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Phenolic compounds were extracted from buckwheat and buckwheat groats into 80% acetone (v/v). The crude extracts were separated with Sephadex LH-20 column chromatography into low molecular weight phenolic compounds and tannin fractions. The tannin fractions were characterised by colour reaction with vanillin/HCl reagent, ability for protein precipitation as well as SE-HPLC method. Chelation of Cu(II) and Zn(II) by tannin fractions was determined by reaction with tetramethylmurexide (TMM) whereas Fe(II) chelation was investigated by forming complexes with ferrozine. The tannin fractions obtained from buckwheat and buckwheat groats gave similar results in vanillin test; A500/mg was 1.04. Also results obtained using precipitation method were comparable. SE-HPLC chromatograms showed that in tannin fraction of buckwheat groats polymers with molecular weight ≥ tannic acid were present. On the chromatogram of buckwheat tannins peaks were shifted to lower molecular weight compounds. Tannins of buckwheat were stronger chelators of all metal ions tested (Fe(II), Cu(II) and Zn(II)) than tannins of buckwheat groats. Ions of copper were chelated the most effectively; 1 mg of buckwheat and buckwheat groats tannin fractions added per assay bound 86.0% and 72.9% of Cu(II), respectively. Buckwheat tannins used in the test at a level of 2.5 mg/assay bound approximately twice more Fe(II) than the same addition of buckwheat groats tannins. Zn(II) was chelated to the lowest extent, 17.2% was bound by buckwheat tannins and 13.5% by buckwheat groats tannins (5 mg of tannin/assay).
Z odtłuszczonych nasion soi wyekstrahowano związki fenolowe 80% (v/v) roztworem metanolu. Uzyskany ekstrakt rozdzielono na sześć frakcji (I- VI) stosując chromatografią kolumnową na żelu Sephadex LH-20 z metanolem jako fazą ruchomą. W uzyskanych frakcjach oznaczono zawartość fenoli ogółem i zarejestrowano widma UV. Frakcje scharakteryzowano również za pomocą chromatografii cienkowarstwowej na żelu krzemionkowym. Aktywność przeciwutleniającą mierzono w układzie modelowym z ß-karo- tenem i kwasem linolenowym.
Z ziarniaków pszenicy otrzymano ekstrakt związków fenolowych stosując ekstrakcję 80% metanolem i niskociśnieniową chromatografię kolumnową na żelu RP-18. Związki fenolowe zawarte w tak otrzymanym ekstrakcie analizowano następnie jakościowo metodą RP-HPLC-DAD w systemie gradientowym. Otrzymany chromatogram charakteryzował się obecnością 8 głównych pików. Jeden z nich pochodził od kwasu sinapowego. Widma UV-DAD pozwalają stwierdzić, że pozostałe związki fenolowe były pochodnymi kwasu sinapowego.
Roasted sesame seeds, pumpkin seeds, sunflower seeds, soy beans, and wheat germs were obtained from Ekoprodukt company (Częstochowa, Poland). Phenolic compounds were extracted with 80% aqueous methanol. The extract of roasted sunflower seeds was characterised by a higher content of total phenolics (158 mg/g). The Total Antioxidant Capacity was the highest in the extract of roasted sunflower seeds (0.478 mmol Trolox/g) followed by the extract of roasted wheat germs (0.066 mmol Trolox/g). The extracts of roasted sunflower seeds and wheat germs were strong scavengers of DPPH radical. Their reducing power was strong as well.
Lentil meal proteins were treated by chymotrypsin. Hydrolysis was controlled with the pH-stat method. Degree of hydrolysis (DH) was evaluated and after 120 min of the process amounted to 13%. SDS-PAGE and SE-HPLC methods were used to study molecular weight distribution of lentil meal proteins and their chymotryptic hydrolysates of DH 2%, 4%, 8% and 12%. Bands in the range of 21–66 kDa were predominant on the electrophoregram of lentil proteins. Chymotrypsin treatment resulted in releasing the hydrolysis products of both high molecular weight (62,000; 30,500 Da) molecules and small peptides (<6,500 Da). At the first stage of hydrolysis (DH 2.0%) intermediate products are formed, which are then further hydrolysed. Chromatographic separation confirmed the results of SDS-PAGE. Larger polypeptides and unhydrolysed proteins are present in hydrolysate of DH 12% but products of hydrolysis with molecular weight of 0.5–6.5 kDa are predominant. No simple correlation between degree of hydrolysis and intensity of bitterness and astringency sensation was noticed. Bitterness of hydrolysates was not high (2.25-2.35 a.u.).
Ethanolic extracts of phenolic compounds were prepared from green teas (Camellia sinensis L.), bearberry leaves (Arctostaphylos uva-ursi L.), hazelnuts (Corylus avellana), evening primrose (Oenothera biennis) and grape seeds (Vitis vinifera L.). All crude extracts were examined for their gallic acid content by HPLC. The bearberry-leaf preparation was also fractionated by a Sephadex LH-20 column chromatographic method with 95% (v/v) ethanol and acetone:water (1:1; v/v) as mobile phases: one fraction consisted of low molecular-weight phenolics and the other of tannins. To all samples tannase was applied in an effort to liberate gallic acid from the phenolic esters and hydrolysable tannins. The content of free gallic acid in the extracts ranged from 1 to 15 mg/g with the highest quantity being found in evening primrose. The content of gallic acid liberated by tannase ranged from 5 to 309 mg/g with the highest content being found in the tannin fraction from the bearberry-leaf extract.
