基于海藻酸钠与鸡蛋黄静电聚集作用的低脂蛋黄酱制备

为了拓展海藻酸钠和鸡蛋黄蛋白质在低脂蛋黄酱质构设计方面的应用,该研究首先探究了海藻酸钠和鸡蛋黄分散液在不同酸性pH值条件下的聚集行为,并基于两者的静电聚集作用设计出油相比为30%(体积分数)且具有明显黏弹性和触变性的低脂蛋黄酱产品,同时以油相比为75%的蛋黄酱作对照。结果表明,当pH值低于5.0时,海藻酸钠携带负电荷,鸡蛋黄分散液携带正电荷,两者可发生明显的静电聚集作用,海藻酸钠和鸡蛋黄复合体系的结构强度增加。当白醋添加量高于2%(体积分数)时,海藻酸钠和鸡蛋黄复合体系的pH值降低至5.0以下,可诱导复合体系发生静电聚集作用,白醋添加量越高,聚集作用越明显,低脂蛋黄酱的结构化程度也越高。然而,过量的白醋添加降低了低脂蛋黄酱的热稳定性,同时也影响了产品的风味和感官接受度。综合而言,当白醋添加量为4%时(pH值4.6),制备的低脂蛋黄酱流变学特性和对照组最为接近,且感官接受度较好。该研究结果可为构建低脂食品提供理论参考。     

Determination of advanced glycation endproducts by LC-MS/MS in raw and roasted almonds (Prunus dulcis)

A sensitive and reliable LC-(ESI)MS/MS method was developed and validated for the simultaneous analysis of five common advanced glycation endproducts (AGEs) after enzymatic digestion in raw and roasted almonds. AGEs included carboxymethyl-lysine (CML), carboxyethyl-lysine (CEL), pyralline (Pyr), argpyrimidine (Arg-p), and pentosidine (Pento-s). This method allows accurate quantitation of free and AGE-protein adducts of target AGEs. Results indicate that CML and CEL are found in both raw and roasted almonds. Pyr was identified for the first time in roasted almonds and accounted for 64.4% of free plus bound measured AGEs. Arg-p and Pento-s were below the limit of detection in all almond samples tested. Free AGEs accounted for 1.3-26.8% of free plus bound measured AGEs, indicating that protein-bound forms predominate. The roasting process significantly increased CML, CEL, and Pyr formation, but no significant correlation was observed between these AGEs and roasting temperature.     

咸蛋黄快速腌制过程中理化性质变化规律

该研究以新鲜蛋黄为原料,利用快速腌制模具,探究在咸蛋黄的上表面添加食盐单侧腌制过程中,食盐添加量和腌制时间对咸鸡蛋黄快速腌制过程中形貌特征和理化性质变化规律的影响。借助多种仪器分析手段对蛋黄腌制过程中形貌与物性的变化、水分及盐分的迁移规律进行了表征。低场核磁及成像结果表明：在腌制过程中,蛋黄中的水分不断向外迁移,含水率显著降低,当增加食盐的添加量和延长腌制时间,会加快水分的迁移速率;原子吸收结果表明：增加食盐添加盐量越多和腌制时间越长盐分迁移速率越快,质构、色差结果共同表明咸蛋黄腌制过程中,由于水分的向外迁移和盐分的向内渗入,使得蛋黄的蛋白质发生聚集使颜色加深;同时与市售整个腌制后分离的鸡蛋黄产品相比,当腌制时间为7d,添加盐量为3%;腌制时间为3d,添加盐量为5%时,所得的样品与市面的成品咸鸡蛋黄的感官品质及量化指标差异不显著（P<0.05）,为咸蛋黄单侧腌制技术提供理论的可行性。     

Conjugated linoleic acids alter the fatty acid composition and physical properties of egg yolk and albumen

