Ladislava Mišurcová

Bioactive phenols in algae: The application of pressurized-liquid and solid-phase extraction techniques

A new extraction technique based on the off-line combination of pressurized-liquid with solid-phase extraction (PLE-SPE) is described. The method was used for the extraction of bioactive phenolic acids (protocatechuic, p-hydroxybenzoic, 2,3-dihydroxybenzoic, chlorogenic, vanillic, caffeic, p-coumaric, salicylic acid), cinnamic acid and hydroxybenzaldehydes (p-hydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde, vanillin) from in vitro culture of two freshwater algae (Anabaena doliolum and Spongiochloris spongiosa) and from food products of marine macroalgae Porphyra tenera (nori) and Undaria pinnatifida (wakame). For the identification and quantification of the compounds the molecular ions [M-H](-) and specific fragments were analyzed by quadrupole mass spectrometry analyzer connected on-line with a reversed-phase HPLC system. Our analysis showed that the freshwater algae and marine algal products contained submicrogram or microgram level of above-mentioned phenols per gram of lyophilized sample. In addition, the total phenol content (Folin-Ciocalteu assay) and antioxidant activity (TEAC assay, Trolox equivalent antioxidant capacity assay) of the PLE-SPE extracts were determined and discussed.

Fatty Acids Composition of Vegetable Oils and Its Contribution to Dietary Energy Intake and Dependence of Cardiovascular Mortality on Dietary Intake of Fatty Acids

Characterizations of fatty acids composition in % of total methylester of fatty acids (FAMEs) of fourteen vegetable oils—safflower, grape, silybum marianum, hemp, sunflower, wheat germ, pumpkin seed, sesame, rice bran, almond, rapeseed, peanut, olive, and coconut oil—were obtained by using gas chromatography (GC). Saturated (SFA), monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA), palmitic acid (C16:0; 4.6%–20.0%), oleic acid (C18:1; 6.2%–71.1%) and linoleic acid (C18:2; 1.6%–79%), respectively, were found predominant. The nutritional aspect of analyzed oils was evaluated by determination of the energy contribution of SFAs (19.4%–695.7% ERDI), PUFAs (10.6%–786.8% ERDI), n-3 FAs (4.4%–117.1% ERDI) and n-6 FAs (1.8%–959.2% ERDI), expressed in % ERDI of 1 g oil to energy recommended dietary intakes (ERDI) for total fat (ERDI—37.7 kJ/g). The significant relationship between the reported data of total fat, SFAs, MUFAs and PUFAs intakes (% ERDI) for adults and mortality caused by coronary heart diseases (CHD) and cardiovascular diseases (CVD) in twelve countries has not been confirmed by Spearman’s correlations.

Antioxidant Activity and Protecting Health Effects of Common Medicinal Plants

Medicinal plants are traditionally used in folk medicine as natural healing remedies with therapeutic effects such as prevention of cardiovascular diseases, inflammation disorders, or reducing the risk of cancer. In addition, pharmacological industry utilizes medicinal plants due to the presence of active chemical substances as agents for drug synthesis. They are valuable also for food and cosmetic industry as additives, due to their preservative effects because of the presence of antioxidants and antimicrobial constituents. To commonly used medicinal plants with antioxidant activity known worldwide belong plants from several families, especially Lamiaceae (rosemary, sage, oregano, marjoram, basil, thyme, mints, balm), Apiaceae (cumin, fennel, caraway), and Zingiberaceae (turmeric, ginger). The antioxidant properties of medicinal plants depend on the plant, its variety, environmental conditions, climatic and seasonal variations, geographical regions of growth, degree of ripeness, growing practices, and many other factors such as postharvest treatment and processing. In addition, composition and concentration of present antioxidants, such as phenolic compounds, are related to antioxidant effect. For appropriate determination of antioxidant capacity, the extraction technique, its conditions, solvent used, and particular assay methodology are important.

Health benefits of algal polysaccharides in human nutrition.

The interest in functional food, both freshwater and marine algal products with their possible promotional health effects, increases also in regions where algae are considered as rather exotic food. Increased attention about algae as an abundant source of many nutrients and dietary fiber from the nutrition point of view, as well as from the scientific approaches to explore new nutraceuticals and pharmaceuticals, is based on the presence of many bioactive compounds including polysaccharides extracted from algal matter. Diverse chemical composition of dietary fiber polysaccharides is responsible for their different physicochemical properties, such as their ability to be fermented by the human colonic microbiota resulted in health benefit effects. Fundamental seaweed polysaccharides are presented by alginates, agars, carrageenans, ulvanes, and fucoidans, which are widely used in the food and pharmaceutical industry and also in other branches of industry. Moreover, freshwater algae and seaweed polysaccharides have emerged as an important source of bioactive natural compounds which are responsible for their possible physiological effects. Especially, sulfate polysaccharides exhibit immunomodulatory, antitumor, antithrombotic, anticoagulant, anti-mutagenic, anti-inflammatory, antimicrobial, and antiviral activities including anti-HIV infection, herpes, and hepatitis viruses. Generally, biological activity of sulfate polysaccharides is related to their different composition and mainly to the extent of the sulfation of their molecules. Significant attention has been recently focused on the use of both freshwater algae and seaweed for developing functional food by reason of a great variety of nutrients that are essential for human health.

