Oxidative status, in vitro iron-induced lipid oxidation and superoxide dismutase,catalase and glutathione peroxidase activities in rhea meat

Rhea (Rhea americana) muscles Obturatorius medialis (OM) Iliotibialis lateralis (IL) and Iliofibularis (I), obtained from farmed animals, were evaluated regarding their oxidative/antioxidant status. The mean level of thiobarbituric acid reactive substances (TBARS) expressed as malonaldehyde (MDA) content was of 0.84 mg MDA/kg wet tissue for the three muscles. TBARS level was significantly higher in IL than OM and I, with the two latter showing similar levels. The mean level of carbonyl proteins expressed as dinitrophenylhydrazine (DNPH) was 1.59 nmol DNPH mg⁻¹. Carbonyl protein levels were significantly different (P < 0.05) between the three muscles (IL > OM > I). Iron-induced TBARS generation was not significantly different between the three muscles at any time, nor for each muscle during the 5 h of the experiment. Superoxide dismutase activity in IL muscle was significantly higher (P < 0.05) than in I muscle. However, the difference between IL and OM muscles was not significant. The differences between the three muscles became not significant when the results were expressed by mg of protein contained in the extract, instead by g of wet tissue. No differences were found for catalase (μmol of discomposed H₂O₂ min⁻¹ g⁻¹ wet tissue or by mg of protein contained in the extract) and glutathione peroxidase (μmol of oxidized NADPH min⁻¹ g⁻¹ of wet tissue or by mg of protein contained in the extract) activities between the three muscles.     

The roles of catechins and caffeine in cream formation in a semi-fermented tea

The mechanism of cream formation in a semi-fermented tea was investigated by adding caffeine or catechins to the de-creamed tea infusions. The addition of catechins to the clarified infusion could not re-induce cream formation. However, creaming occurred again after replenishing the de-creamed tea infusion with caffeine. It was thought that the ester-form catechins which possessed a galloyl group and a hydroxylphenyl B ring acted like a claw. They could thus entrap and interact with caffeine more effectively through hydrogen bonding and hydrophobic interaction to form a large complex, resulting in creaming. © 1999 Society of Chemical Industry     

Analysis of some selected catechins and caffeine in green tea by high performance liquid chromatography

Green tea seems to have a positive impact on health due to the catechins-found as flavanols. Thus, the present study was aimed to develop a low cost reversed phase high performance liquid chromatographic (HPLC) method for simultaneous determination of flavanol contents, namely catechin (C), epicatechin (EC), epigallocatechin (EGC), epicatechin 3-gallate (ECG) and epigallocatechin 3-gallate (EGCG) and caffeine in 29 commercial green tea samples available in a Saudi Arabian local market. A C-18 reversed-phase column, acetonitrile–trifluoroacetic acid as a mobile phase, coupled with UV detector at 205 nm, was successfully used for precise analysis of the tested analytes in boiled water of digested tea leaves. The average values of N (No. of theoretical plates), HETP (height equivalent of theoretical plates) and R_s (separation factor) (at 10 μg ml⁻¹ of the catechins EC, EGC, EGCG and ECG) were 2.6 × 10³ ± 1.2 × 10³, 1.7 × 10⁻³ ± 4.7 × 10⁻⁴ cm and 1.7 ± 5.53 × 10⁻², respectively. The developed HPLC method demonstrated excellent performance, with low limits of detection (LOD) and quantification (LOQ) of the tested catechins of 0.004–0.05 μg ml⁻¹ and 0.01–0.17 μg ml⁻¹, respectively, and recovery percentages of 96–101%. The influence of infusion time (5–30 min) and temperature on the content of the flavanols was investigated by HPLC. After a 5 min infusion of the tea leaves, the average concentrations of caffeine, catechin, EC, EGC, ECG and EGCG were found to be in the ranges 0.086–2.23, 0.113–2.94, 0.58–10.22, 0.19–24.9, 0.22–13.9 and 1.01–43.3 mg g⁻¹, respectively. The contents of caffeine and catechins followed the sequence: EGCG > EGC > ECG > EC > C > caffeine. The method was applied satisfactorily for the analysis of (+)-catechin, even at trace and ultra trace concentrations of catechins. The method was rapid, accurate, reproducible and ideal for routine analysis.     