Effects of dietary conjugated linoleic acids (CLAs) and docosahexaenoic acid (DHA) on the fatty acid composition of different egg compartments after storage were studied. Four dietary treatments [supplemented with safflower oil (SAFF, control group), DHA, CLAs plus DHA (CAD), and CLAs alone] were administered to Single Comb White Leghorn (SCWL) laying hens. Eggs from the different treatment groups were collected and stored for 10 weeks at 4 degrees C before analysis. Fatty acids from the yolk (yolk granules and plasma), egg albumen, and vitelline membrane were analyzed by gas chromatography. The yolk of eggs from hens given CLAs had significantly higher amounts of saturated fatty acids, typically 16:0 and 18:0, but lower amounts of polyunsaturated fatty acids (PUFAs) compared to eggs from the control group (SAFF). CLA content was highest in the yolk and present in both neutral and polar lipids, with the greatest concentrations in neutral lipids. DHA was incorporated mainly into yolk polar lipids. Lipids in yolk plasma and granules contained similar amounts of CLAs. The fatty acid compositions of vitelline membrane and egg albumen mirrored that of the egg yolk. CLA supplementation resulted in hard and rubbery yolks when compared to hard-cooked eggs from the control group. This study showed that feeding CLAs to hens led to accumulation of the isomers in polar and neutral lipids of the egg yolk and that these isomers migrated into egg albumen. Because the sensory properties of hard-cooked eggs were negatively affected by the enrichment of a mixture of CLA isomers in this study, further research should be conducted to evaluate how the different isomers alter the properties of egg yolk and albumen so that the quality of designed eggs containing CLAs and DHA can be improved.     

Residues of veterinary drugs in eggs and their distribution between yolk and white

Veterinary drugs and feed additives (especially some coccidiostats) can be absorbed by the digestive tract of laying hens and transferred to the egg. Physicochemical characteristics of these compounds determine their pharmacokinetic behavior and distribution to and within the egg. Traditionally the quite lipid soluble drugs and additives are expected to yield residues only in the fat-rich yolk. However, the quite lipid soluble drug doxycycline--as well as many other drugs--showed during long-term administration higher residues in white than in yolk. In a model study with 11 sulfonamides differing in pK(a) value and lipid solubility, their distribution in vivo between yolk and white was determined. Neither differences in pK(a) values nor those in lipid solubility could explain the distributions found. Binding to egg white macromolecules in vivo as an explanatory factor was tested with five sulfonamides, and no correlation between binding and the distribution of sulfonamides between white and yolk was found. Literature data on the distribution of drugs between egg white and yolk showed a reasonable consistency within drugs and a large variability among drugs (as could be expected). This larger database also did not provide a clue as to what factor determines the distribution of a drug between egg white and yolk when given to laying hens.     

Development of yellow pigmentation in squid (Loligo peali) as a result of lipid oxidation

The impact of lipid oxidation on yellow pigment formation in squid lipids and proteins was studied. When the squid microsomes were oxidized with iron and ascorbate, thiobarbituric acid reactive substance were observed to increase simultaneously with b values (yellowness) and pyrrole compounds concomitantly with a decrease in free amines. Oxidized microsomes were not able to change the solubility, sulfhydryl content, or color of salt-soluble squid myofibrillar proteins. Aldehydic lipid oxidation products were able to decrease solubility and sulfhydryl content of salt-soluble squid myofibrillar proteins but had no impact on color. Aldehydic lipid oxidation products increased b values (yellowness) and pyrrole compounds and decreased free amines in both squid phospholipid and egg yolk lecithin liposomes. The ability of aldehydic lipid oxidation products to change the physical and chemical properties of egg yolk lecithin liposomes increased with increasing level of unsaturation and when the carbon number was increased from 6 to 7. These data suggest that off-color formation in squid muscle could be due to nonenzymatic browning reactions occurring between aldehydic lipid oxidation products and the amines on phospholipids headgroups.     

