Amino acid composition of wheat grain. 1: Changes during development

The changes in the composition of free and total amino acids in wheat grain during development are described: the experiment was performed in a growth chamber with three varieties of winter wheat (Triticum aestivum L.): Splendeur, Hobbit and Maris Huntsman. The changes in the total composition of amino acids of the grain as development progressed were consistent with an increase in the proportion of storage proteins. At 15 days after anthesis, the relative amount of glycine increased, suggesting that at this moment the proportion of gliadins was relatively low. The relative amount of arginine increased in mature grain, showing that at times close to maturity the relative proportion of synthesis of a protein fraction rich in this amino acid increased. Free glutamate, proline and arginine decreased at 15 days to a greater extent than the other amino acids and was present in low relative amounts during the period of maximum protein synthesis in the grain; these three amino acids could be the most limiting for protein synthesis in grain. In the free fraction the following showed a similar pattern of change during grain development: aspartate, arginine and histidine on the one hand, and on the other alanine, valine, leucine, isoleucine, threonine, phenylalanine and tyrosine; moreover both groups were inversely correlated. Glycine and lysine were directly related to each other and inversely to glutamate.     

HPLC flavonoid profiles as markers for the botanical origin of European unifloral honeys

The HPLC phenolic profiles of 52 selected unifloral honey samples produced in Europe were analysed to detect possible markers for the floral origin of the different honeys. Lime‐tree (five markers), chestnut (five markers), rapeseed (one marker), eucalyptus (six markers) and heather (three markers) honeys had specific markers with characteristic UV spectra. In addition, the flavanone hesperetin was confirmed as a marker for citrus honey, as well as kaempferol for rosemary honey and quercetin for sunflower honey. Abscisic acid, which had been reported to be a possible marker for heather honey, was also detected in rapeseed, lime‐tree and acacia honeys. Ellagic acid in heather honey and the hydroxycinnamates caffeic, p‐coumaric and ferulic acids in chestnut, sunflower, lavender and acacia honeys were also detected. The characteristic propolis‐derived flavonoids pinocembrin, pinobanksin and chrysin were present in most samples in variable amounts. © 2001 Society of Chemical Industry     

HPLC/UV determination of organic acids in fruit juices and nectars

A reversed phase HPLC method for separation and determination of organic acids in fruit juices and nectars is presented. The method is based on the reaction of free organic acids with O-(4 nitrobenzyl)-N,N'-diisopropylisourea (PNBDI) in presence of dioxane. Excess of reagent was removed with a strong cation-exchange resin. The p-nitrobenzyl esters were separated on a C₁₈ reversed phase column using gradient elution with water and acetonitrile and UV detection at 265 nm. Benzylmalonic acid was used as internal standard. The determinations were performed in the linear range of 0.05-2 g/l for lactic, acetic and succinic acids, 0.1-12 g/l for tartaric, malic and citric acids. The detection limits were 0.025 g/l, 0.017 g/l, 0.050 g/l, 0.039 g/l, 0.025 g/l and 0.060 g/l, respectively for lactic, acetic, tartaric, malic, succinic and citric acids. Validation of the method was carried out by the standard additions method, the recoveries ranged from 91.4 to 103.0%. The precision of the method was also evaluated and reported a CV% as less than 3.1%. Organic acid profiles of citrus fruit, pineapple and apple natural juices (home-made), commercial juices and nectars were established.     

Analytical Tools Used to Distinguish Chemical Profiles of Plants Widely Consumed as Infusions and Dietary Supplements: Artichoke,Milk Thistle,and Borututu

Artichoke, borututu, and milk thistle are three medicinal plants widely consumed as infusions or included in dietary supplements (e.g., pills and syrups). Despite the high consumption of these plants, studies on their nutritional value and primary metabolites are scarce, with their composition in secondary metabolites such as phenolic compounds being only reported. Therefore, their nutritional value was assessed, and analytical tools (liquid and gas chromatography coupled to different detectors) were used to distinguish their chemical profiles according to their hydrophilic (sugars and organic acids) and lipophilic (fatty acids and tocopherols) components. Chromatographic techniques are important analytical tools used in the identification and quantification of several molecules, also being a standard requirement to distinguish different profiles. Borututu gave the highest energetic value with the highest content of carbohydrates and fat, sucrose and total sugars, shikimic and citric acids, and α-, β-, δ-, and total tocopherols. Artichoke had the highest ash and protein contents, oxalic acid, SFA (mainly palmitic acid), and γ-tocopherol, and also the best n-6/n-3 ratio. Milk thistle showed the highest levels of fructose and glucose, quinic acid and total organic acids, and PUFA, mainly linoleic acid, and the best PUFA/SFA ratio. The hydrophilic compounds identified in the studied plants, mostly sugars, are responsible for the energy contribution of their widely consumed infusions. Otherwise, the bioactivity of lipophilic compounds, namely, unsaturated fatty acids and tocopherols, is lost in those preparations but can be recovered in dietary supplements based on the plants. As far as we know, this is the first report on the detailed composition of the three plants regarding their nutritional features.     

Dynamic cell fractionation and transportation using moving dielectrophoresis

This study presents a new cell manipulation method using a moving dielectrophoretic force to transport or fractionate cells along a microfluidic channel. The proposed moving dielectrophoresis (mDEP) is generated by sequentially energizing a single electrode or an array of electrodes to form an electric field that moves cells continuously along the microchannel. Cell fractionation is controlled by the applied electrical frequency, and cell transportation is controlled by the interelectrode activation time. The applicability of this method was demonstrated to simultaneously fractionate and transport Saccharomyces cerevisiae yeast cells, both viable and nonviable, by operating at conditions under which the cells were subjected to positive and negative dielectrophoresis, respectively. Compared to the conventional dielectrophoresis (cDEP and traveling wave dielectrophoresis (twDEP), moving dielectrophoresis allows cells to be separated on the basis of the real part of the Clausius-Mossotti factor, as in cDEP, but yet allows the direct transportation of separated cells without using fluid flow, as in twDEP. This dielectrophoresis technique provides a new way to manipulate cells and can be readily implemented on programmable multielectrode devices.     

