Proanthocyanidin profile and ORAC values of Manitoba berries, chokecherries, and seabuckthorn

Six Manitoba fruits were analyzed for their phytochemical content and antioxidant activity in order to increase their production and marketability. The major proanthocyanidins (flavanols) present in whole fruit, juice, and pulp of strawberry, Saskatoon berry, raspberry, wild blueberry, chokecherry, and seabuckthorn were measured. The extraction and purification were facilitated using flash column chromatography, while separation and identification were accomplished by using HPLC and LC-MS techniques. The total proanthocyanidin contents varied from 275.55 to 504.77 mg/100 g in the whole fruit samples. Raspberry contained the highest content, and seabuckthorn showed the lowest content of total flavanols. The highest concentration of proanthocyanidin in juice was found in Saskatoon berry (1363.34 mg/100 mL) and the lowest value in strawberry (622.60 mg/100 mL). HPLC and LC-MS results indicated that epicatechin was the most abundant flavanol followed by B2 in the berry samples, while no catechin or B1 was detected in these fruits. A series of oligomers and polymers were detected in all samples. The recovery percentage was obtained from the ratio of the unspiked samples to the area of spiked samples. Monomers, dimers, oligomers, and polymers gave recovery ranges of 83-99%. The lipophilic and hydrophilic antioxidant capacities of whole fruit, juice, and pulp extracts were measured by the oxygen radical absorbance capacity (ORAC) procedure. In whole fruits, the ORAC values varied from 135 to 479 mg/100 g TE in the MeOH fraction. The corresponding ORAC values varied from 115.30 to 733.15 mg/100 g for the acetone fraction. In juice, all berries showed the same antioxidant capacity (P > 0.05) (133.0-312.0 mg/100 g) in the MeOH fraction, with the exception of raspberry (603.0 mg/100 g). Overall, MeOH fractions mainly contained monomers and dimers with smaller amounts of oligomers and polymers when compared to the acetone fractions. Acetone fractions mainly contained polymers and some oligomers. Although acetone fractions contained a higher quantity of total proanthocyanidins, their antioxidant capacities were lower than MeOH fractions.     

Recovery of Salmonella from shell eggs.

A preenrichment procedure and a direct selective enrichment procedure were compared for recovery of Salmonella artificially inoculated into liquid whole egg, egg yolk, and egg albumen. For liquid whole egg and egg yolk, the 2 procedures were comparable. With egg albumen, however, preenrichment in lactose broth gave significantly higher recoveries than did direct selective enrichment in either selenite cystine or tetrathlonate broths. The lactose preenrichment procedure was used to determine the survival of S. enteritidis in egg yolk and egg albumen over a period of 7 days. As shown by most probably number determinations, counts of S. enteritidis inoculated into egg albumen decreased by 3 log units, whereas those in egg yolk did not change significantly. It is recommended, therefore, that only the egg yolk be examined for this pathogen. In a comparison of 5 different preenrichment media (lactose broth, brain heart infusion broth, trypticase soy broth, buffered peptone water, and nutrient broth), lactose broth was somewhat less productive than the other 4 media for the recovery of Salmonella from egg yolks. Trypticase soy broth gave the highest recovery.     

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.     

Assessment of egg nutrient compositional changes and residue in eggs,tissues, and excreta following oral administration of atorvastatin to laying hens

Laying hens were fed a control diet alone or with 0.06 g of atorvastatin, a synthetic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, per 100 g of diet for 20 days. Compared to controls, egg yolks from treated hens contained greater amounts of amino acids and reduced levels of total fatty acids and cholesterol. In contrast, egg albumen amino acid contents were unaffected by dietary treatments. In a residue study, seven hens each received a single oral dose of approximately 20 microCi of [(14)C]atorvastatin. Approximately 71% of the radioactivity was recovered in the excreta and liver, whereas virtually no radioactivity was detected in kidney, heart, muscle, bile, plasma, or egg albumen at 15 days postdosing. Yolk radioactivity peaked at 4 days postdosing in six of the seven birds and was absent in eggs laid after day 10. Reminiscent of that of certain antibiotic drugs, the atorvastatin egg residue pattern appeared to coincide with the physiological pattern of daily yolk accretion within the ovary.     

