Insights on the Bioaccessibility of Natural Pigments from Diatom Chaetoceros calcitrans.

This study aimed to investigate the bioaccessibility of carotenoids and chlorophylls from the biomass of microalgae Chaetoceros calcitrans. The samples were submitted to an in vitro digestion protocol, and the compounds were determined by HPLC-PDA-MS/MS. A total of 13 compounds were identified in all tests. After in vitro digestion, the relative bioaccessibility of carotenoids and chlorophylls ranged from 4 to 58%. The qualitative profile of carotenoids reflected the initial sample, with all-E-zeaxanthin (57.2%) being the most bioaccessible compound, followed by all-E-neochrome (31.26%), the latter being reported for the first time in the micellar fraction. On the other hand, among the chlorophylls only pheophytin a (15.01%) was bioaccessible. Furthermore, a chlorophyll derivative (Hydroxypheophytin a') was formed after in vitro digestion. Considering all compounds, xanthophylls (12.03%) and chlorophylls (12.22%) were significantly (p < 0.05) more bioaccessible than carotenes (11.22%). Finally, the considerable individual bioaccessibilities found, especially for zeaxanthin, demonstrate the bioactive potential of this bioresource. However, the large reduction in the totality of compounds after in vitro digestion suggests that additional technological strategies should be explored in the future to increase the efficiency of micellarization and enhance its bioactive effects.

Alternative green solvents associated with ultrasound-assisted extraction: A green chemistry approach for the extraction of carotenoids and chlorophylls from microalgae.

This study proposes a method for the ultrasonic extraction of carotenoids and chlorophyll from Scenedesmus obliquus and Arthrospira platensis microalgae with green solvents. Ethanol and ethanolic solutions of ionic liquids were tested with a variety of extraction parameters, including number of extractions, time of extraction, and solid-liquid ratio R(S/L), to determine the optimal conditions. After selecting the most effective green solvent (ethanol), the process conditions were established: R(S/L) of 1:10, three extraction cycles at 3 min each), giving an extraction yield of 2602.36 and 764.21 µgcarotenoids.gdried biomass-1; and 22.01 and 5.81 mgchlorophyll.gdried biomass-1 in S. obliquus and A. platensis, respectively. The carotenoid and chlorophyll extracts obtained using ethanol were shown to be potent scavengers of peroxyl radical, being 5.94 to 26.08 times more potent α-tocopherol. These findings pave the way for a green strategy for valorizing microalgal biocompounds through efficient and environmentally friendly technological processes.

Guidance for formulating ingredients/products from Chlorella vulgaris and Arthrospira platensis considering carotenoid and chlorophyll bioaccessibility and cellular uptake.

This study aimed to investigate the impact of different microalgal matrices on the bioaccessibility and uptake by Caco-2 cells of carotenoids and chlorophylls. In this way, the microalgal ingredients/products (whole dry biomass [WDB], whole ultrasonicated paste [WUP], and liposoluble pigment emulsion [LPE]) obtained from Chlorella vulgaris and Arthrospira platensis were submitted to in vitro simulated digestion. Apical uptake of pigments in micelles generated during the simulated digestion by Caco-2 human intestinal cells was determined. The influence of simulated digestion on carotenoid and chlorophyll stability and bioaccessibility was assessed by HPLC-PDA-MS/MS and the carotenoids and chlorophylls' bioaccessibility and cellular uptake were shown to be boosted according to the matrix (LPE > WUP > WDB). Our findings showed that Chlorella vulgaris and Arthrospira platensis could be considered in formulations when carotenoids and chlorophylls are the target molecules in the ingredients/products.

Bioaccessibility of microalgae-based carotenoids and their association with the lipid matrix.

The composition of microalgae can contribute to nutritious and functional diets. Among the functional compounds, carotenoids are in focus since positive effects on human health have been established, which are in turn related to their bioaccessibility. In addition to essential nutrients, our hypothesis was that microalgae biomasses could be used as sources of bioaccessible carotenoids. Thus, this study determined for the first time the bioaccessibility of carotenoids from biomass of Scenedesmus bijuga and Chlorella sorokiniana and their possible relationship with the lipid composition of the matrix. The samples were submitted to in vitro digestion protocol, and carotenoids were determined by HPLC-PDA-MS/MS. Individual bioaccessibility of carotenoids was ≥ 3.25%. In general, compounds in their cis conformation were more bioaccessible than trans; and total carotenes more than total xanthophylls. Twelve compounds were bioaccessible from the biomass of S. bijuga, and eight in C. sorokiniana. In S. bijuga, the bioaccessibility of total carotenoids was 7.30%, and the major bioaccessible carotenoids were 9-cis-ß-carotene (43.78%), 9-cis-zeaxanthin (42.30%) followed by 9-cis-lutein (26.73%); while in C. sorokiniana, the total bioaccessibility was 8.03%, and 9-cis-ß-carotene (26.18%), all-trans-ß-carotene (13.56%), followed by 13-cis-lutein (10.71%) were the major compounds. Overall, the total content of lipids does not influence the bioaccessibility of total carotenoids. Still, the lipid composition, including structural characteristics such as degree of saturation and chain length of the fatty acid, impacts the promotion of individual bioaccessibility of carotenes and xanthophylls of microalgae. Finally, the results of this study can assist the development of microalgae-based functional food ingredients and products.

Insights on the intestinal absorption of chlorophyll series from microalgae.

