples were collected from 17 women who delivered healthy term babies (≥ 37 wk of gestation) at 1-, 4-, and 13-wk postpartum intervals. RESULTS: Epicatechin (63.7-828.5 nmol/L), epicatechin gallate (55.7-645.6 nmol/L), epigallocatechin gallate (215.1-2364.7 nmol/L), naringenin (64.1-722.0 nmol/L), kaempferol (7.8-71.4 nmol/L), hesperetin (74.8-1603.1 nmol/L), and quercetin (32.5-108.6 nmol/L) were present in human milk samples with high inter-/intraindividual variability. With the exception of kaempferol, the mean flavonoid content in human milk was not statistically different among lactation stages. In contrast, carotenoids α-carotene (59.0-23.2 nmol/L), ß-carotene (164.3-88.0 nmol/L), α-cryptoxanthin (30.6-13.5 nmol/L), ß-cryptoxanthin (57.4-24.8 nmol/L), zeaxanthin (46.3-21.4 nmol/L), lutein (121.2-56.4 nmol/L), and lycopene (119.9-49.5 nmol/L) significantly decreased from weeks 1 to 13 of lactation. CONCLUSION: The observed differences in the relative concentrations of the two phytochemical classes in human milk may be a result of several factors, including dietary exposure, stability in the milk matrix, efficiency of absorption/metabolism, and transfer from plasma to human milk. These data support the notion that flavonoids, as with carotenoids, are dietary phytochemicals present in human milk and potentially available to breast-fed infants.

Antiobesity effects of Undaria lipid capsules prepared with scallop phospholipids.

Based on previous research findings, a capsule was developed containing n-3 polyunsaturated fatty acid rich scallop phospholipids (PLs) with an incorporation of brown seaweed (Undaria pinnatifida) lipids (ULs) containing fucoxanthin. The antiobesity effects of the capsules were evaluated with an animal model using 3-wk-old male KK-A(y) mice. Each group received different combinations of lipid (UL, PL, UL + PL, or UL + PL capsule) either incorporated into the diet or into drinking water. Animals were sacrificed after a 4-wk experimental feeding period, and adipose tissues and organs were dissected and weighed. Blood samples were obtained to determine plasma lipid profiles. Uncoupling protein 1 (UCP1) mRNA expression levels were determined by real-time polymerase chain reaction analysis, and UCP1 expression was determined by western blotting analysis. Treatment with either UL alone or UL + PL (capsule) through drinking water resulted in a significant reduction in body weight, compared to the control group. The total white adipose tissue weight of mice fed the UL + PL capsule in drinking water was significantly reduced. Both UCP1 and UCP1 mRNA expression in epididymal fat from mice fed the capsule were significantly higher than in the control group. These results suggest that incorporation of UL into scallop-derived PL by means of capsulation may lead to an additive increase in the antiobesity properties of these bioactive lipids.

Comparison of 3 spectrophotometric methods for carotenoid determination in frequently consumed fruits and vegetables.

UNLABELLED: Carotenoids are C-40 tetraterpenoid compounds with potential health beneficial effects. Major dietary sources include a variety of fruits and vegetables. Rapid screening methods are therefore desired, but their accuracy varies depending on the carotenoid profile and the matrix of the plant food. In the present study, 3 different methods were compared, all based on a rapid extraction protocol and spectrophotometric measurements to determine the total amount carotenoids present in fruits and vegetables (n = 28), either with or without chlorophyll. Published methods (a) Lichtenthaler and (b) Hornero-Méndez and Mínguez-Mosquera were compared with a newly developed method (method c) based on the average molar absorption coefficient (135310 Lcm(-1)mol(-1)) and wavelength (450 nm in acetone), for the 5 predominant carotenoid species (beta-carotene, zeaxanthin, lycopene, lutein, beta-cryptoxanthin) in the investigated foods. All results were compared to HPLC (method d). To avoid overestimating carotenoid concentrations due to chlorophyll A and B presence, the effect of saponification was studied for all methods. Overall, saponification led to significant carotenoid losses (12.6 +/- 0.9%). Methods a, b, c, and d yielded 5.1 +/- 0.4 mg/100 g, 4.6 +/- 0.5 mg/100 g, 4.3 +/- 0.5 mg/100 g, and 4.2 +/- 0.5 mg/100 g total carotenoids, respectively, with method a leading to significant higher mean concentrations compared to all other methods (P < 0.001, Bonferroni) with methods b and c being not significantly different and highly correlated compared to HPLC (> r = 0.95). Similar results were found when stratifying for chlorophyll content and fruits compared with vegetables, however, accuracy varied for individual fruits, highlighting the limitation to use the same method for all plant foods. PRACTICAL APPLICATION: This study presents a comparison of various rapid spectrophotometric measurements to determine total carotenoid content in various fruits and vegetables and could aid in the selection of the appropriate method for individual plant foods with different carotenoid profile and matrices.

[The assessment and characteristic of dietary supplements with lutein and zeaxanthin on the Polish pharmaceutical market]. / Ocena oraz charakterystyka suplementów diety zawierajacych luteine i zeaksantyne obecnych na polskim rynku farmaceutycznym.

Lutein and zeaxanthin, two xanthophylls supposed to delay formation eye's disease (AMD, cataract), and found in numerous new dietary supplements appearing on market also in Poland. In the observed period on polish pharmaceutical market was evaluated presence about 50 supplements with lutein and 12 contained zeaxanthin as well. The lutein concentration ranges from 0.25 to 20.0 mg/serving size and zeaxanthin from 0.02 to 0.50 mg. Most of them were multicomponent (multi-vitamin products) in form of capsules or tabs for adults, advised for the eyes' protections or supply of nutritional components with various contents of lutein and zeaxanthin and various costs of daily treatment.

Safety assessment of astaxanthin-rich microalgae biomass: Acute and subchronic toxicity studies in rats.

Astaxanthin, a natural nutritional component, is marketed as a dietary supplement around the world. The primary commercial source for astaxanthin is Haematococcus pluvialis (microalgae). The objective of the present study was to investigate the acute and subchronic toxicity of an astaxanthin-rich biomass of H. pluvialis (AstaCarox). The oral LD(50) of the biomass in rats was greater than 12g/kg body weight. In the subchronic study, Wistar rats (10/sex/group) were fed diets containing 0%, 1%, 5% and 20% of the biomass (weight/weight) for 90 days. trans-Astaxanthin was quantifiable in the plasma of the high-dose treated group only. Compared to the control group, no treatment-related biologically significant effects of astaxanthin were noted on body weight or body weight gain. Biomass feeding did not affect hematological parameters. In the high-dose group, slightly elevated alkaline phosphatase and changes in some urine parameters and an increase in kidney weight in both sexes were noted. Histopathology examinations did not reveal adverse effects except for a marginal increase in pigment in the straight proximal tubule of the kidney in 5/10 female rats treated with the high-dose. These changes were not considered as toxicologically significant. Although the rats in high-dose group received about 9% more fat, it is unlikely that this confounding factor significantly altered the outcome. The no-observed adverse-effect-levels (NOAEL) of the astaxanthin-rich biomass for male and female rats were determined as 14,161 and 17,076mg/kg body weight/day, or 465 and 557mg astaxanthin/kg/day, respectively, the highest dose tested.