Influence of long term nitrogen limitation on lipid, protein and pigment production of Euglena gracilis in photoheterotrophic cultures.

Nitrogen limitation is considered a good strategy for enhancement of algal lipid production while conversely N repletion has been shown to result in biomass rich in proteins. In this study, the influence of long-term N limitation on Euglena gracilis fatty acid (FA), protein, chlorophyll a, and carotenoid concentrations was studied in N limited cultures. Biomass composition was analyzed from three-time points from N starved late stationary phase cultures, exposed to three different initial N concentrations in the growth medium. Total lipid content increased under N limitation in ageing cultures, but the low N content and prolonged cultivation time resulted in the formation of a high proportion of saturated FAs. Furthermore, growth as well as the production of proteins, chlorophyll a and carotenoids were enhanced in higher N concentrations and metabolism of these cellular components stayed stable during the stationary growth phase. Our findings showed that a higher N availability and a shorter cultivation time is a good strategy for efficient E. gracilis biomass production, regardless of whether the produced biomass is intended for maximal recovery of polyunsaturated FAs, proteins, or photosynthetic pigments. Additionally, we showed an increase of neoxanthin, ß-carotene, and diadinoxanthin as a response to higher N availability.

The lager yeast Saccharomyces pastorianus removes and transforms Fusarium trichothecene mycotoxins during fermentation of brewer's wort.

An investigation was conducted to determine the fate of deoxynivalenol, deoxynivalenol-3-glucoside, HT-2 toxin and T-2 toxin, during a four-day fermentation with the lager yeast Saccharomyces pastorianus. The influence of excessive mycotoxin concentrations on yeast growth, productivity and viability were also assessed. Mycotoxins were dosed at varying concentrations to 11.5° Plato wort. Analysis of yeast revealed that presence of the toxins even at concentrations up to 10,000 µg/L had little or no effect on sugar utilisation, alcohol production, pH, yeast growth or cell viability. Of the dosed toxin amounts 9-34% were removed by the end of fermentation, due to physical binding and/or biotransformation by yeast. Deoxynivalenol-3-glucoside was not reverted to its toxic precursor during fermentation. Processing of full-scan liquid chromatography-quadrupole time-of-flight-mass spectrometry (LC-QTOF-MS) data with MetaboLynx and subsequent LC-QTOF-MS/MS measurements resulted in annotation of several putative metabolites. De(acetylation), glucosylation and sulfonation were the main metabolic pathways activated.

EuroFIR guidelines for assessment of methods of analysis: GAMA.

Analytical methodology is a key factor in food composition databases and specific criteria, at the component level, is needed for comparison of analytical data from different sources. The aim of this work is to describe how EuroFIR guidelines for assessment of methods of analysis are created and made available to users. Comprehensive information for macronutrients, vitamins, minerals and trace elements addressing all aspects of analytical procedures was obtained from international standards, and scientific literature. Documentation was compiled in a confluence wiki format provided for each component: background information, description of reference methods of analysis and critical steps, available reference materials, proficiency testing schemes, other analytical methods and relevant references. The information for each nutrient was collated, edited and presented with hypertext links to additional pages where more detailed information can be accessed using full text searches. The wiki format is a useful tool for preparing information and disseminating to users.

Ultra-high performance liquid chromatographic and mass spectrometric analysis of active vitamin B12 in cells of Propionibacterium and fermented cereal matrices.

A sensitive and selective method is needed to analyse in situ produced vitamin B12 in plant-based materials, potential new dietary sources of vitamin B12. A UHPLC/UV method was developed and validated for the determination of human active vitamin B12 in cell extracts of Propionibacterium freudenreichii subsp. shermanii and after immunoaffinity purification in extracts of cereal matrices fermented by P. freudenreichii. An Acquity HSS T3 C18 column resulted in a baseline separation, a calibration curve of excellent linearity and a low limit of detection (0.075 ng/5 µL injection). As confirmed by UHPLC-MS, the active vitamin B12 could be separated from pseudovitamin B12. The recovery of vitamin B12 from purified spiked cereal matrices was good (>90%; RSD<5%). A nutritionally relevant amount of active vitamin B12 was produced by P. freudenreichii in cereal malt matrices (up to 1.9 µg/100 g) in 24h at 28 °C.