The bioaccessibility and subsequent uptake by Caco-2 human intestinal cells of chlorophyll pigments from Scenedesmus obliquus were determined for the first time. In order to evaluate the impact of different types of the matrix on bioaccessibility of chlorophyll from microalgae, three different products were evaluated: isolated chlorophyll extract (ICE); wet ultrasonicated biomass (WUB); and whole dried biomass (WDB). The samples were submitted to in vitro digestion model according to the INFOGEST protocol, and Caco-2 cells determined the intestinal uptake. Chlorophyll pigments were determined by HPLC-PDA-MS/MS. A total of ten chlorophyll pigments (8,318.48 µg g-1) were separated in S. obliquus biomass, with chlorophyll a (3,507.76 µg g-1) and pheophytin a' (1,598.09 µg g-1) the major ones. After in vitro digestion, all tested products showed bioaccessible chlorophylls. However, the total bioaccessibility results were as follows: ICE (33.45%), WUB (2.65%), WDB (0.33%). Five compounds were bioaccessible in ICE, three in WUB, and one in WDB. The hydroxypheophytin a showed the highest bioaccessibility (212%) in ICE, while pheophytin a' in WUB (11%) and WDB (2%). As a result, bioavailability estimates of ICE using the Caco-2 cell showed hydroxypheophytin a (102.53%), followed by pheophytin a' (64.69%) as the chlorophyll pigments most abundant in intestinal cells. In summary, from a nutritional perspective, these three types of the matrix (WDB, WUB, and ICE) influence the promotion of chlorophyll bioaccessibility. In this way, the data suggest that chlorophylls bioaccessibility from ICE is greater than that in WDB and WUB. Therefore, ICE should be considered a product that provides bioavailable chlorophyll and could be the best choice, such as ingredients in the development of functional foods chlorophyll-based.

HPLC-PDA-MS/MS as a strategy to characterize and quantify natural pigments from microalgae.

Interest in pigment composition of microalgae species is growing as new natural pigments sources are being sought. However, we still have a limited number of species of microalgae exploited to obtain these compounds. Considering these facts, the detailed composition of carotenoids and chlorophylls of two species of green microalgae (Chlorella sorokiniana and Scenedesmus bijuga) were determined for the first time by high-performance liquid chromatography coupled to diode array and mass spectrometry detectors (HPLC-PDA-MS/MS). A total of 17 different carotenoids were separated in all the extracts. Most of the carotenoids present in the two microalgae species are xanthophylls. C. sorokiniana presented 11 carotenoids (1408.46 µg g-1), and S. bijuga showed 16 carotenoids (1195.75 µg g-1). The main carotenoids detected in the two microalgae were all-trans-lutein and all-trans-ß-carotene. All-trans-lutein was substantially higher in C. sorokiniana (59.01%), whereas all-trans-ß-carotene was detected in higher quantitative values in S. bijuga (13.88%). Seven chlorophyll compounds were identified in both strains with different proportions in each species. Concentrations of chlorophyll representing 7.6% and 10.2% of the composition of the compounds present in the biomass of C. sorokiniana and S. bijuga, respectively. Relevant chlorophyll compounds are reported for the first time in these strains. The data obtained provide significant insights for microalgae pigment composition databases.

Exploratory data of the microalgae compounds for food purposes.

This brief data article refers to the previous exploration of Scenedesmus obliquus and Phormidium autumnale biomass about the possibility of using these microalgae species as an unconventional functional food. Data on chemical composition, fatty acids, volatile compounds, and carotenoid profiles were determined. In parallel, are provided the antioxidant capacity (reducing capacity - RC and reactive oxygen species deactivation - ORAC) of aqueous, lipophilic, and carotenoid extracts isolated from microalgae biomass. Both species have similar compounds in their biomass. However, S. obliquus was statistically different with a lower saturated fatty acid (STF) followed by higher mono (MUFA) and polyunsaturated (PUFA) content, also showed higher antioxidant potential for acetone extract and isolated carotenoids. On the other hand, P. autumnale aqueous extract showed high RC and ORAC. The significance of the experimental data was determined using the t-test (p < 0.05) based on the Statistica 7.0 software. These findings led us to explore the microalgae S. obliquus in an in vivo experimental model.

Carotenoid profile of three microalgae/cyanobacteria species with peroxyl radical scavenger capacity.

Carotenoids from cyanobacteria Aphanothece microscopica Nageli and green microalgae Chlorella vulgaris and Scenedesmus obliquus were identified. The total carotenoid content, based on dry weight of biomass, of A. microscopica Nägeli, C. vulgaris and S. obliquus were 1398.88µg/g, 1977.02µg/g and 2650.70µg/g, respectively. A total of 23 different carotenoids were separated in all the extracts, the major ones being all-trans-ß-carotene (29.3%) and all-trans-lutein (28.1%) in Scenedesmus; all-trans-echinenone (22.8%) and all-trans-ß-carotene (17.7%) in Chlorella; all-trans-echinenone (28.3%) and all-trans-ß-carotene (26.2%) in Aphanothece. The carotenoid extracts were shown to be a potent scavenger of peroxyl radical, with values of 31.1 (Chlorella), 14.0 (Scenedesmus) and 7.3 (Aphanothece) times more potent than α-tocopherol.

Identification of chlorophyll molecules with peroxyl radical scavenger capacity in microalgae Phormidium autumnale using ultrasound-assisted extraction.

The identification of chlorophyll molecules with peroxyl radical scavenger capacity in microalgae Phormidium autumnale was determined. The ultrasound-assisted extraction was utilized for obtaining the chlorophyll compounds from biomass. A total of eleven molecules were separated in microalgae chlorophyll extract, with pheophytin a' (371µg·g-1) and chlorophyll a (159.3µg·g-1) as the major ones. The chlorophyll extract was shown to be a potent scavenger of peroxyl radical, being almost 200 times more potent than α-tocopherol. These facts suggest the microalgae Phormidium autumnale as potential source of bioactive tetrapyrrole compounds.