Industrial glycerol as a supplementary carbon source in the production of beta-carotene by Blakeslea trispora

The dynamics of industrial types of glycerol as a supplementary carbon source to glucose for beta-carotene production by Blakeslea trispora was investigated in batch cultures. The growth kinetics, cellular lipid accumulation-degradation, substrate assimilation, and beta-carotene production were clearly dependent on the level of addition of pure glycerol. The highest beta-carotene production (15.0 mg/g of dry biomass) was obtained at an initial glycerol concentration of 60.0 g/L. Substitution of pure glycerol by the nonpurified soap byproduct did not inhibit cell growth. Conversely, partial purification of the biodiesel byproduct by removing methanol and fatty acids was unavoidable for cell growth. Both types of industrial glycerol stimulated beta-carotene synthesis more than 10 (soap byproduct) and 8 times (biodiesel byproduct) compared to control medium. The maximum beta-carotene contents were 10 and 8 mg/g of dry biomass, respectively, and its relative content in the carotenoid fraction was 86-88%.     

Amino-N compounds found in soil organic matter hydrolysates of a loamy sand using an immobilized protease reactor column

Summary Soil organic matter (OM) from seven different fertility plots of a loamy sand was extracted and fractionated into high- and low-molecular-weight (HMW, LMW) fractions using gel filtration. The fractions were acid-hydrolyzed to determine the amino sugar and amino acid contents. The same fractions were hydrolyzed with an immobilized protease reactor column. Reverse-phase high-performance liquid chromatography (HPLC) was used to identify the soil amino-N compounds. With the HMW fraction as substrate, the enzyme released less than 1% of 11 amino-N compounds determined by acid hydrolysis. Phenylalanine and leucine, however, were recovered in quantities of 2% and 4%, respectively. Immobilized protease hydrolysis of the LMW fraction recovered considerably more amino-N compounds compared with acid hydrolysis of the same fractions. Each system of hydrolysis produced some amino-N compounds not found in the other. We conclude that an immobilized enzyme reactor column will allow a researcher to perform time-course hydrolysis, so that hydrolysis intermediates, e.g. peptides, can be separated and identified.     

Separation of ovomucin subunits by gel filtration: enhanced resolution of subunits by using a dual-column system.

An improved procedure involving a dual-column gel-filtration system for the separation of ovomucin subunits is described. Ovomucin, prepared from thick egg albumin by precipitation, was separated in the presence of sodium dodecyl sulfate (SDS) and beta-mercaptoethanol by using two Sephacryl S-400 HR gel-filtration columns connected in series. The separation resulted in the isolation of eight peaks. The amino acid compositions and SDS-polyacrylamide gradient gel electrophoretic pattern of each peak were determined.     

Determination of free flavan-3-ol content in barley (Hordeum vulgare L.) air-classified flours: comparative study of HPLC-DAD/MS and spectrophotometric determinations

The determination of free flavan-3-ol compounds in barley flours (cv. Gotic) and two resulting milling fractions (fine fraction 57% and coarse fraction 43%, w/w) obtained by air classification of dehulled grain is herein described. The determinations were carried out using reversed phase high-performance liquid chromatography coupled with both diode array detection and ESI-MS compared with conventional spectrophotometric determinations (total phenolic compounds by the Folin-Ciocalteu method and free radical scavenging activity with the DPPH assay). Significant correlations among the HPLC quantification, the spectrophotometric data, and the antioxidant capacity of extracts were revealed by Pearson's analysis. Nine flavan-3-ols were identified by HPLC-MS. Catechins and their derivatives were found to make a substantial contribution to the antioxidant power of extracts. The coarse fraction showed larger concentrations of flavan-3-ols (221%) with respect to the fine fraction. This was confirmed by the antioxidant activity of the analyzed flours. The coarse fraction showed the greatest antioxidant activity (1200.1 +/- 66.2 micromol of Trolox equiv/100 g of flour) with respect to whole meal and fine fraction (1025.9 +/- 18.3 and 761.7 +/- 55.3 micromol of Trolox equiv/100 g of flour, respectively).     