Scavenging effect of tea catechins and their epimers on superoxide anion radicals generated by a hypoxanthine and xanthine oxidase system

Catechins are a major group of polyphenolic compounds contained in abundance in green tea. Using electron spin resonance spectroscopy along with a spin‐trapping agent, the scavenging effect of tea catechins and their corresponding epimers against superoxide anion radicals generated by a hypoxanthine and xanthine oxidase reaction system was evaluated. The presence of 3′,4′,5′‐trihydroxyl groups attached to the B‐ring of the flavan skeleton of tea catechins elevated their radical‐scavenging efficiency in comparison to those with 3′,4′‐dihydroxyl groups. There were no significant differences between the four dominant tea catechins and their corresponding epimers with regard to radical‐scavenging ability. Under the different spin‐trapping agent concentrations, the sigmoid curves of reducing spin‐trapping adducts produced by tea catechins were shifted leftward, suggesting that a likely possible action of tea catechins is to scavenge superoxide anion radicals directly, not to inhibit the function of xanthine oxidase. Although caffeine is also known as a major ingredient of tea, its superoxide anion radical‐scavenging effect was much weaker than that of the catechin family. It is concluded that tea catechins and their epimers serve as powerful antioxidants for directly eliminating superoxide anion radicals, and may be useful in the prevention of diseases relating to in vivo oxidative stress. © 2000 Society of Chemical Industry     

Synthesis of superoxide dismutase,catalase and other enzymes and oxygen and superoxide toxicity during changes in oxygen concentration in cultures of brewing yeast

The physiological effects on brewing yeast, growing in semi-defined wort medium, of a sudden transition from aerobiosis to anaerobiosis were studied. Two yeast strains were examined, used for ale and lager fermentations respectively. The reverse transition (from anaerobiosis to aerobiosis) was also examined. Transitions were applied by changing the sparging gas during growth or in stationary phase, and the effects on the specific activities of certain key enzymes and on the viability of the cultures were examined. Neither type of transition led to significant changes in growth rate, the rate of ethanol production or the specific activities of alcohol dehydrogenase and pyruvate decarboxylase. The most significant change was in the specific activity of CuZn-superoxide dismutase, which showed a rapid increase in activity on transition from anaerobiosis to aerobiosis, and a decrease in activity on the reverse transition. Catalase activity in the ale yeast generally followed that of CuZn-superoxide dismutase, whereas in the lager yeast it remained unchanged by the transitions. The transition from anaerobiosis to aerobiosis caused increases in citrate synthase and Mn-superoxide dismutase, though only after a significant lag period. Aerobic to anaerobic transitions caused a decrease in Mn-superoxide dismutase activity, while citrate synthase remained unchanged. Anaerobically grown cells showed a rapid loss in viability on exposure to oxygen (5–7% in the first hour), while aerobically grown cells were unaffected. When anaerobically grown cells were exposed to 0·25 mM -potassium superoxide, there was an 8% loss of viability within 10 min, whereas aerobic cells were not affected. It is concluded that the toxic effect of oxygen is due to superoxide (or a species derived from it) and that the CuZn-superoxide dismutase (but not the Mn-isoenzyme) plays a role in protecting the cells. The de novo synthesis of the CuZn-enzyme is not always rapid enough to confer full protection.     