New insights into the structure of apolipoprotein B from low-density lipoproteins and identification of a novel YGP-like protein in hen egg yolk

Apoproteins of low-density lipoproteins (LDL) and soluble proteins (livetins) contained in hen egg yolk plasma have been demonstrated as being essential to the interfacial and emulsifying properties of yolk. The knowledge of their structure is necessary to better understand these properties. Purified protein fractions were separated by SDS-PAGE or 2D-PAGE and identified through the LC-MS/MS of their trypsin peptides. Hen blood apolipoprotein B gives rise to nine different apoproteins in LDL after maturation and proteolysis. Among these apoproteins, two protein fragments appeared to be less accessible to proteases and could be enriched in beta-sheets and firmly associated with lipids. Plasma soluble proteins were constituted by approximately 45% of yolk immunoglobulins with a high heterogeneity of the variable regions of both heavy and light chains, 41% of glycoproteins constituted by YGP42 and YGP40, 14% of albumins, and one new minor protein we called YGP30, showing 75% similarity to YGP40.     

Differential incorporation of conjugated linoleic acid isomers into egg yolk lipids

The incorporation pattern of conjugated linoleic acids (CLA) isomers into the egg yolk of hens in relation to that in the diet was studied. Silver-ion high-performance liquid chromatography (Ag-HPLC) was used to separate individual CLA isomers. It was found that the isomeric distribution pattern in the egg yolk lipids was different from that in the dietary fat. Total cis/trans isomers accounted for 81.2% of total CLA incorporated into the egg yolk, which was in contrast to the value of 92.0% of total CLA in the diet. Total cis/cis isomers accounted for 3.8% total CLA in the diet but they were 6.6% of the total CLA in the egg yolk lipids. In contrast, total trans/trans isomers were 12.2% of the total CLA isomers in the egg yolk lipids, whereas they were only 4.2% of total CLA in the diet. The results showed that total trans/trans-CLA was preferentially incorporated into the egg yolk, whereas the incorporation of total cis/trans-CLA isomers was partially discriminated. Within each group, the incorporation of individual isomers into the egg yolk lipids was also selective. cis-9,trans-11/trans-9,cis-11 and cis-10,trans-12/trans-10,cis-12 were the two major isomers in the diet. Ag-HPLC analysis showed that the former was preferentially transferred into the egg yolk compared with the latter. It was observed that supplementation of CLA in the diet of laying hens decreased the concentration of oleic acid (18:1n-9), arachidonic acid (20:4n-6), and docosahexaenoic acid (22:6n-3) but increased that of linolenic acid (18:3n-3), stearic acid (18:0), and palmitic acid (16:0) in the egg yolk, suggesting that CLA may inhibit Delta6 and Delta9 desaturases.     

Ascorbic acid and ascorbic acid 6-palmitate induced oxidation in egg yolk dispersions

The oxidation in aqueous dispersions of egg yolk powder and the influence of addition of the proposed antioxidants ascorbic acid and ascorbic acid 6-palmitate indicate that both ascorbic acid and ascorbic acid 6-palmitate propagated the oxidation of egg yolk powder dispersions. Ascorbic acid 6-palmitate was found to be more prooxidative than ascorbic acid. Moreover, it was found that addition of ascorbic acid or ascorbic acid 6-palmitate gave rise to an increase in the amount of free iron Fe(II) in the egg yolk dispersions. It is proposed that ascorbic acid and ascorbic acid 6-palmitate react with the phosvitin-Fe(III) complex found in egg yolk and release Fe(II), which subsequently propagates lipid oxidation. It appears that less oxidation occurs in egg yolk dispersions exposed to high concentrations of peroxy radicals with added ascorbic acid than egg yolk dispersions with added ascorbic acid without exposure to peroxy radicals.     

Effect of short-term UVB exposure on vitamin D concentration of eggs and vitamin D status of laying hens

Vitamin D deficiency in humans is widespread, and only a few food items are important natural sources of vitamin D. This study investigated the effect of UVB exposure of laying hens on the vitamin D content in egg yolk. In a two-factorial design, hens fed a vitamin D-deficient (-D) or -adequate (+D) diet were nonexposed or exposed to UVB light over a period of 4 weeks. UVB exposure of the -D group caused nearly normal egg production rate and egg shell quality; exposure of the +D group did not further improve these parameters. UVB exposure tended to improve the concentration of plasma 25-hydroxycholecalciferol (25(OH)D(3)), but had no effect on 1,25-dihydroxycholecalciferol in plasma or on cholecalciferol and 25(OH)D(3) in egg yolk. The present study shows that a short-term exposure of laying hens to UVB light is not an appropriate way to improve the vitamin D content of egg yolk.     