Polyphenolic profiles and antioxidant activities of heartnut (Juglans ailanthifolia Var. cordiformis) and Persian walnut (Juglans regia L.)

The polyphenolic compositions of three heartnut (Juglans ailanthifolia var. cordiformis) varieties (Imshu, Campbell CW1, and Campbell CW3) were examined and compared with those of two Persian walnut (Juglans regia L.) varieties (Combe and Lake). The nuts were defatted, extracted, and separated into three different fractions, the free phenolic acid (FPA), acid-hydrolyzable phenolic acid (AHPA), and bound phenolic acid (BPA) fractions. The total phenolic contents (TPCs) in both FPA and AHPA of the Persian walnuts were significantly higher (P < 0.001) than those of the heartnuts, but not in the BPA (P = 0.20). LC-ESI-MS(n)() studies revealed that except for the FPA fraction, the major polyphenolics in both heartnut and Persian walnut were ellagic acid and valoneic acid dilactone. Persian walnuts contained an average of 0.29 and 1.31 mg of ellagic acid/g nut in the 80% methanol extractable fractions FPA and AHPA, respectively. Heartnuts contained an average of 0.16 and 0.60 mg of ellagic acid/g nut in the respective fractions. Bound ellagic acid in the residue was 0.93 and 0.70 mg/g of nut in the Persian walnut and in the heartnut, respectively. Valoneic acid dilactone was tentatively identified and quantified as milligrams of ellagic acid equivalent per gram of nut. These components were found to contribute to the strong total antioxidant activities measured using ferric reducing antioxidant power and photochemiluminescence methods.     

Effectiveness of the Iodide Ligand Along with two Surfactants on Desorbing Heavy Metals from Soils

Used with one of two surfactants (SDS, an anionic surfactant, and Triton X-100, a nonionic surfactant), the ligand, I^? was evaluated as a washing agent for the desorption of Cd from naturally and artificially contaminated soils. Increasing amounts of the ligand, I^?, with a surfactant, specifically removes higher levels of Cd but not Cu, Zn and Pb. After seven washings, the ligand, I^? with the nonionic surfactant, Triton X-100, removed 65 and 90% of the Cd from soils I and II, containing respectively, to 15 and 1275 mg of Cd/kg. The ligand, I^?, and the anionic surfactant, SDS, removed 35 and 70% of the Cd from soils I and II, respectively. Before washing, the carbonate fraction of soil I contained the most Cd (48%) while the exchangeable and carbonate fractions of soil II contained 29 and 33% of the total Cd, respectively. For soil I, SDS with/ without the ligand desorbed Cd mainly from the carbonate and oxide fractions, while only Triton X-100 with ligand I^? could remove Cd from the exchangeable fraction. For soil II, Cd was desorbed from the exchangeable fraction only when either surfactant was used in combination with the ligand. Thus, a surfactant with ligand can extract specific heavy metals from soils and selective sequential extraction is useful in identifying which fraction can be targeted by the surfactant – ligand agent.     

Distribution of total 14C residue in egg yolk, albumen, and tissues following oral [14C]sulfamethazine administration to hens

The distribution of total 14C residues was studied in egg yolk and albumen after administration of either single or multiple oral dosages of [14C]sulfamethazine (SMZ). One day after a single dose of [14C]SMZ (121 mg of sulfamethazine, 2.42 x 10(7) dpm), the 14C residue concentration peaked in egg albumen and egg yolk with the concentration in the former >4-fold greater than in the latter. Three days postdose, the 14C residue concentration in the yolk was approximately 7-fold higher than in the egg albumen. A multiple dose of [14C]SMZ containing sulfamethazine mass equivalent of an average therapeutic dose (282 mg, 2.9 x 10(7) dpm) for chickens was also administered orally for six consecutive days to hens. A significantly reduced level of egg production was observed during the medication, and most of the hens stopped laying eggs after the last dose. The 14C residue concentrations peaked on the last day (sixth) of medication in egg albumen and yolk. The 14C residue concentrations were also measured in liver, muscle, blood, and plasma of chickens sacrificed at 1, 24, 48, and 72 h after the last dose. Highest concentrations of 14C residue were accumulated in liver followed by, in decreasing order, blood, plasma, and muscle.     