普洱熟茶不同溶剂萃取层中儿茶素及黄酮醇类化合物差异的研究

目的 探索普洱熟茶在不同溶剂(乙酸乙酯和正丁醇)萃取中儿茶素和黄酮醇类化合物的组成及含量的差异,构建一种有效提取普洱熟茶中儿茶素及黄酮醇类物质的方法。方法 以普洱熟茶(2008年生产)的水提物为研究对象,分别用乙酸乙酯和正丁醇溶剂萃取的萃取层为分析供试样,采用高效液相色谱法(high performance liquid chromatography, HPLC)对供试茶样中5种主要儿茶素和4种黄酮醇类化合物的含量进行了测定。结果 儿茶素、表儿茶素、表儿茶素没食子酸酯、表没食子儿茶素没食子酸酯、杨梅素、槲皮素和山奈酚主要富集于乙酸乙酯萃取层中;而芦丁与表没食子儿茶素分别富集于正丁醇层与萃取剩余层。结论 建立了一种简易、有效地分析检测普洱熟茶中儿茶素及黄酮醇类物质的提取方法,为进一步解析普洱熟茶的化学成分提供一定的方法借鉴。     

Factors affecting the levels of catechins and caffeine in tea beverage: estimated daily intakes and antioxidant activity

Increased public awareness of health benefits of green tea is generally based on the high polyphenol content of tea leaves and the resulting beverage. A number of factors, such as species, season, agronomic condition and age of the leaves, are known to affect the composition of commercial teas. In the present study the effects of factors associated with domestic preparation and analytical methods, such as brewing time, concentration, solvent and type of tea product, on levels of catechins and caffeine, antioxidant activity and estimated daily intakes were investigated. There were large variations in the levels of total catechins: 43 and 117 mg g^?1 dry matter (DM) (brewed for 30 s and 5 min respectively); 72 and 161 mg g^?1 DM (extracted in boiling water and 50% acetonitrile respectively); 72 and 117 mg g^?1 DM (a tea bag and tea leaves respectively). The effects on caffeine content were comparatively smaller. These variations consequently led to considerable variations in estimated daily intakes based on three cups (600 ml), ranging between 538 and 2014 mg g^?1 DM of total catechins and between 103 and 466 mg g^?1 DM of caffeine. The antioxidant activity was highest (26 680 µmol g^?1 DM) for tea leaves brewed for 5 min and lowest (10 110 µmol g^?1 DM) for a tea bag product brewed for 1 min. Copyright © 2005 Society of Chemical Industry     

Changes during ensilage in fermentation products,tea catechins,antioxidative activity and in vitro gas production of green tea waste stored with or without dried beet pulp

Wet green tea waste (GTW) was ensiled in laboratory silos with or without dried beet pulp at a ratio of 2:1 on a fresh weight basis. The silos were opened at 15, 30 and 60 days after anaerobic storage; fermentation products, tea catechins, antioxidative activity and in vitro gas production were determined. Acetic acid prevailed over the fermentation and the pH declined to below 4.2 when wet GTW was ensiled alone. Addition of dried beet pulp greatly enhanced lactic acid production and the pH further decreased to below 3.8 with no butyric acid being detected. Approximately 90% and 80% of (?)‐epicatechin gallate and (?)‐epigallocatechin gallate, respectively, were degraded in silage prepared from wet GTW alone, while the degradation was ameliorated in mixed silage to about 55% and 70%, respectively. Antioxidative activity was not markedly altered by ensiling and 70–80% of the initial activity was detected after 60 days. Changes were small in (?)‐epigallocatechin, caffeine and total phenols with or without mixing with dried beet pulp. In vitro gas production was similar between materials and their silages, indicating that reduction of tea catechins would not elicit an improvement in digestibility. It is concluded that wet GTW can be ensiled successfully without bacterial inoculants when mixed with other materials containing certain sugars. Ensiling may degrade part of the components of tea catechins, while not affecting total phenols, antioxidative activity and digestibility of wet GTW. Copyright © 2007 Society of Chemical Industry     