Antioxidant activity of isolated ellagitannins from red raspberries and cloudberries.

Ellagitannins from red raspberries (Rubus idaeus) and cloudberries (Rubus chamaemorus) were isolated by using column chromatography and preparative HPLC. The berry phenolic isolates consisted of 80% (cloudberry) and of 60% (raspberry) of ellagitannins, with raspberries also containing anthocyanins. The main ellagitannins of both raspberries and cloudberries were identified by ESI-MS to consist of the dimeric sanguiin H-6 and the trimeric lambertianin C. Monomeric ellagitannins such as casuarictin in raspberries and pedunculagin in cloudberries were also found. The antioxidant activity of the berry phenolic isolate, ellagitannin isolate (mixture), ellagitannin main fraction (dimer and trimer), and ellagic acid was studied in bulk and emulsified methyl linoleate, in human low-density lipoprotein in vitro, and the radical scavenging activity was studied in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test. Cloudberry and red raspberry ellagitannins were highly effective as radical scavengers. Berry ellagitannins also showed significant antioxidant activity toward oxidation of both human LDL and methyl linoleate emulsions. However, only weak or moderate antioxidant activity was exhibited by ellagitannins toward oxidation of bulk oil. Thus, ellagitannins contribute significantly to the antioxidant capacity of cloudberries and red raspberries in lipoprotein and lipid emulsion environments, the latter being more relevant for food applications.

Separation and identification of neutral cereal lipids by normal phase high-performance liquid chromatography, using evaporative light-scattering and electrospray mass spectrometry for detection.

A novel method was developed for the analysis of molecular species in neutral lipid classes, using separation by normal phase high-performance liquid chromatography, followed by detection by evaporative light-scattering and electrospray ionization tandem mass spectrometry. Monoacid standards, i.e. sterol esters, triacylglycerols, fatty acids, diacylglycerols, free sterols and monoacylglycerols, were separated to baseline on microbore 3 microm-silica gel columns. Complete or partial separation of molecular species in each lipid class permitted identification by automatic tandem mass spectrometry of ammonium adducts, produced via positive electrospray ionization. After optimization of the method, separation and identification of molecular species of various lipid classes was comprehensively tested by analysis of neutral lipids from the free lipid extract of maize flour.

Analysis of protein oxidation markers alpha-aminoadipic and gamma-glutamic semialdehydes in food proteins using liquid chromatography (LC)-electrospray ionization (ESI)-multistage tandem mass spectrometry (MS).

To elucidate the formation of protein oxidation biomarkers alpha-aminoadipic semialdehyde (AAS) and gamma-glutamic semialdehyde (GGS) in food proteins was the main purpose of the present study. Food proteins, namely, myofibrillar proteins, alpha-lactalbumin, and soy proteins, as well as bovine serum albumin (BSA), were suspended in a piperazine-1,4-bis(2-ethanesulfonic acid) (PIPES) buffer and oxidized by Fe(3+) and H(2)O(2) while kept in an oven for 14 days at 37 degrees C. For the analysis of semialdehydes, a derivatization procedure with p-aminobenzoic acid (ABA) and NaCNBH(3) followed by liquid chromatography (LC)-electrospray ionization (ESI)-multistage tandem mass spectrometry (MS) was performed. For comparative purposes, the dinitrophenylhydrazine (DNPH) method was also employed as a routine method to assess carbonyl gain. Both semialdehydes were specifically and accurately detected by LC-MS in all oxidized proteins proving that GGS and AAS are formed as a consequence of the oxidation of lysine, proline, and arginine amino acid residues from BSA and other food proteins. Proteins from an animal source and, particularly, BSA were more susceptible to undergo oxidative reactions than soy proteins. The results from the present paper highlight the significance of using both semialdehydes as protein oxidation indicators in meat and dairy products. The analysis of GGS and AAS in real food systems would contribute to the understanding of the precise mechanisms involved in food protein oxidation and shed light on the fate of oxidizing amino acids during food processing and storage.