Chromatografia kolumnowa na żelu krzemionkowym z eterem etylowym jako fazą ruchomą pozwala oczyszczać ekstrakt surowego likopenu w stopniu zbliżonym do metody ekstrakcji do fazy stałej (SPE). Ekstrakt oczyszczony tą metodą może być wykorzystywany do badan modelowych lub do dalszego oczyszczenia na semipreparatywnej kolumnie HPLC.
Five fractions of phenolic compounds were obtained from the extract of common buckwheat seed (Fagopyrum esculentum Moench) using Sephadex LH-20 column chromatography with methanol as a mobile phase. The total phenolics content ranged from 19.8±1.5 (fraction I) to 164±2.2 mg (+)-catechin eq/g (fraction IV). The profiles of phenolic acids and flavonoids in the fractions were analysed using RP-HPLC-DAD. The antioxidant activity was tested as ABTS^+ and DPPH scavenging activity and capability to reduce the Fe(III)/2,4,6-Tris(2-pyridyl)-s-triazine complex to the ferrous form. Results were expressed as Trolox equivalent antioxidant capacity (TEAC), IC50 and ferric reducing antioxidant power (FRAP) values, respectively. The highest antioxidant activity was noted for fraction IV that was predominated by flavones. TEAC, IC50 and FRAP values were: 1.47±0.01 mmol Trolox eq/g, 0.058±0.003 mg/assay and 2.18±0.05 mmol Fe(II)/g, respectively. Rutin constituted 77.7% of the compounds identified in fraction III. The antiradical activity and reducing capability of this fraction were lower compared to fraction IV, but significantly higher than in fractions I and II. The main phenolic compounds of fractions I and II were phenolic acids (caffeic, 5-O-caffeoylquinic and p-coumaric). The antioxidant activity of fraction V was similar to that of fraction III.
Z nasion fasoli adzuki, gryki, kaszy gryczanej orzechów włoskich i laskowych oraz migdałów otrzymano ekstrakty związków fenolowych stosując 80% aceton. Zawartość fenoli ogółem i tanin w ekstraktach oznaczono metodami kolorymetrycznymi. Ekstrakty wykazywały zróżnicowaną aktywność przeciwutleniającą i antyrodnikową oraz różną zdolność do chelatowania Fe(II).
The phenolic compound profile and antioxidant potential of the false flax (Camelina sativa L.) plant, harvested at five morphological stages, that is, from the vegetative to the ripe seed-pod stage, have been investigated. False flax extracts were prepared using 80% (v/v) methanol, and the total phenolic content (TPC), the contents of the individual phenolics and antioxidant activity, measured as the Trolox equivalent antioxidant capacity (TEAC), ferric-reducing antioxidant power (FRAP), DPPH• scavenging activity and the ability to inhibit the oxidation of β-carotene-linoleic acid emulsion, were determined. The TPC of the plant, at different growth stages, ranged from 49.2 to 59.1 mg GAE/g of extract and from 1.46 to 3.10 mg GAE/g of fresh matter (FM). Four main phenolic compounds were identified (chlorogenic acid, rutin, quercetin 3-O-glucoside, and quercetin glycoside). The chlorogenic acid content and the sum of flavonoids increased in the extracts from the vegetative to the bud stage, reaching 35.9 and 49.5 mg/g of extract, respectively, and gradually decreased in the subsequent growth stages. The plant extracts at the bud and flowering stages generally had the highest antioxidant activity in the polar systems (TEAC, FRAP and DPPH assays). The ripe seed-pod stage showed the highest antioxidant potential in these conditions when the results were expressed on FM basis. The best antioxidant activity in the lipid emulsion system was shown for the false flax extracts at the flowering and ripe seed-pod stages. Our research has indicated the possibility of using the aerial part of C. sativa as a source of ingredients with protective antioxidant activity.
Z ziarniaków pszenicy uzyskano ekstrakt związków fenolowych stosując 80% metanol. Po hydrolizie zasadowej zawartość fenolokwasów oznaczano za pomocą metody HPLC w układzie odwróconych faz na kolumnie z wypełnieniem na bazie polimeru. Na chromatogramie uzyskano piki odpowiadające kwasowi p-kumarowemu, ferulowemu i sinapowemu. Czas analizy był kilkakrotnie krótszy od tego, jaki uzyskuje się na typowej kolumnie C18. W pracy zanotowano doskonałą liniowość pomiędzy stężeniem wzorca a polem powierzchni odpowiadającego mu piku. Stwierdzono, że analiza HPLC może być prowadzona przy detekcji w UV przy dł. fali 330 nm.
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