Selective elution of tea catechins and caffeine from polyvinylpolypyrrolidone

Desorption of catechins and caffeine from polyvinylpolypyrrolidone (PVPP) was comprehensively investigated. The result showed that caffeine could easily be desorbed from PVPP by the tested solvents except n‐hexane, while catechins could only be thoroughly done by dimethyl sulphoxide (DMSO) and N, N‐dimethylformamide (DMF). Excellent desorption efficiency of DMSO and DMF might be attributed to their high dipole moments and H‐bond potentials. Addition of ethanol was recommended considering the elution efficiency and fluidity, but ethanol volume should be <40% (v/v) for DMSO or 20% (v/v) for DMF. Desorption would get to equilibrium within 1 h and followed the pseudo‐second‐order model. Caffeine and catechins could be separately desorbed through two‐stage elution procedure, that is, water or 20% aqueous ethanol for desorbing caffeine and part of nongalloylated catechins and DMSO/ethanol (8/2, v/v) for eluting the remaining catechins. Highly purified catechins (~95%) with high level (~70%) galloylated catechins would be achieved when the desorption procedure was applied in column chromatograph.     

基于代谢谱分析的祁门红茶加工过程中儿茶素及芳香类物质变化

祁门红茶是全发酵茶,鲜叶中儿茶素和芳香类物质在加工过程中发生了显著变化。以祁门红茶加工过程中各个阶段的样品为材料,采用高效液相色谱对儿茶素、茶黄素进行定量分析;采用同时蒸馏萃取法提取香气物质,结合气相色谱-质谱联用技术分析香气组分变化,寻找它们在加工中代谢规律。结果表明:儿茶素类在加工过程中一直在降解,烘干叶中儿茶素含量仅为鲜叶8.72%。茶黄素代谢呈现出先增加后减少的趋势,茶黄素总量在揉捻叶中比鲜叶增加了2369.91%,烘干叶比揉捻叶减少了66.44%。茶叶中检测到63种香气组分,其中芳樟醇及其氧化产物、苯乙醇、香叶醇、苯乙醛、苯甲酸、香叶酸和水杨酸甲酯是祁红干茶中主要成分,占香气总量的69.69%,对祁红香气形成起着决定性作用。研究结果对于掌握儿茶素和芳香物质在加工中代谢规律,控制红茶加工过程,提高产品质量都具有一定的理论和实践意义。      

Aldehyde formation in frozen mackerel (Scomber scombrus) in the presence and absence of instant green tea

The effect of frozen storage on lipid peroxidation in Atlantic mackerel (Scomber scombrus) stored for up to 26 weeks at −10 or −80 °C (control), with and without green tea antioxidants, was investigated. Hydroperoxides (PV) and aldehydes (TBARS) were measured by HPLC and LC–MS and hexanal by GC. There was an increase in peroxide value which was associated with an increase in aldehydes, followed by hexanal increase with storage time and at a higher temperature of −10 °C compared with samples stored at −80 °C. Although TBARS is a common assay used to follow malondialdehyde formation, other aldehyde products can also react with thiobarbituric acid to give the red chromogen. Analysis of aldehyde–TBA adducts by LC–MS confirmed the presence of malondialdehyde and, in particular, we report the production of gluteraldehyde for the first time in stored frozen fish. Green tea (at 250 ppm) substantially slowed down the oxidation process, whereas at 500 ppm it was less effective.     

In vitro human skin permeation and cutaneous metabolism of catechins from green tea extract and green tea extract-loaded chitosan microparticles

Catechins are major antioxidants in green tea (Camellia sinensis or Camellia assamica), but because they do not permeate the skin well, the application of green tea in cosmetic products has so far been limited. This study aims to evaluate the cutaneous absorption of catechins from an extract of green tea and from a green tea extract-loaded chitosan microparticle. The catechin skin metabolism was also examined. The results suggest that chitosan microparticles significantly improve the ability of catechins to permeate skin. The cutaneous metabolism of the catechins significantly affected their permeation profiles. Epicatechin (EC) and epigallocatechin (EGC) penetrated the skin more than epigallocatechin gallate (EGCG) and epicatechin gallate (ECG). The galloyl groups in EGCG and ECG were enzymatically hydrolysed to EGC and EC, respectively. Dehydroxylation of catechins was also observed. Chitosan microparticles effectively prevented enzymatic changes of the catechins; therefore, chitosan microparticles are here found to be the promising carriers for enhancing the skin permeation.     