TaqMan real-time PCR for the detection and quantitation of pork in meat mixtures

A rapid and highly specific real-time quantitative PCR, based on the amplification of a fragment of the mitochondrial 12S ribosomal RNA gene (rRNA), has been developed for the quantitation of pork (Sus scrofa) in binary pork/beef muscle mixtures. The method combines the use of pork-specific primers, that amplify a 411bp fragment from pork DNA, and mammalian-specific primers amplifying a 425-428bp fragment from mammalian species DNA, which are used as endogenous control. An internal fluorogenic probe (TaqMan), that hybridizes in the "pork-specific" and also in the "mammalian" DNA fragments is used to monitor the amplification of the target gene. A comparison of the cycle number (C(t)) at which mammalian and pork-specific PCR products are first detected, in combination with the use of reference standards of known pork content, allows the determination of the percentage of pork in a mixed sample. Analysis of experimental pork/beef muscle binary mixtures demonstrated the specificity and sensitivity of the assay for detection and quantitation of pork in the range 0.5-5%.     

Dietary procyanidins lower triglyceride levels signaling through the nuclear receptor small heterodimer partner

Hypertriglyceridemia is an independent risk factor in the development of cardiovascular diseases, and we have previously reported that oral administration of a grape seed procyanidin extract (GSPE) drastically decreases plasma levels of triglycerides (TG) and apolipoprotein B (ApoB) in normolipidemic rats, with a concomitant induction in the hepatic expression of the nuclear receptor small heterodimer partner (NR0B2/SHP). Our objective in this study was to elucidate whether SHP is the mediator of the reduction of TG-rich ApoB-containing lipoproteins triggered by GSPE. We show that GSPE inhibited TG and ApoB secretion in human hepatocarcinoma HepG2 cells and had and hypotriglyceridemic effect in wild-type mouse. The TG-lowering action of GSPE was abolished in HepG2 cells transfected with a SHP-specific siRNA and in a SHP-null mouse. Moreover, in mouse liver, GSPE downregulated several lipogenic genes, including steroid response element binding protein 1c (SREBP-1c), and upregulated carnitine palmitoyltransferase-1A (CPT-1A) and apolipoprotein A5 (ApoA5), in a SHP-dependent manner. In HepG2 cells GSPE also inhibited ApoB secretion, but in a SHP-independent manner. In conclusion, SHP is a key mediator of the hypotriglyceridemic response triggered by GSPE. This novel signaling pathway of procyanidins through SHP may be relevant to explain the health effects ascribed to the regular consumption of dietary flavonoids.     

Influence of grape variety and their phenolic composition on vasorelaxing activity of young red wines

Red wine has been reported to exert beneficial effects in preventing cardiovascular diseases probably due to their polyphenols constituents. The vasorelaxant capacity and phenolic composition of four monovarietal young red wines Merlot, Tempranillo, Garnacha and Cabernet-Sauvignon obtained from grapes from 2004 vintage, cultivated under the same climatic and agricultural conditions, wine-making technology and storage conditions, were determined using vascular reactivity assay in rat aortic and high-performance liquid chromatography, respectively. Primary cultures of vascular smooth muscle cells from Wistar rats were also used to evaluate their survival prevention activity as well. Every studied wine present vasorelaxation effect, but the higher value corresponds to the analyzed Merlot wine, especially rich in phenolic compounds, mainly catechins and oligomeric proanthocyanidins and anthocyanin glycosides. This study further demonstrated previous investigations about the phenolic composition of wines relation with their vasorelaxation activity.     

Changes of health-related compounds throughout cold storage of tomato juice stabilized by thermal or high intensity pulsed electric field treatments

The effect of high intensity pulsed electric fields (HIPEF) processing (35 kV/cm for 1500 μs in bipolar 4-μs pulses at 100 Hz, with an energy density of 8269 kJ/L) on the main bioactive compounds and antioxidant capacity of tomato juice was investigated and compared to heat pasteurization (90 °C for 1 min or 30 s) having the fresh juice as a reference. HIPEF and heat treated tomato juices showed higher lycopene and lower vitamin C levels than the untreated juice. However, no significant changes in the total phenolic content and antioxidant capacity were observed between treated and fresh juices just after processing. Lycopene, vitamin C and antioxidant capacity of both treated and untreated juices decreased exponentially during storage following a first order kinetics (R² = 0.763–0.987), whereas tomato juices maintained their initial phenolic content. HIPEF-treated tomato juice maintained higher lycopene and vitamin C content than the thermally treated juices during the storage time. Hence, the application of HIPEF may be appropriate to achieve nutritious and fresh like tomato juice.

Industrial relevance

HIPEF processing can lead to tomato juice with higher nutritional value than that thermally processed. HIPEF-treated (35 kV/cm for 1500 μs with 4-μs bipolar pulses at 100 Hz, energy input of 8269 kJ/L) tomato juice shows greater lycopene, vitamin C and antioxidant capacity just after the treatment and during the storage time than heat treated (90 °C­30 s and 90 °C­60 s) tomato juice. Therefore, HIPEF technology is a feasible alternative to thermal treatment to obtain tomato juice with a high presence of health-related compounds.