Effects of urease and nitrification inhibitors on the efficient use of urea for pastoral systems

Abstract

The humic substances contained in an animal organic waste were extracted and the total extract separated into three humic fractions with different molecular weights (low, F₁ <10³; medium, F₂, with molecular weights ranging from 10³ to 10⁴; and high, F₃ >10⁴). The C content was highest in F₂, the same fraction also showing the lowest N content. The molecular weight of the humic fractions influenced the electrical conductivity, the highest molecular weight resulting in the lowest degree of electrical conductivity. Membrane-controlled ultrafiltra-tion (the method used to separate the various fractions from the whole extract) was also suitable for purifying such enzymes as phosphatase and β-glucosidase: the total activity obtained from the three fractions was considerably greater than that determined in the whole extract, Pyrolysis-gas chromatography (Py-GC) applied to the whole extract and humic fractions showed that in the F₃ fraction (highest molecular weight) benzene was the major fragment while furfural was the major fragment of F₁ (lowest molecular weight). For this reason, the humification index benzene/toluene indicates that the fraction with the highest molecular weight was the most humified while the furfural/pyrrole ratio indicates that the fraction with the lowest molecular weight was the most degradable. The whole extract and the fraction F₁ had a negative effect on seed germination when the concentration was equivalent to 100 mg kg^?1 of C, while the germination index was higher than that of the control when only 10 mg kg^?1 were used. The F₂ fraction had a positive effect on germination regardless of the concentration used. When 10 mg kg^?1 of C of the humic substances studied were added to the nutrient solution for growth experiments with maize plants, F₃ led to increases in root weight and F₂ led to increases in shoot weight. An inhibitor effect was observed for fraction F₁.     

Studies of selected plant raw materials as alternative sources of triterpenes of oleanolic and ursolic acid types

The qualitative and quantitative evaluation of triterpene aglycones of saponin fractions isolated from vegetative and generative organs of three Silphium species, Silphium perfoliatum, Silphium trifoliatum, and Silphium integrifolium, as compared to materials used in the herbal industry such as Panax quinquefolium root and Calendula officinalis flower, was performed. The analyses revealed that triterpene aglycones of saponins isolated from tested Silphium and Calendula species were oleanolic acid and ursolic acid. It was found that Panax roots contained only the aglycone of oleanolic acid within the triterpene saponin group. The leaves of Silphium harvested in May were characterized by the highest content of oleanolic acid-They contained 17.03 mg/g dry weight of the triterpenic acid, on average. The seasons before flowering and at the beginning of that stage appeared to be the most efficient periods for leaf collection in reference to triterpene aglycone contents in plant yield. Moreover, it was found that inflorescences of S. trifoliatum and S. integrifolium contained oleanolic acid in amounts of 22.05 and 17.95 mg/g dry weight respectively, whereas Calendula flowers contained 20.53 mg/g dry weight. The oleanolic acid content in Panax roots was 3.15 mg/g dry weight. Ursolic acid most abundantly occurred in S. integrifolium and S. trifoliatum at concentrations of about 14.98 mg/g dry weight in leaves harvested before flowering (June) and to 15.50 mg/g dry weight in leaves collected during flowering.     