Acetylcholinesterase inhibitory guided fractionation of Melissa officinalis L.

The plant Melissa officinalis L. has been used traditionally in the treatment of cognitive dysfunction. Based on its traditional medicinal use, it was assessed for its clinical efficacy in mild to moderate Alzheimer's patients. The plant was effective in the management of the disease. Therefore, based on this result, a similar plant extract was prepared in order to be screened for bioactivities which are relevant in Alzheimer's disease therapy. The extract was recently screened for antioxidant activity and it showed a wide range of antioxidant properties. Another important bioactivity is acetylcholinesterase inhibition, which the extract was screened for in the current investigation. The extract was capable of inhibiting the enzyme in a time and dose-dependent manner. Activity of the extract at 10 min was estimated as 1.72+/-0.16 microg equivalents of physostigmine/mg of the extract. Acetylcholinesterase inhibitory guided fractionation of the extract was then carried out. Most of the fractions showed inhibitory activity and were more potent than the extract. The contents of the most potent fraction were identified as cis- and trans-rosmarinic acid isomers and a rosmarinic acid derivative using LC-DAD-ESI-MS and NMR methods.

Tocopherols and tocotrienols in wheat genotypes in the HEALTHGRAIN Diversity Screen.

Tocopherol and tocotrienol compositions were studied in 175 genotypes of different wheat types grown under similar conditions to screen for natural diversity. The main focus was on bread wheats, including 130 and 20 winter and spring types, respectively. The average total content of tocopherols and tocotrienols was 49.4 microg/g of dm, with a range of 27.6-79.7 microg/g of dm, indicating a 2.9-fold variation among genotypes. Beta-tocotrienol and alpha-tocopherol were the major vitamers, and in general there were more tocotrienols than tocopherols. In the early cultivated forms of wheat the proportion of tocotrienols was especially high, at >or=62.5%. In conclusion, there was a large variation in total tocopherol and tocotrienol contents in bread wheats and this, along with the high proportions of tocotrienols in other types of wheat, demonstrates the great genetic potential of genotypes to be exploited by plant breeders.

Potential of selected infant food formulas for production of Bacillus cereus emetic toxin, cereulide.

Cereulide producing Bacillus cereus was isolated from randomly chosen commercial infant foods. The cereulide production in infant food formulas was investigated. When the reconstituted foods were inoculated with >10(5) cfu ml(-1) of cereulide producing B. cereus, 2 to 200 microg of cereulide per 100 ml of food accumulated during 24 h of non-refrigerated storage. The amount of cereulide measured in the foods by the accurate chemical assay (LC-MS) matched with that found by sperm micro assay, proving the cereulide was the sole heat stable toxin in the foods and present in its toxic form. The infant formulas containing both cereal and dairy ingredients were the most supportive for cereulide production. Cereulide accumulation was affected by the infant food composition as well as by the handling of the food. Diluting the reconstituted food with water resulted in increased toxin production expressed as mug per volume. More cereulide was accumulated when the food was incubated stationary compared with moderate shaking. The amount of cereulide accumulated within 24 h at room temperature per 100 ml of cereal and dairy or in rice-nondairy reconstituted infant formulas, inoculated with >or=10(5) cfu ml(-1) of B. cereus strain F4810/72, was higher or similar to the amounts reported for foods implicated in emetic type of food poisonings. Thus mishandling and temperature abuse of infant foods may cause food poisoning when emetic B. cereus is present.

Influence of incorporated wild Solanum genomes on potato properties in terms of starch nanostructure and glycoalkaloid content.