Phenolic content and antioxidant capacity in algal food products

The study objective was to investigate total phenolic content using Folin-Ciocalteu’s method, to assess nine phenols by HPLC, to determine antioxidant capacity of the water soluble compounds (ACW) by a photochemiluminescence method, and to calculate the correlation coefficients in commercial algal food products from brown (Laminaria japonica, Eisenia bicyclis, Hizikia fusiformis, Undaria pinnatifida) and red (Porphyra tenera, Palmaria palmata) seaweed, green freshwater algae (Chlorella pyrenoidosa), and cyanobacteria (Spirulina platensis). HPLC analysis showed that the most abundant phenolic compound was epicatechin. From spectrophotometry and ACW determination it was evident that brown seaweed Eisenia bicyclis was the sample with the highest phenolic and ACW values (193 mg·g−1 GAE; 7.53 µmol AA·g−1, respectively). A linear relationship existed between ACW and phenolic contents (r = 0.99). Some algal products seem to be promising functional foods rich in polyphenols.

Seaweed Minerals as Nutraceuticals

Seaweed is known as an abundant source of minerals. Mineral composition of seaweed is very changeable because of many exogenous and endogenous factors and differs also within the same species. Principally, seaweed is an excellent source of some essential elements. Mainly, iron and iodine are in high concentration. Seaweeds could be prospective as functional foods and also producers of mineral nutraceuticals.

Seaweed lipids as nutraceuticals.

Seaweeds are known as low-energy food. Despite low lipid content, ω-3 and ω-6 polyunsaturated fatty acids (PUFAs) introduce a significant part of seaweed lipids. PUFAs are the important components of all cell membranes and precursors of eicosanoids that are essential bioregulators of many cellular processes. PUFAs effectively reduce the risk of cardiovascular diseases, cancer, ostheoporosis, and diabetes. Because of the frequent usage of seaweeds in Asia and their increasing utilization as food also in other parts of the world, seaweeds could contribute to the improvement of a low level of ω-3 PUFAs, especially in the Western diet. The major commercial sources of ω-3 PUFAs are fish, but their wide usage as food additives is limited for the typical fishy smell, unpleasant taste, and oxidative nonstability. Nevertheless, growing requirements of healthy functional foods have led to produce PUFAs as nutraceuticals in controlled batch culture of marine microalgae, especially Thraustochytrium and Schizochytrium strains.

Amino acid composition of algal products and its contribution to RDI

In this paper, the amino acid profiles of algal products from diverse groups (Cyanophyceae, Chlorophyta, Rhodophyta and Phaeophyta) were established. Contents of essential and non-essential amino acids varied in the range of 22.8-42.3 and 31.0-66.5 (g·16 g(-1)N), respectively. In dependence on daily algal intakes, the highest participation in recommended daily intakes (RDI; related to adult, body weight of 70 kg) of all EAAs was observed in freshwater micro-algal products, especially from Spirulina genus where contribution ranged from 12.6% (Lys, SB) to 38.8% (Thr, S). Generally, Lys was the lowest contributor to RDIs in almost all algal samples except Chlorella pyrenoidosa (C) and Palmaria palmata (D), where Ile and Leu, respectively, were established. Interestingly, the contents of sulfur AAs of both products of the Spirulina genus covered 74.5% (S) and 73.8% (SB) of their RDI. Finally, products from brown seaweeds showed the lowest contributions to the RDIs of all EAAs.

Influence of Extractive Solvents on Lipid and Fatty Acids Content of Edible Freshwater Algal and Seaweed Products, the Green Microalga Chlorella kessleri and the Cyanobacterium Spirulina platensis

Total lipid contents of green (Chlorella pyrenoidosa, C), red (Porphyra tenera, N; Palmaria palmata, D), and brown (Laminaria japonica, K; Eisenia bicyclis, A; Undaria pinnatifida, W, WI; Hizikia fusiformis, H) commercial edible algal and cyanobacterial (Spirulina platensis, S) products, and autotrophically cultivated samples of the green microalga Chlorella kessleri (CK) and the cyanobacterium Spirulina platensis (SP) were determined using a solvent mixture of methanol/chloroform/water (1:2:1, v/v/v, solvent I) and n-hexane (solvent II). Total lipid contents ranged from 0.64% (II) to 18.02% (I) by dry weight and the highest total lipid content was observed in the autotrophically cultivated cyanobacterium Spirulina platensis. Solvent mixture I was found to be more effective than solvent II. Fatty acids were determined by gas chromatography of their methyl esters (% of total FAMEs). Generally, the predominant fatty acids (all results for extractions with solvent mixture I) were saturated palmitic acid (C16:0; 24.64%–65.49%), monounsaturated oleic acid (C18:1(n-9); 2.79%–26.45%), polyunsaturated linoleic acid (C18:2(n-6); 0.71%–36.38%), α-linolenic acid (C18:3(n-3); 0.00%–21.29%), γ-linolenic acid (C18:3(n-6); 1.94%–17.36%), and arachidonic acid (C20:4(n-6); 0.00%–15.37%). The highest content of ω-3 fatty acids (21.29%) was determined in Chlorella pyrenoidosa using solvent I, while conversely, the highest content of ω-6 fatty acids (41.42%) was observed in Chlorella kessleri using the same solvent.

In Vitro Digestibility of Different Commercial Edible Algae Products

Digestibility is an important factor in determining the level of nutritive factors utilization. Determination of in vitro digestibility in a Daisy incubator and assessment of protein digestibility using Kjeldahl method of algal food products after pepsin, pancreatin, and combined hydrolysis of both enzymes was described. The influence of different hydrolysis times on algal digestibility was investigated in brown seaweeds, red seaweeds, and green alga. The highest digestibility values were generally determined in red seaweeds after 24 h of combined hydrolysis by pepsin and pancreatin. Direct dependence between digestion time and digestion efficiency was verified and statistically proven.