Importance of insoluble-bound phenolics to antioxidant properties of wheat

Two commercial samples of soft (70% Canadian Eastern soft red spring and 30% Canadian Eastern soft white winter) and hard (90% Canadian western hard red spring and 10% Canadian Eastern hard red winter) wheats were used to obtain different milling fractions. Phenolics extracted belonged to free, soluble esters and insoluble-bound fractions. Soluble esters of phenolics and insoluble-bound phenolics were extracted into diethyl ether after alkaline hydrolysis of samples. The content of phenolics was determined using Folin-Ciocalteu's reagent and expressed as ferulic acid equivalents (FAE). The antioxidant activity of phenolic fractions was evaluated using Trolox equivalent antioxidant capacity, 2,2-diphenyl-1-picrylhydrazyl radical scavenging, reducing power, oxygen radical absorbance capacity, inhibition of oxidation of human low-density lipoprotein cholesterol and DNA, Rancimat, inhibition of photochemilumenescence, and iron(II) chelation activity. The bound phenolic content in the bran fraction was 11.3 +/- 0.13 and 12.2 +/- 0.15 mg FAE/g defatted material for hard and soft wheats, respectively. The corresponding values for flour were 0.33 +/- 0.01 and 0.46 +/- 0.02 mg FAE/g defatted sample. The bound phenolic content of hard and soft whole wheats was 2.1 (+/-0.004 or +/-0.005) mg FAE/g defatted material. The free phenolic content ranged from 0.14 +/- 0.004 to 0.98 +/- 0.05 mg FAE/g defatted milling fractions of hard and soft wheats examined. The contribution of bound phenolics to the total phenolic content was significantly higher than that of free and esterified fractions. In wheat, phenolic compounds were concentrated mainly in the bran tissues. In the numerous in vitro antioxidant assays carried out, the bound phenolic fraction demonstrated a significantly higher antioxidant capacity than free and esterified phenolics. Thus, inclusion of bound phenolics in studies related to quantification and antioxidant activity evaluation of grains and cereals is essential.     

In vitro bioavailability of phenolic compounds from five cultivars of frozen sweet cherries (Prunus avium L.)

The bioavailability of phenolic compounds from five cultivars of frozen sweet cherries was assessed by a digestion process involving pepsin-HCl digestion (to simulate gastric digestion) and pancreatin digestion with bile salts (to simulate small intestine conditions) and dialyzed to assess serum- and colon-available fractions. After pepsin digestion, the % recovery of total phenolics, relative to the original starting material, increased, whereas the % anthocyanins did not change. Following pancreatic digestion and dialysis, the total phenolics in the IN (serum-available) fraction was about 26-30% and the OUT (colon-available) fraction was about 77-101%. The anthocyanin content in the IN fraction was 15-21%, and in the OUT fraction, it was 52-67%. Skeena, Lapins, and Sweetheart cultivars contained higher levels of total phenolics and anthocyanins, which resulted in higher concentrations of these compounds in the IN and OUT fractions. The potential bioavailability of phenolic compounds was also assessed in Bing and Lapins cultivars at three ripening stages. Immature cherries had higher % total phenolics in the IN fraction than mature or overmature cherries. However, immature cherries had the lowest concentrations of these compounds, making the actual bioavailable amounts of these compounds lower than for mature and overmature fruit. High-performance liquid chromatography analysis of Lapins cherries at three maturity stages confirmed the results obtained using spectrophotometric methods for total phenolics and anthocyanins.     

Evidence of peptides in low-molecular-weight fractions of soil organic matter

Summary Organic matter was extracted from three soils, a Berwick sandy loam, a Franklin loamy sand, and a Cumberland silty loam. The extracts were separated into high (>8000 daltons) and low-molecular-weight (<8000 daltons) fractions using gel filtration. Reverse-phase high performance liquid chromatography at 214 nm was used to separate the peptides into low-molecular-weight fractions. Peptide samples were collected with an integrated fraction collector and hydolyzed with an immobilized protease column reactor. High performance liquid chromatography with fluorescence detection was used to determine the amino-acid contents of the collected samples. The results indicated that peptide intermediates are present in soil size fractions. Greater quantities of several amino acids were released from the peptide hydrolyzates of the Berwick sandy loam and Franklin loamy sand, compared with the Cumberland silty loam, an uncultivated soil. These findings indicate that organic intermediates (e.g., peptides) are more prevalent in biologically active soils than in relatively inert soils.     