An assessment of dietary supplementation with tea catechins and rosemary extract on the quality of fresh beef

The effect of supplementation of beef cattle diets with tea catechins (TC) (1000mg/animal/day) and rosemary extract (RE) (1000mg/animal/day), for 103 days preceding slaughter, on the oxidative stability of M. longissimus dorsi (LD) steaks was evaluated. Dietary supplementation with TC and RE did not increase plasma total antioxidant status (TAS), LD α-tocopherol concentrations or pH. In LD steaks stored aerobically or in modified atmosphere packs (80% O(2):20% CO(2)) (MAP) for up to 8 days at 4°C, surface redness (CIE 'a' redness value) and lipid stability (TBARS, mg MDA (malondialdehyde)/kg muscle) were not significantly improved as a result of supplementation with TC and RE. Similarly no improvement in the sensory properties and lipid stability of cooked LD slices, stored aerobically or in 30% CO(2):70% N(2) for up to 11 days at 4°C, was observed. An in vitro fermentation study demonstrated that TC and RE were not fermented to any great extent under simulated rumen conditions. Direct addition of TC (1000ppm) and RE (1000ppm) significantly (P<0.05) improved the colour and lipid stability in LD patties stored in 80% O(2):20% CO(2) for up to 8 days at 4°C, thus, demonstrating the antioxidant potential of TC and RE supplements employed in the present study.     

Interactions of digestive enzymes and milk proteins with tea catechins at gastric and intestinal pH

The interactions of digestive enzymes (pepsin, pancreatin) and milk proteins (β‐casein, β‐lactoglobulin (β‐Lg)) with (?)‐epigallocatechin gallate (EGCG), (?)‐epigallocatechin (EGC) and (?)‐epicatechin (EC) at gastric and intestinal pH were investigated by fluorescence spectroscopy. The results indicated that in the gastric environment, all three tea catechins showed binding affinities in descending order of strength with β‐casein first, followed by β‐Lg and then pepsin. The highest affinity was observed for EGCG–β‐casein, with a binding constant (K_A) of 2.502(±0.201) × 10⁵ m ^?1. In the intestinal environment, the binding strengths of the proteins with EGCG and EGC were in the order β‐Lg > pancreatin > β‐casein; for binding with EC, the strength order was β‐casein > β‐Lg > pancreatin. The combination EGCG–β‐Lg had the strongest binding affinity, with a K_A of 14.300(±0.997) × 10⁵ m ^?1. Thermodynamic analysis revealed that tea catechins complexed with milk proteins and digestive enzymes via different hydrophilic and hydrophobic interactions depending on the different digestion environments and types of catechins, proteins and enzymes.     

The chemical oxidation of catechins and other phenolics: A study of the formation of black tea pigments

The chemical oxidation of polyphenols has been used to produce black tea pigments. (+)-Catechin, (?)-epicatechin, (?)-epicatechin gallate and (?)-epigallocatechin gallate were isolated from convenient sources, their identity being confirmed by mass spectrometry and HPLC. The oxidation of a mixture of (?)-epicatechin gallate and gallic acid gave an oxidation product of (?)-epicatechin gallate in addition to epitheaflavic acid 3-gallate. The catechins were oxidised chemically and the products analysed by HPLC, the chromatograms being compared with a black tea chromatogram. The reactions gave both resolved and unresolved pigments, and many of the resolved pigments had HPLC retention times close to black tea pigments. Each catechin behaved differently, the chromatograms of the oxidation products from each starting material being clearly distinguishable. Resolved pigments were obtained by the chemical oxidation of other phenolic compounds. This work provides a convenient method for studying the formation of resolved and unresolved black tea pigments.     

