Measurement of 2-thiobarbituric acid values in dark chicken meat through derivative spectrophotometry: influence of various parameters

Several variables (kind of filter paper, amount of sample, antioxidant addition, stability of the spectrophotometric measurement, and handling and storage of samples) were found to influence 2-thiobarbituric acid (TBA) values in dark chicken meat when an acid aqueous extraction method with derivative spectrophotometry was used. Filter papers with larger pore diameter or increasing sample weight led to lower TBA values. After incubation of reaction mixtures at 70 degrees C and ice-cooling, tempering for 45 min at room temperature was necessary to stabilize the spectrophotometric measurement. Furthermore, addition of butylated hydroxytoluene (BHT) and ethylenediaminetetraacetic acid disodium salt (EDTA) in the early steps of the method prevented artifactual formation of TBA reactive substances during analysis. Vacuum packaging and storage of samples at -20 degrees C were useful to avoid sample oxidation. The method finally proposed has a coefficient of variation of 3.81 or 4.13% for raw or cooked samples, respectively.     

Using a modified ferrous oxidation-xylenol orange (FOX) assay for detection of lipid hydroperoxides in plant tissue.

The ferrous oxidation-xylenol orange (FOX) assay was adapted for quantifying lipid hydroperoxides (LOOHs) in plant extracts. Excised pieces of several fruit and vegetable species were exposed to 83 kJ m(-2) day(-1) of biologically effective ultraviolet B irradiance (UV-B(BE)) for 10-12 days to induce cellular oxidation. The LOOH and thiobarbituric acid reactive substance (TBARS) concentrations of these plant tissues were assessed with the FOX and iodometric assays for the former and a modified TBARS assay for the latter. There was generally good agreement between the FOX and iodometric methods both prior to and following the UV exposure. However, the iodometric assay appeared to have some difficulty in consistently quantifying lower LOOH levels (<11 microM), whereas the FOX assay measured LOOH concentrations as low as 5 microM. All tissues exhibited UV-induced increases in TBARS, indicating a marked degree of cellular oxidation in the exposed tissue segments. Compared with the iodometric assay, the FOX method consistently generated less variable LOOH values. The presence of authentic linoleic acid-OOHs in spiked avocado and spinach samples (11 microM) was identified with liquid chromatography-mass spectrometry techniques, which validated corresponding FOX assay results. The FOX method is inexpensive, is not sensitive to ambient O2 or light levels, and can rapidly generate LOOH measurements. The physiological value of the FOX assay resides in its ability to measure initial rather than more advanced fatty acid oxidation; hence, early membrane-associated stress events in plant tissue can be detected.     

Determination of lipoxygenase activity in plant extracts using a modified ferrous oxidation-xylenol orange assay

The ferrous oxidation-xylenol orange (FOX) assay method for determination of lipid hydroperoxides is based on that under acidic conditions Fe2+ is oxidized to Fe3+, which then oxidizes xylenol orange to a product that absorb at 550 nm. The procedure has been adapted for determination of lipoxygenase activity in plant extracts. This enzyme is responsible for generation of off-flavors in vegetal foods, bleaching of pigments, and a lot of oxidative degradations. It is of interest to check the initial lipoxygenase activity in vegetal foods before the processing, using an assay that is rapid, reproducible, and easily adaptable to high throughput format. The enzymatic assay is based on a discontinuous determination of lipoxygenase activity using the FOX reagent for colorimetric determination of hydroperoxides accumulated in the medium by a period of incubation that is established by the addition of the extract (start of the reaction) and the addition of FOX reagent (finish of the reaction). The procedure is capable of detecting lipoxygenase activity in a number of vegetable homogenates, being especially useful for a rapid visual evaluation of this enzymatic activity.     