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.     

Identification and quantification of astaxanthin esters in shrimp (Pandalus borealis) and in a microalga (Haematococcus pluvialis) by liquid chromatography-mass spectrometry using negative ion atmospheric pressure chemical ionization

Negative ion liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [negative ion LC-(APCI)MS] was used for the identification of astaxanthin esters in extracts of commercial shrimp (Pandalus borealis) and dried microalga (Haematococcus pluvialis) samples. A cleanup step using a normal phase solid phase extraction (SPE) cartridge was applied prior to analysis. Recovery experiments with astaxanthin oleate as model compound proved the applicability of this step (98.5 +/- 7.6%; n = 4). The assignment of astaxanthin esters in negative ion LC-(APCI)MS was based on the detection of the molecular ion (M*-) and the formation of characteristic fragment ions, resulting from the loss of one or two fatty acids. Quantification of individual astaxanthin esters was performed using an astaxanthin calibration curve, which was found to be linear over the required range (1-51 micromol/L; r2 = 0.9996). Detection limits, based on the intensity of M*-, a signal-to-noise ratio of 3:1, and an injection volume of 20 microL, were estimated to be 0.05 microg/mL (free astaxanthin), 0.28 microg/mL (astaxanthin-C16:0), and 0.78 microg/mL (astaxanthin-C16:0/C16:0), respectively. This LC-(APCI)MS method allows for the first time the characterization of native astaxanthin esters in P. borealis and H. pluvialis without using time-consuming isolation steps with subsequent gas chromatographic analyses of fatty acid methyl esters. The results suggest that the pattern of astaxanthin-bound polyunsaturated fatty acids of P. borealis does not reflect the respective fatty acid pattern found in triacylglycerides. Application of the presented LC-(APCI)MS technique in common astaxanthin ester analysis will forestall erroneous xanthophyll ester assignment in natural sources.     

基于计算机视觉的鸡蛋新鲜度无损检测

对鸡蛋的新鲜度、贮藏期进行无损检测,在鸡蛋生产、流通、加工领域具有重要意义。该文进行了鸡蛋新鲜度（哈夫值）常规指标在一定温度条件下随贮藏时间变化的试验,采用高分辨率的工业化数字摄像头,以冷光源背向照明方式（照度为10 000 Lx）获取数字图像,并提取了鸡蛋的图像特征蛋黄指数和气室指数。建立了鸡蛋新鲜度与蛋黄指数、贮藏时间与鸡蛋新鲜度、贮藏时间与蛋黄指数和气室指数的关系模型。得知鸡蛋新鲜度与蛋黄指数呈线性相关性,经检验实测值与预测值平均相对误差为6%;贮藏时间与鸡蛋新鲜度以及贮藏时间与鸡蛋蛋黄指数、鸡蛋气室指数均具有二次函数关系,经检验实测值与预测值绝对误差不超过2 d。结果表明基于计算机视觉技术、采用背向照明方式采集的鸡蛋的透射图像得到鸡蛋的蛋黄和气室的图像信息,可以预测鸡蛋的新鲜度和贮藏期。     

Carryover of maduramicin from feed containing cross-contamination levels into eggs of laying hens

Maduramicin is a coccidiostat authorized as feed additive in the European Union for chickens and turkeys for fattening but not for laying hens, considering the risk of residues in eggs. The unavoidable cross-contamination of non-target feed with coccidiostats is regulated by Commission Directive 2009/8/EC and resulting carry-over in food by Commission Regulation (EC) No. 124/2009. To verify the compliance of the maximum levels for maduramicin in feed (50 μg/kg) and eggs (2 μg/kg), the carry-over from feed into eggs was investigated. Diets containing 10, 30, and 50 μg of maduramicin/kg of feed were fed to laying hens. Feed, egg white, and yolk were analyzed by LC-MS/MS. Maduramicin residues were only detected in in egg yolk. Feeding the 10 μg/kg maduramicin diet resulted in maduramicin concentrations up to 2.5 μg/kg in whole eggs, already exceeding the maximum level. A carry-over rate of 8% maduramicin from feed into eggs was calculated.     

