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Ten different barley cultivars and their corresponding malts were used to obtain different fractions. Phenolics extracted belonged to free, soluble esters and insoluble‐bound fractions. Total phenolic content (TPC) of the free fraction, as measured according to the Folin‐Ciocalteu method, ranged from 37.7 to 167.2 mg gallic acid equiv/kg of dried material (GAE/kgdw) for barley and between 34.1 and 72.3 mg GAE/kgdw for malt. The bound phenolic content ranged from 210.3 to 320.5 and between 81.1 and 234.9 mg GAE/kgdw for barley and malt, respectively. The contribution of bound phenolics to the TPC was significantly higher than that of free and esterified fractions. Catechin and ferulic acid, quantified by high performance liquid chromatography with diode array detector (HPLC‐DAD), were the most abundant phenolics in the free and bound fractions, respectively. The p‐coumaric acid content was lower in hulless genotypes, as compared to hulled genotypes, showing that it is mainly concentrated in the hull. The antioxidant activities of the phenolic fractions were investigated using the radical scavenging assay (DPPH) and ferricyanide reducing power. The bound phenolics demonstrated a significantly higher antioxidant capacity compared to the free and esterified phenolics. During the malting process, a significant decrease of the bound phenolics was observed with a corresponding increase of the esterified fraction.
J. Inst. Brew. 114(2), 150-159, 2008Hot water (45°C) extracts of ten barley varieties and their corresponding malts were analyzed in terms of antioxidant activity. The ferric reducing antioxidant power (FRAP) and radical scavenging activity (ABTS), ranged from 0.23-0.45 mg GAE/g dw for malt and 0.12-0.25 mg GAE/g dw for barley. The hull-less malt KM 1910 was the variety with the best antioxidant properties, whereas the highest antioxidant capacity for barley was detected for the variety Merlin. A significant positive correlation between the methods FRAP, ABTS and ITT was found (p < 0.01). The influence of fertilization (20 kg N/ha) on barley antioxidant capacity was studied. The results obtained suggest that the impact of fertilization was not evident and that it depends significantly on barley genotype. The total polyphenol content, as measured according to Folin-Ciocalteu's method, ranged from 0.6-2.9 mg GAE/g dw and correlated positively with all the antioxidant methods used (p < 0.01). Free phenolic compounds were measured by HPLC with a CoulArray detector. The dominant phenolic compound was ferulic acid and its content ranged from 12.5-21.9 and 7.8-56.1 μg/g dw for barley and malt, respectively. The content of catechin ranged from 11.0-17.0 μg/g dw in barley and 0.9-12.1 μg/g dw in malt.
The determination of polyphenols by spectrophotometric detection is complicated due to their low concentrations in beer. The beer samples have to be pre-concentrated before using the spectrophotometric detection for their quantification. An analytical method based on solid-phase extraction (SPE) and followed by high performance liquid chromatographic separation with diode-array detection is used for the determination of free gallic, protocatechuic, caffeic, p-coumaric, ferulic and salicylic acids, of (+)-catechin, (–)-epicatechin, and quercetin. These phenolic compounds participate in colloidal and sensory stability of beer. Six different SPE cartridges were tested and three different types of elution with the most appropriate solvents (acetonitrile, acetone and methanol) were used. The performance of the HPLC method was assessed by the evaluation of parameters such as absolute recovery, relative standard deviation (RSD – lower than 10%), the limit of quantification (LOQ), and the limit of detection (LOD). The polyphenol content in various types of Czech beer is presented.
Cumulus expansion of the cumulus-oocyte complex is necessary for meiotic maturation and acquiring developmental competence. Cumulus expansion is based on extracellular matrix synthesis by cumulus cells. Hyaluronic acid is the most abundant component of this extracellular matrix. Cumulus expansion takes place during meiotic oocyte maturation under in vivo and in vitro conditions. Quantification and measurement of cumulus expansion intensity is one possible method of determining oocyte quality and optimizing conditions for in vitro cultivation. Currently, subjective methods of expanded area and more exact cumulus expansion measurement by hyaluronic acid assessment are available. Among the methods of hyaluronic acid measurement is the use of radioactively labelled synthesis precursors. Alternatively, immunological and analytical methods, including enzyme-linked immunosorbent assay (ELISA), spectrophotometry, and high-performance liquid chromatography (HPLC) in UV light, could be utilized. The high sensitivity of these methods could provide a precise analysis of cumulus expansion without the use of radioisotopes. Therefore, the aim of this review is to summarize and compare available approaches of cumulus expansion measurement, respecting special biological features of expanded cumuli, and to suggest possible solutions for exact cumulus expansion analysis.
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