Antioxidative activity and antidiscoloration efficacy of ergothioneine in mushroom (Flammulina velutipes) extract added to beef and fish meats

The antioxidative property of a hydrophilic extract prepared from the fruiting body of edible mushroom ( Flammulina velutipes) was evaluated. The mushroom extract contained ergothioneine (ERT) at a level of 3.03 +/- 0.07 mg/mL, showed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and suppressed lipid oxidation of bigeye tuna meat more effectively than authentic L-ERT added at the same concentration. The authentic L-ERT had stronger total reducing power than the mushroom extract and inhibited the formation of metmyoglobin (metMb) more significantly in bigeye tuna meat. Lipid oxidation in beef and fish meats to which the mushroom extract had been added was "virtually" controlled during storage on ice. Ground beef and bigeye tuna meat with the extract added kept their natural colors unchanged for longer than 12 and 7 days of ice storage, respectively. Contrary to this, browning in meat color was observed in the control samples without the extract after 6 and 2 days of storage, respectively, when stored under similar conditions. There was significant correlation between meat color and chemical parameters, including total lipid hydroperoxides, thiobarbituric acid reactive substances, and metMb. However, there was no significant correlation between pH value and meat discoloration. These results suggest that ERT in the hydrophilic extract of F. velutipes plays an important role as a color stabilizer of meats.     

Automated dynamic headspace/GC-MS analyses affect the repeatability of volatiles in irradiated Turkey

Although a dynamic headspace/gas chromatography-mass spectrometry (DH/GC-MS) method is an effective tool for determining volatiles of irradiated turkey meat, the profile of volatiles may be changeable depending upon the availability of oxygen in the sample vial and sample holding time before purge. The objective of this study was to evaluate the effects of helium flushing and sample holding time before purge on the volatiles profiles of irradiated raw and cooked turkey breast meat. Vacuum-packaged turkey breasts were irradiated at 2.5 kGy, and the volatiles of irradiated raw and cooked samples were analyzed using a DH/GC-MS with different holding times up to 280 min. The amounts of dimethyl disulfide and dimethyl trisulfide decreased as sample holding time in an autosampler (4 degrees C) before purge increased, whereas those of aldehdyes increased as holding time increased due to lipid oxidation. Helium flush of sample vials before sample loading on an autosampler retarded lipid oxidation and minimized the changes of sulfur volatiles in raw meat but was not enough to prevent oxidative changes in cooked meat. Although DH/GC-MS is a convenient method for automatic analysis of volatiles in meat samples, the number of samples that can be loaded in an autosampler at a time should be limited within the range that can permit reasonable repeatabilities for target volatile compounds.     

柑橘幼果提取物对猪肉冷藏过程中抗脂质氧化影响

为了研究不同浓度的柑橘幼果乙醇提取物（CE）在新鲜猪肉中的抗脂质氧化和抑菌作用。在冷藏（4℃）条件下对鲜猪肉进行以下5种处理：对照组（不添加保鲜剂）;AC-0.02（添加0.02%抗坏血酸）;CE-0.05（添加0.05%柑橘幼果醇提物,下同）,CE-0.1和CE-0.2。在12 d的冷藏（4℃）过程中,各样品的pH值均显著降低（p＜0.05）,而硫代巴比妥酸值（TBARS）和游离脂肪酸的含量显著增加（p＜0.05）。CE-0.1和CE-0.2处理组的菌落总数在贮藏过程中均低于对照组。CE的加入使猪肉样品的L*值和a*值都显著下降,在贮藏过程中,CE处理组样品的b*值均高于对照组,且随着CE浓度的增加,b*值增大。5组试验样品中,经CE-0.2处理的样品TBARS值和游离脂肪酸含量最低;对照组的过氧化值在第6天达到最大值,随后逐渐减小,而其它处理组的过氧化值在储藏期间均呈上升趋势。在贮藏1 d后,各处理组中的共轭二烯值均小于对照组。结果表明,柑橘幼果醇提物可作为肉制品的保鲜剂,能有效抑制新鲜猪肉贮藏过程中的脂质氧化和腐败微生物的生长。为开发柑橘幼果天然提取物保鲜剂提供理论参考。     

Far-infrared radiation increases the antioxidant properties of rice hull extract in cooked turkey meat