Interspecific somatic hybrids produced by protoplast fusion between two wild Solanum species (S. acaule, acl; S. brevidens, brd) and cultivated potato Solanum tuberosum (tbr) were analyzed in terms of the starch nanometer-range structure and glycoalkaloid (GA) contents. The crystallinity of starch granules, the average size of starch crystallites, and the lamellar distances were obtained from tuber samples using wide-angle and small-angle X-ray scattering methods. These measurements showed that incorporation of wild genomes from either nontuberous (brd) or tuberous (acl) Solanum species caused no significant modifications of the nanostructure of potato starch. In contrast, the GA profiles of the hybrids, which were analyzed by LC-ESI-MS in both tuber and foliage samples, differed considerably from those of cultivated potato. Regardless of the low total tuber GA concentrations (approximately 9 mg/100 g of fresh weight), the somatic hybrids contained GAs not detected in the parental species. A high proportion of spirotype GAs consisting of 5,6-dihydrogenated aglycons, for example, alpha-tomatine and tomatidine bound with solatriose, and chacotriose were found in the hybrids. In conclusion, the foliage of interspecific hybrids contained a higher variation in the structures of GAs than did the tubers.

Effect of selenate supplementation on glycoalkaloid content of potato (Solanum tuberosum L.).

Potatoes (Solanum tuberosum L.) supplemented with increasing amounts of sodium selenate were analyzed for glycoalkaloid (GA) content. GAs were extracted with 5% acetic acid from freeze-dried tubers of two potato cultivars, Satu and Sini, harvested 10 weeks after planting as immature. The GAs alpha-solanine and alpha-chaconine were quantified by reverse-phase high-performance liquid chromatography (RP-HPLC) with diode array detection. Two independent experiments were performed. In the first experiment, the total GA concentration +/- standard error of the tubers ranged between 105 +/- 9 and 124 +/- 10 mg kg(-1) fresh weight in Satu and between 194 +/- 26 and 228 +/- 10 mg kg(-1) fresh weight in Sini. The ratio of alpha-solanine to alpha-chaconine was 0.2 in Satu and 0.5-0.6 in Sini. In the second experiment, the total GA concentration +/- standard error was 75 +/- 4 to 96 +/- 11 mg kg(-1) fresh weight, and the ratio of alpha-solanine to alpha-chaconine was 0.3-0.4 in Satu. A high sodium selenate supplementation (0.9 mg of Se kg(-1) quartz sand) slightly decreased the GA content in Satu, but this decrease was not statistically significant. Furthermore, at this addition level the Se concentration increased to a very high level of 20 microg g(-1) dry weight, which cannot be recommended for human consumption. In both experiments, the Se concentration in tubers increased with increasing sodium selenate application levels. Our results show that acceptable application levels of selenate did not have an effect on the GA concentration in immature potato tubers.

Steryl phenolic acid esters in cereals and their milling fractions.

The steryl ferulate contents of rye and wheat grains and their milling fractions were analyzed using a reversed-phase high-performance liquid chromatographic (HPLC) method. HPLC-mass spectrometry was used for identification. In addition, steryl ferulates of some selected milling byproducts were determined. The total steryl ferulate contents of rye and wheat grains were 6.0 and 6.3 mg/100 g, respectively. Uneven distribution of steryl ferulates in the grains led to considerable differences in the milling products; their steryl ferulate contents ranged from trace amounts in flours with low ash content to 20 and 34 mg/100 g in rye and wheat brans, respectively. Campestanyl ferulate and sitostanyl ferulate were the main components, followed by campesteryl ferulate and sitosteryl ferulate, whereas sitosterol was the main component in total sterols. Among the other samples, a byproduct of rice milling (pearling dust) was the best source of steryl ferulates, its total steryl ferulate content being 119 mg/100 g, whereas no measurable amounts of steryl ferulates were measured in oat bran or pearling dust of barley. The results indicated that rye and wheat and especially their bran fractions are comparable to corn as steryl ferulate sources.