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) method extension to quantify simultaneously melamine and cyanuric acid in egg powder and soy protein in addition to milk products

As a consequence of the adulteration of infant formulas and milk powders with melamine (MEL) in China in 2008, much attention has been devoted to the analysis of MEL [and cyanuric acid (CA)] in dairy products. Several methods based on high-performance liquid chromatography (HPLC), liquid chromatography-tandem mass spectrometry (LC-MS/MS), nuclear magnetic resonance (NMR), or Raman spectroscopy have been described in the literature. However, no method is available for the simultaneous determination of MEL and CA in other raw materials, which are considered as high-risk materials for economically motivated adulteration. The present paper reports the results of an interlaboratory-based performance evaluation conducted with seven laboratories worldwide. The purpose was to demonstrate the ability of a cleanup-free LC-MS/MS method, originally developed for cow's milk and milk-powdered infant formula, to quantify MEL and CA in egg powder and soy protein. Limit of detection (LOD) and limit of quantification (LOQ) were 0.02 and 0.05 mg/kg for MEL in egg powder and soy protein, respectively. For CA, LOD and LOQ were 0.05 and 0.10 mg/kg in egg powder and 1.0 and 1.50 mg/kg in soy protein, respectively. Recoveries ranged within a 97-113% range for both MEL and CA in egg powder and soy protein. Reproducibility values (RSD(R)) from seven laboratories were within a 5.4-11.7% range for both analytes in the considered matrices. Horwitz ratio (HorRat) values between 0.4 and 0.7 indicate acceptable among-laboratory precision for the method described.     

Identification of benzalkonium chloride in commercial grapefruit seed extracts

Commercial grapefruit seed extracts (GSE) were extracted with chloroform. The solvent was evaporated, and the resulting solid was subsequently analyzed by high-performance liquid chromatography (HPLC), electrospray ionization mass spectrometry (ESI/MS), tandem mass spectrometry (ESI/MS/MS), and elemental analysis (by proton-induced X-ray emission analysis). Three major constituents were observed by HPLC and were identified as benzyldimethyldodecylammonium chloride, benzyldimethyltetradecylammonium chloride, and benzyldimethylhexadecylammonium chloride. This mixture of homologues is commonly known as benzalkonium chloride, a widely used synthetic antimicrobial ingredient used in cleaning and disinfection agents.     

基于透射图像的高压脉动腌制咸鸭蛋含盐量检测

含盐量是衡量咸鸭蛋品质的重要指标。为了利用机器视觉技术实现高压脉动腌制咸鸭蛋含盐量的无损检测。该研究采用工业相机和透射光源搭建咸鸭蛋的透射图像采集装置。采用图像整体特征和长轴截面光强度特征两种特征提取方法,利用多元线性回归、支持向量机回归两种算法,建立对蛋清、蛋黄及全蛋含盐量以及蛋黄指数的定量预测模型。结果表明,随着咸鸭蛋腌制时间的增加,其透光性显著提高。同时,透射图像蛋黄的所在视野区域会随着含盐量的增加而呈现规律性的变化。基于图像整体特征建立的蛋清、蛋黄、全蛋含盐量模型较优,在蛋黄指数预测下基于长轴截面光强度特征所建模型较优。其中,基于图像整体特征所建立的蛋黄含盐量支持向量机回归（support vector regression, SVR）模型最优,测试集相关系数（test set correlation coefficient, R_p）、测试集均方根误差(root mean square error of prediction, RMSE_p)、相对分析误差（residual predictive deviation, RPD）分别达到0.8460、0.3416、1.898;基于长轴截面光强度特征建立的蛋黄指数多元线性回归（multiple linear regression, MLR）模型最优,测试集相关系数R_p、均方根误差RMSE_p、相对分析误差RPD分别为0.8318、0.0743、1.916。该研究结果为咸鸭蛋含盐量的快速检测提供理论依据和技术支持。     