To determine the antioxidant effects of rice hull extract exposed to far-infrared radiation, the added extracts were compared with sesamol in cooked turkey breast. Rice hull extract showed antioxidant properties in cooked turkey breast by reducing lipid oxidation and volatile aldehydes. Far-infrared radiation increased significantly the antioxidant activities of rice hull extracts. Rice hull extract irradiated by far-infrared (FRH) had lower TBARS values and fewer volatile aldehydes (hexanal, pentanal, and propanal) than a non-irradiated extract (IRH) during the 3 days of aerobic storage. Addition of FRH at 0.2% (w/w) in turkey meat could reduce the amounts of volatile hexanal to 18-47% of the control during the storage. However, the antioxidant activities of rice hull extracts did not last as long as those of pure sesamol due to the relatively low concentration of phenolics, and the extracts had some peculiar odor. Addition of rice hull extracts also increased both a and b values of the samples due to its brown intensity.     

Sensitive Quantification of Silver Nanoparticles by Kinetic-Spectrophotometry Method in Groundwater Samples

A novel kinetic-spectrophotometry method is developed for the determination of trace amounts of silver nanoparticles (AgNPs) in groundwater samples based on their catalytic effect on the oxidation of indigo carmine by hexacyanoferrate (III) in acetate-acetic acid medium. The absorbance was measured at 612?nm with the fixed-time method. The optimization of the operating conditions regarding concentrations of the reagents and interferences was investigated. The calibration curve is linear over the concentration range 4.0?C300???g?mL^?1 of AgNPs with good precision and accuracy. The proposed method is a highly sensitive, selective, and relatively rapid assay of AgNPs at trace level. The method was applied to the determination of AgNPs in groundwater and a synthetic sample.     

Quantitative analysis and detection of adulteration in crab meat using visible and near-infrared spectroscopy

Visible and near-infrared spectroscopy (VIS/NIR) has been used to detect economic adulteration of crab meat samples. Atlantic blue and blue swimmer crab meat samples were adulterated with surimi-based imitation crab meat in 10% increments. Waveform evaluation revealed that the main features seen in the spectral data arise from water absorptions with a decrease in sample absorbance with increasing adulteration level. Prediction and quantitative analysis was done using raw data, a 15-point smoothing average, a first derivative, a second derivative, and 150 wavelength spectral data gathered from a correlogram. Regression analysis included partial least squares (PLS) and principal component analysis (PCR). Both models were able to perform similarly in predicting crab meat adulteration. The best model for both PLS and PCR used the first derivative spectral data gathered from the correlogram, with a standard error of prediction (SEP) of 0.252 and 0.244, respectively. The results suggest that VIS/NIR technology can be successfully used to detect adulteration in crab meat samples adulterated with surimi-based imitation crab meat.     

Thiol-quinone adduct formation in myofibrillar proteins detected by LC-MS

Protein oxidation in meat is considered to decrease meat tenderness due to protein disulfide cross-link formation of thiol-containing amino acid residues. An LC-MS method for detection of thiol-quinone adducts (RS-QH(2)) in myofibrillar proteins was developed to investigate the interaction between phenols, as protective antioxidants, and proteins from meat under oxidative conditions using aqueous solutions of (i) cysteine (Cys), (ii) glutathione (GSH), (iii) bovine serum albumin (BSA), or (iv) a myofibrillar protein isolate (MPI). The aqueous solutions were incubated at room temperature (30 min) with 4-methyl-1,2-benzoquinone (4MBQ) prepared from oxidation of 4-methylcatechol (4MC) by periodate resin or incubated at room temperature (5 h) with 4MC and Fe(II)/H(2)O(2). GSH, BSA, and MPI were hydrolyzed (6 N HCl, 110 °C, 22 h) after incubation, and the cysteine-quinone adduct, Cys-QH(2) (m/z 244.2) was identified according to UV and mass spectra after separation on an RP-C18 column. The thiol-quinone adduct was present in all thiol systems after incubation with 4MBQ or 4MC oxidized by Fe(II)/H(2)O(2). Direct reaction with 4MBQ resulted in each case in increased Cys-QH(2) formation compared to simultaneous oxidation of thiol source and 4MC with Fe(II)/H(2)O(2). The covalent bonds between quinones and thiol groups may act as a potential antioxidant by inhibiting disulfide protein cross-link formation.     