Tea extraction methods in relation to control of epimerization of tea catechins

Effect of water temperature and ethanol concentration on epimerization and extractability of tea catechins was investigated. The results showed that epigallocatechin gallate (EGCG) and epicatechin gallate (ECG) were partially epimerized into gallocatechin gallate (GCG) and catechin gallate (CG), respectively, when tea catechins extract was heated in water solution at 100 °C for 2 h or dry tea was extracted in water at 100 °C. The epimerization of the catechins was inhibited if the tea catechins extract was heated as solid powder and the dry tea was extracted in 50% (v/v) ethanol or in water at 80 °C or below. When the dry tea was extracted in water, the extractability of catechins increased with the increase of extraction temperature up to 100 °C, but there was no statistically significant difference in total catechins between 80 °C and 100 °C. When teas were extracted using ethanol solutions, the highest extractability of total catechins was obtained in 50% (v/v) ethanol for dry tea and in 75% (v/v) ethanol for fresh tea leaf. In order to reveal the real profiles of tea catechins in teas to be tested, dry tea should be extracted in 50% (v/v) ethanol for 10 min, while fresh tea leaf should be extracted in 75% (v/v) ethanol for 10 min. For commercial extraction, temperature should be controlled at 80 °C if water is used as the solvent. Copyright © 2007 Society of Chemical Industry     

Anti-oxidant activity of added tea catechins on lipid oxidation of raw minced red meat, poultry and fish muscle

The comparative anti-oxidative effects of added tea catechins (TC) and α-tocopherol to raw minced red meat (beef and pork), poultry (chicken, duck and ostrich) and fish (whiting and mackerel) muscle on susceptibility to lipid oxidation were investigated during 10 days of refrigerated (4 °C) display. Fresh meats, poultry and fish, purchased from a local market, were trimmed to remove bones, skin and surface fat and minced through a 4 mm plate. The minced muscle of each species was treated with either the addition of 300 mg TC kg^–1 minced muscle (TC300) or 300 mg α-tocopherol kg^–1 minced muscle (VE300). Minced muscle without any additives served as control (C). Oxidative stability (TBARS) was measured at 3-day intervals. Total lipids, fatty acid composition, total iron and haem iron from minced muscle for each species were also analysed. The susceptibility of untreated minced muscle to lipid oxidation was in the decreasing order: mackerel > beef > duck > ostrich > pork ≥ chicken > whiting. This may be because of the different content of total fat, iron and fatty acid composition between species. The TC300 significantly ( P  < 0.05) reduced lipid oxidation compared with controls for all seven species as shown by lower TBARS values. The anti-oxidant potential of TC was two to fourfold greater than that of α-tocopherol at the same concentration and this potential was species dependent. The VE300 showed limited capacity in inhibiting lipid oxidation for pork, chicken, duck and whiting. The results obtained show that TCs are powerful natural antioxidants when used in minced muscle food.     

Stability of catalase and its potential role in lipid oxidation in meat

The activity of catalase in microbial growth-controlled and uncontrolled ground beef muscle (semimembranosus, SM) did not change (P>0.05) during 6-day storage at 4°C. Likewise, catalase activity in ground, beef SM and longissimus dorsi (LD), pork LD, and chicken breast (B) and thigh (T) muscles was not affected (P>0.05) by 2-month storage at −20°C, with or without mid-month thawing/refreezing. When sodium azide (a catalase inhibitor) was added to ground beef SM, lipid oxidation (as measured by peroxide values) during 4-day refrigeration was higher (P<0.05) in treated samples — 43 and 55% higher at day 2 and day 4, respectively — than in the controls. It was concluded that catalase would be stable during meat storage/distribution and contribute significantly to the antioxidative process in raw meat products.