Enrichment of poultry products with omega3 fatty acids by dietary supplementation with the alga Nannochloropsis and mantur oil

Experiments were conducted to evaluate the efficiency of the microalga Nannochloropsis sp. (Nanno.), as a supplement to laying hens' diet, for the production of enriched eggs and meat with omega3 fatty acids (FA). Nanno. has a unique FA composition, namely, the occurrence of a high concentration of eicosapentaenoic acid (EPA; 20:5 omega3) and the absence of other omega3 FA. The effect of supplementing diets with Nanno. on omega3 FA levels in eggs, plasma, liver, and thigh muscle was compared to that of mantur oil, high in alpha-linolenic acid (LNA; 18:3 omega3). Nanno. is rich also in carotenoids, which may be useful for egg yolk pigmentation. The observed effect of Nanno. supplementation on yolk pigmentation was dose responsive, in both the rate of coloration and the color intensity. Addition of enzyme preparations (glucanase plus cellulase or glucanase plus pectinase) slightly elevated the yolk color score. The most prominent changes in the level of omega3 FA in egg yolk were evident when the diets were supplemented with 1% Nanno. or mantur lipid extracts. Levels of dietary algal meal (0.1-1.0%) had low and inconsistent effects on the level of yolk omega3 FA. Algal EPA is not accumulated in the liver or in the egg yolk; it is apparently converted and deposited as docosahexaenoic acid (DHA). LNA from mantur oil was partially converted to DHA, and both DHA and LNA were deposited in egg yolks and livers. It is suggested that the absence of DHA and EPA from thigh muscle is due to the small amount of dietary omega3 FA used in this work, compared to other studies, and to the possibility that in laying hens the egg yolk has a priority on dietary FA over that of muscles.     

Proteomics analysis of egg white proteins from different egg varieties

The market of specialty eggs, such as omega-3-enriched eggs, organic eggs, and free-range eggs, is continuously growing. The nutritional composition of egg yolk can be manipulated by feed diet; however, it is not known if there is any difference in the composition of egg white proteins among different egg varieties. The purpose of the study was to compare the egg white proteins among six different egg varieties using proteomics analysis. Egg white proteins were analyzed using two-dimensional gel electrophoresis (2-DE), and 89 protein spots were subjected to LC-MS/MS. A total of 23 proteins, belonging to Gallus gallus , were identified from 72 detected protein spots. A quiescence-specific protein precursor in egg white was identified for the first time in this study. Significant differences in the abundant levels of 19 proteins (from 65 protein spots) were observed among six egg varieties. Four proteins, ovalbumin-related protein Y, cystatin, avidin, and albumin precursor, were not different among these six egg varieties. These findings suggest that the abundance, but not the composition, of egg white proteins varied among the egg varieties.     

Impact of a treatment with phospholipase A2 on the physicochemical properties of hen egg yolk

Changes in physicochemical properties of egg yolk were investigated after a treatment with phospholipase A 2 (PLA 2), where phospholipids are converted in lyso-phospholipids. Protein solubility and protein denaturation before and after modification by PLA 2 was monitored as well as the functionality of egg yolk by means of interfacial tension. Enzymatic treatment showed a significant impact on the properties of egg yolk with regard to protein solubility and denaturation behavior. To gain a closer insight, egg yolk was separated in its water-soluble fraction called plasma and the insoluble granules. Both fractions were separately modified by PLA 2. The granule fraction shows a higher protein solubility, and the plasma proteins show very high heat stability after modification by PLA 2. Hypotheses regarding related changes in the low-density lipoprotein (LDL) particles are discussed. Results suggest that significant differences in the functional properties of untreated and PLA 2-modified egg yolk do not primarily result from the existence of lyso-phospholipids but from structural changes in egg yolk granules and LDL particles.