Colorimetric determination of hydroxyproline as measure of collagen content in meat and meat products: NMKL collaborative study

A colorimetric method for the determination of hydroxyproline as a measure of collagen in meat and meat products has been collaboratively studied in 18 laboratories. The method includes hydrolysis with sulfuric acid, oxidation with chloramine-T, and formation of a reddish purple complex with 4-dimethylaminobenzaldehyde. Five frozen and 3 freeze-dried samples were tested, ranging in content from 0.11 to 0.88% and from 0.39 to 4.0% hydroxyproline, respectively. The mean values of 2 identical samples were 0.245 and 0.251%. The average recovery from a spiked sample was 96.1%. The hydroxyproline content of a known sample (a mixture of 2 samples in the ratio 5:2) was calculated to 1.42%, which agrees well with the analytical result, 1.40%. In comparison with other collaborative studies, based on the ISO analytical method, the repeatability and reproducibility of this method agree well with the other results. This method was accepted as an official NMKL method by all national Committees, and has been adopted official first action by AOAC as an NMKL-AOAC method.     

“吸光量”的原理、技术和应用

本文概述了吸光量的概念、原理、测试技术和一些应用实例。     

Development of a sensitive and rapid method for the measurement of total microbial activity using fluorescein diacetate (FDA) in a range of soils

Fluorescein diacetate (FDA) hydrolysis is widely accepted as an accurate and simple method for measuring total microbial activity in a range of environmental samples, including soils. Colourless fluorescein diacetate is hydrolysed by both free and membrane bound enzymes, releasing a coloured end product fluorescein which can be measured by spectrophotometry. The current method for measuring FDA hydrolysis in soils is limited in its application. FDA activity was very low in sandy and clayey soils. The low activity observed for these soil types was made difficult to measure by the original authors’ choice of solvent for terminating the hydrolysis reaction. Acetone (50% v/v) was found to be most efficient at stopping the hydrolysis reaction. During this study acetone (50% v/v) was found to cause a decrease of approximately 37% in the absorbance of fluorescein produced by the soil samples measured. Although this colour loss is independent of initial fluorescein concentration, it makes the measurement of FDA hydrolytic activity extremely difficult in soils with low microbial activity i.e. sandy and/or clayey soils. Chloroform/methanol (2:1 v/v) was found to successfully stop the hydrolysis reaction for up to 50 min in a range of soil samples without causing the loss of colour observed with acetone. By changing the solvent used for terminating the hydrolysis reaction, low activity soils could be measured successfully. Other parameters of the hydrolysis reaction were optimised for the measurement of soil samples including effect of pH, optimum temperature of incubation, amount of soil, time of incubation, amount of substrate and preparation of suitable standards. A new, more sensitive method is proposed adapted from the original method, which provides a more accurate determination of FDA hydrolysis in a wide range of soils.     

Isomeric influence on the oxidative coloration of phenolic compounds in a model white wine: comparison of (+)-catechin and (-)-epicatechin

The reactions of (+)-catechin and (-)-epicatechin with glyoxylic acid were studied in a model white wine solution. When the reactions were performed in darkness at 45 degrees C, the (-)-epicatechin concentration decreased more rapidly than that of (+)-catechin, and the (-)-epicatechin sample had twice the 440 nm absorbance of the (+)-catechin sample after the 14 day incubation period. The main pigments generated were identified as xanthylium cation pigments regardless of the isomeric character of the phenolic compound. Using a combination of absorbance and ion current data, the xanthylium cation pigments generated from (-)-epicatechin were found to have combined molar absorptivity coefficients 1.8 times that of the xanthylium cation pigments generated from (+)-catechin. The implication of these results on the development of an index of white wine oxidation susceptibility is discussed.     