Xanthophylls in commercial egg yolks: quantification and identification by HPLC and LC-(APCI)MS using a C30 phase

The xanthophylls lutein and zeaxanthin have attracted a lot of interest since it was presumed that an increased nutritional uptake may prevent adult macula degeneration (AMD). Although egg yolks serve as an important dietary source of lutein and zeaxanthin, data on xanthophyll concentrations in commercial egg yolks are not available. Thus, an high-performance liquid chromatography-diode array detector (HPLC-DAD) method was developed allowing for simultaneous separation of eight xanthophylls used to fortify poultry feed. Peak identification was carried out by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [LC-(APCI)MS]. Egg yolks of four types of husbandry (seven batches each) were examined. Lutein and zeaxanthin were the predominant xanthophylls in egg yolks produced in accordance with ecological husbandry (class 0) because the concentrations of these xanthophylls ranged from 1274 to 2478 microg/100 g and from 775 to 1288 microg/100 g, respectively. Analysis of variance (ANOVA) proved that both mean lutein and mean zeaxanthin concentrations of eggs from class 0 were statistically discriminable from mean lutein and zeaxanthin concentrations from eggs of all other classes (P < 0.01). Mean concentrations of synthetic xanthophylls in eggs of classes 1 (free range), 2 (barn), and 3 (cage) were as follows: canthaxanthin, 707 +/- 284 microg/100 g; beta-apo-8'-carotenoic acid ethyl ester, 639 +/- 391 microg/100 g; and citranaxanthin, 560 +/- 231 microg/100 g. Experiments with boiled eggs proved that beta-apo-8'-carotenoic acid ethyl ester was the xanthophyll with the highest stability, whereas lutein was degraded to the largest extent (loss of 19%). Detailed knowledge about the xanthophyll amounts in eggs is indispensable to calculate the human uptake.     

Broth from canned clams is suitable for use as an excellent source of free vitamin B12

Vitamin B(12) was assayed and characterized in the broth of canned clams (boiled plain). The broth contained considerable amounts of vitamin B(12) (2.7-14.1 μg/100 g, 1.3-6.7 μg/can). HPLC and LC/ESI-MS/MS chromatograms demonstrated that the clam broth contained true vitamin B(12). Gel filtration experiments indicated that most (72%) of the vitamin B(12) found in the broth was recovered in free vitamin B(12) fractions. These results indicate that the clam broth would be suitable for use an excellent source of free vitamin B(12) for elderly persons with food-bound vitamin B(12) malabsorption.     

Potential antioxidant capacity of sulfated polysaccharides from the edible marine brown seaweed Fucus vesiculosus

Fucus vesiculosus was sequentially extracted with water at 22 degrees C (fraction 1 (F1)) and 60 degrees C (F2), and with 0.1 M HCl (F3) and 2 M KOH (F4) at 37 degrees C. Soluble fractions (42.3% yield) were composed of neutral sugars (18.9-48 g/100 g), uronic acids (8.8-52.8 g/100 g), sulfate (2.4-11.5 g/100 g), small amounts of protein (< 1-6.1 g/100 g), and nondialyzable polyphenols (0.1-2.7 g/100 g). The main neutral sugars were fucose, glucose, galactose, and xylose. Infrared (IR) spectra of the fractions showed absorption bands at 820-850 and 1225-1250 cm(-1) for sulfate. F1, F2, and F4 also exhibited an absorption band at 1425 cm(-1), due to uronic acids, and their IR spectra resembled that of alginate. F3 had an IR spectrum similar to that of fucoidan with an average molecular weight of 1.6 x 10(6) Da, calculated by molecular exclusion high-performance liquid chromatography. The presence of fucose in this polysaccharide was confirmed by (1)H NMR spectroscopy. This fraction showed the highest potential to be antioxidant by the ferric reducing antioxidant power (FRAP) assay, followed by the alkali- and water-soluble fractions. Sulfated polysaccharides from edible seaweeds potentially could be used as natural antioxidants by the food industry.