Potato peel extract-a natural antioxidant for retarding lipid peroxidation in radiation processed lamb meat

The effective utilization of potato peel, a waste generated in large quantities by the food industry, as an antioxidant was investigated. Potato peel extract (PPE) exhibited high phenolic content (70.82 mg of catechin equivalent/100 g), chlorogenic acid (27.56 mg/100 g of sample) being the major component. The yield of total phenolics and chlorogenic acid increased by 26 and 60%, respectively, when the extract was prepared from gamma irradiated (150 Gy) potatoes. PPE showed excellent antioxidant activity as determined by beta-carotene bleaching and radical scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH). The suitability of PPE for controlling lipid oxidation of radiation processed lamb meat was also investigated. PPE (0.04%) when added to meat before radiation processing was found to retard lipid peroxidation of irradiated meat as measured by TBA number and carbonyl content. The antioxidant activity of PPE was found to be comparable to butylated hydroxytoluene (BHT).     

辐照处理提高猪肉火腿肠保鲜效果

辐照是一种非常有效的食品杀菌保鲜技术,近年来在即食肉制品保鲜方面的应用逐渐引起了人们的关注。为了探讨γ辐照技术对猪肉火腿肠的杀菌保鲜效果,试验采用1、3、5、7、9 k Gy 5个剂量分别对火腿肠进行辐照处理,辐照结束后样品在4℃条件下冷藏,然后跟踪测定其在冷藏过程中脂肪和蛋白质氧化、颜色、p H值、挥发性盐基氮(total volatile basic nitrogen,TVB-N)以及菌落总数的变化,分析不同剂量辐照处理对火腿肠保鲜效果的影响。结果表明,辐照可以显著降低火腿肠脂肪和蛋白质的氧化稳定性,促进其在储藏过程中的氧化。1 k Gy剂量辐照对火腿肠的色泽影响不显著;3~9 k Gy剂量辐照,可以使样品的红度值(a*值)显著降低、亮度值(L*值)显著升高(P0.05),而在冷藏过程中a*值与L*值均随储藏时间的延长而降低,且高剂量辐照组要比低剂量组降低更快一些。火腿肠p H值受辐照影响不显著(P0.05),且在冷藏过程中所有处理组火腿肠的p H值变化也不显著,均在6.0~6.5范围内。5 k Gy剂量辐照即可对猪肉火腿肠中的微生物起到有效抑制作用。     

扩散法测定土壤无机氮~(15)N丰度方法优化研究

扩散法与质谱测定技术相结合,被广泛应用于环境、生态和农业领域中土壤、水等样品中无机氮15N同位素丰度的测定。为建立一套可快速、准确测定土壤无机氮15N同位素丰度的扩散培养体系,针对土壤样品无机氮量的变化特点,从培养温度、培养时间、试剂选择和用量等方面对扩散条件进行优化。结果发现,对于大部分无机氮浓度大于2 mg L~(-1)的土壤样品,20 ml土壤提取液,在不小于250 ml的蓝盖瓶中,悬挂两张各滴加了10μl 1 mol L~(-1)草酸的滤纸,加入0.1 g的Mg O,25℃下以140 r min-1的转速振荡培养24 h即可完成对样品中NH4+-N的扩散与回收;换入2张加酸滤纸继续摇培48 h可基本去除残余的NH4+-N;再换入2张加酸滤纸并加入0.1 g的戴氏合金振荡培养24 h即可。对于无机氮浓度低于2 mg L~(-1)的土壤提取液,需用50 ml提取液按以上条件进行扩散培养即可保证测定结果的准确性。本方法大大缩短了扩散法所需的实验周期,实现在一份样品内同时完成NH4+-N和NO3--N的扩散与回收,减少了样品的需要量,并通过优化Mg O、戴氏合金(Devarda’s alloy)的用量减少杂质氮可能带来的污染。     

电子鼻和电子舌单独与联合检测掺大豆蛋白或淀粉的鸡肉糜

为实现掺杂掺假鸡肉的快速、客观评价,该研究利用电子鼻和电子舌联合检测技术对掺杂鸡肉糜进行快速检测,通过对采集的数据进行主成分分析和偏最小二乘法分析,所得结果表明,采用主成分分析,电子鼻和电子舌联合检测掺大豆蛋白鸡肉糜和掺淀粉鸡肉糜的主成分总贡献率分别为99.8%和99.1%;采用偏最小二乘法分析,电子鼻和电子舌联合检测鸡肉糜中掺杂大豆蛋白含量的预测值与真实值之间的决定系数为0.992,均方根误差为2.8%;联合检测鸡肉糜中掺杂淀粉含量的预测值与真实值之间的决定系数为0.996,均方根误差为2.4%。表明电子鼻和电子舌联合检测对鸡肉糜的掺杂情况具有良好的区分和预测能力,并且是一种有效、高精度的肉类掺假检测方法。     

Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method

The chemical diversity of antioxidants makes it difficult to separate and quantify antioxidants from the vegetable matrix. Therefore, it is desirable to establish a method that can measure the total antioxidant activity level directly from vegetable extracts. The current literature clearly states that there is no "total antioxidant" as a nutritional index available for food labeling because of the lack of standard quantitation methods. Thus, this work reports the development of a simple, widely applicable antioxidant capacity index for dietary polyphenols and vitamins C and E, utilizing the copper(II)-neocuproine [Cu(II)-Nc] reagent as the chromogenic oxidizing agent. Because the copper(II) (or cupric) ion reducing ability of polyphenols is measured, the method is named by our research group "cupric reducing antioxidant capacity" abbreviated as the CUPRAC method. This method should be advantageous over the ferric reducing antioxidant power (FRAP) method because the redox chemistry of copper(II)-as opposed to that of ferric ion-involves faster kinetics. The method comprises mixing of the antioxidant solution (directly or after acid hydrolysis) with a copper(II) chloride solution, a neocuproine alcoholic solution, and an ammonium acetate aqueous buffer at pH 7 and subsequent measurement of the developed absorbance at 450 nm after 30 min. Because the color development is fast for compounds such as ascorbic acid, gallic acid, and quercetin but slow for naringin and naringenin, the latter compounds were assayed after incubation at 50 degrees C on a water bath for 20 min [after Cu(II)-Nc reagent addition] so as to force the oxidation reaction to reach completion. The flavonoid glycosides were hydrolyzed to their corresponding aglycons by refluxing in 1.2 M HCl-containing 50% MeOH so as to exert maximal reducing power toward Cu(II)-Nc. Certain compounds also needed incubation after acid hydrolysis to fully exhibit their reducing capability. The CUPRAC antioxidant capacities of synthetic mixtures of antioxidants were experimentally measured as Trolox equivalents and compared to those theoretically found by making use of the principle of additivity of absorbances assuming no chemical interaction between the mixture constituents. Because ascorbic acid is not resistant to elevated temperature incubation, it should be assayed initially by measuring the absorbance (at 450 nm) difference of original and ascorbate oxidase-added mixture solutions at the end of 1 min of Cu(II)-Nc reagent addition. Thus, the total CUPRAC antioxidant capacity of a mixture containing various antioxidants should be that finally measured after a suitable combination of hydrolysis and incubation procedures, added to the initially measured capacity due to ascorbate. The antioxidant polyphenolic compounds tested demonstrate that the highest capacities in the CUPRAC method were observed for epicatechin gallate, epigallocatechin gallate, quercetin, fisetin, epigallocatechin, catechin, and caffeic acid in this order, in accordance with theoretical expectations, because the number and position of the hydroxyl groups as well as the degree of conjugation of the whole molecule are important. The antioxidant potency of flavonoids is nearly proportional to the total number of -OH groups and is positively affected by the presence of an o-dihydroxy moiety in the B-ring. beta-Carotene, which did not react with the CUPRAC reagent in alcoholic aqueous medium, could be assayed in dichloromethane solvent. Linear calibration curves for ascorbic acid and flavonoids were redrawn in synthetic solutions containing a mixture of antioxidants, and also in real matrices such as grape and orange juices, green tea, and blackberry tea, showing an initial nonzero absorbance with the CUPRAC reagent. The parallellism of the linear calibration curves of pure compounds in a given complex matrix effectively demonstrated that there were no interferent chemical interactions among the solution constituents and that the antioxidant capacities of the tested antioxidants were additive. The CUPRAC reagent is reasonably selective, stable, easily accessible, and sensitive toward thiol-type oxidants, unlike the FRAP method. The reaction is carried out at nearly physiological pH as opposed to the unrealistic acidic pH of FRAP.