羊肉纯度电子舌快速检测方法

为实现掺假羊肉的快速、客观评价,利用电子舌对混入不同比例鸡肉的掺假羊肉糜进行检测及定性和定量分析。3种浸提溶液分别浸提,样品量均对电子舌传感器的响应影响极显著;以数据点重复性和聚类效果为依据,采用主成分分析方法确定了电子舌检测羊肉糜样品的较佳条件为0.1 mol/L KCl溶液浸提15 g肉糜样品。在此较佳条件下,对混入不同比例鸡肉的掺假羊肉进行检测,结果表明：采用主成分分析和典则判别分析,前2个主成分累积贡献率均超过80%,电子舌均能很好地区分混入不同比例鸡肉的羊肉糜样品;采用多元线性回归分析和偏最小二乘回归分析建立的定量预测模型能有效预测混入的鸡肉比例（R2>0.99,RMSE<3%）。试验表明：电子舌在羊肉掺入鸡肉的鉴别中具有可行性,研究结果可为羊肉掺假鉴别提供参考。     

Detection of pork and lard as adulterants in processed meat: liquid chromatographic analysis of derivatized triglycerides

A new method is described for detection of pork and lard as adulterants in processed beef and mutton mixtures. The unsaturated triglycerides in the fat are ozonized and then derivatized. The mixture of derivatized and saturated triglycerides is analyzed by liquid chromatography using a reverse-phase column and a UV detector. Pork fat has larger amounts of triglyceride containing saturated fatty acid at the C-2 position than does the fat of other meat. The ratio of triglyceride containing saturated fatty acid vs triglyceride containing unsaturated fatty acid at the same (C-2) position (SSU/SUS) in a sample is compared with those of pure meats. The presence of pork in the sample causes the ratio to increase compared with ratios for pure beef or mutton. The increase in the SSU/SUS ratio is significant for the addition of 1% pork in beef. In the case of mutton, the addition of 3% pork causes a noticeable change. The method is reliable and is also applicable to samples containing only fat. Processing (heating or cooking) does not affect the ratios.     

基于生物散斑图像和惯性矩谱分析的牛肉掺腐检测

牛肉掺假严重危害消费者的健康与经济利益,因此对牛肉掺假进行无损检测具有重要意义。该文基于生物散斑技术对牛肉掺假进行定量检测。试验将新鲜牛肉和非新鲜牛肉按不同比例(0、1%、3%、5%~60%(5%梯度)和100%)混合制备掺假样本,并采集样本的生物散斑图像。针对单列惯性矩(inertia moment,IM)表征样本生物活性存在稳定性差的问题,首次提出惯性矩谱(IM谱)分析的方法并用于建立基于支持向量回归机(support vector regression machine,SVR)的牛肉掺假检测模型。结果表明基于IM谱建立的SVR模型能较为准确预测牛肉中掺假物含量,校正集和测试集的决定系数分别为0.85和0.81,均方根误差分别为0.12和0.11。该研究证明了利用生物散斑技术和惯性矩谱分析方法对新鲜牛肉中掺杂腐败牛肉进行定量检测是可行的。     

Authentication of animal fats using direct analysis in real time (DART) ionization-mass spectrometry and chemometric tools

A combination of direct analysis in real time (DART) ionization coupled to time-of-flight mass spectrometry (TOFMS) and chemometrics was used for animal fat (lard and beef tallow) authentication. This novel instrumentation was employed for rapid profiling of triacylglycerols (TAGs) and polar compounds present in fat samples and their mixtures. Additionally, fat isolated from pork, beef, and pork/beef admixtures was analyzed. Mass spectral records were processed by principal component analysis (PCA) and stepwise linear discriminant analysis (LDA). DART-TOFMS profiles of TAGs were found to be more suitable for the purpose of discrimination among the examined fat types as compared to profiles of polar compounds. The LDA model developed using TAG data enabled not only reliable classification of samples representing neat fats but also detection of admixed lard and tallow at adulteration levels of 5 and 10% (w/w), respectively. The presented approach was also successfully applied to minced meat prepared from pork and beef with comparable fat content. Using the DART-TOFMS TAG profiles of fat isolated from meat mixtures, detection of 10% pork added to beef and vice versa was possible.     

高光谱图像结合特征变量筛选定量检测羊肉中狐狸肉掺假

为了探讨快速无损检测羊肉糜中狐狸肉掺假含量的可行性,该研究利用高光谱技术结合特征变量筛选方法开展了其定量检测研究。利用遗传算法、竞争性自适应重加权算法和二维相关光谱分析（Two-Dimensional Correlation Spectroscopy,2D-COS）3种方法分别对代表性样品全部846个波长进行特征波长筛选,得到207、34和14个特征波长;基于全部波长和特征波长建立羊肉糜中狐狸肉掺假含量的偏最小二乘回归（Partial Least Squares Regression,PLSR）和支持向量回归（Support Vector Regression,SVR）模型并进行比较。研究结果表明,基于全部波长和特征波长建立的SVR模型性能均优于PLSR模型。其中,利用2D-COS方法提取的14个特征波长建立的SVR模型（即2D-COS-SVR模型）性能最优,其预测集决定系数和均方根误差分别为0.928和3.00%,相对分析误差为4.85,表明高光谱结合2D-COS-SVR模型可以有效实现羊肉糜中狐狸肉掺假的定量检测。该研究结果为开发低成本肉类掺假检测系统提供技术支持和参考依据。     

Use of polymorphisms in the γ-gliadin gene of spelt and wheat as a tool for authenticity control

Partial sequencing of the γ-gliadin gene of 62 spelt and 14 soft wheat cultivars was performed. Fifty-six of the 62 spelt cultivars and 13 of the 14 soft wheat cultivars were shown to exhibit the typical spelt or soft wheat γ-gliadin sequence, respectively. Exceptions were ascribed to crossbreeding of soft wheat and spelt. Using the typical soft wheat γ-gliadin sequence, two alternative DNA-based analytical methods were developed for the detection and quantification of spelt flour "adulteration" with soft wheat. A simple and fast detection of soft wheat in spelt flours could be achieved by restriction fragment length (RFLP) analysis. In combination with lab-on-a-chip capillary gel electrophoresis (LOC-CE) the soft wheat proportion could be estimated. Heteroduplex formation served as additional confirmation for the presence of spelt besides soft wheat. Hence, RFLP-LOC-CE constitutes a perfect analysis tool for the quality control of cereal seeds and pure cultivars. A precise quantification of soft wheat "adulterations" in spelt flour down to 1% could be achieved by the developed real-time PCR method. The calibration parameters of the real-time PCR assay fulfilled the minimum performance requirements of the European Network of GMO (genetically modified organisms) Laboratories (ENGL).     

电子鼻在牦牛肉和牛肉猪肉识别中的应用

为了探索电子鼻对肉类掺假识别的可行性,利用电子鼻对牦牛肉、牛肉和猪肉样品进行了分析。通过对所获得的数据进行主成分分析（principal component analysis,PCA）、判别因子分析（discriminant factor analysis,DFA）和偏最小二乘回归分析（partial least-squares analysis,PLS）。结果表明：几种肉类在电子鼻传感器上有不同的特征性响应图谱,电子鼻能够有效识别猪、牛肉;同时电子鼻能够识别不同部位的牦牛肉和普通牛肉;但不能识别不同部位的猪肉。在牛肉馅中掺入不同比例的猪肉馅时,电子鼻也能进行识别。采用偏最小二乘回归分析对数据进行处理,电子鼻响应信号和猪肉馅掺入比例之间有很好的相关性（决定系数R2为0.9762）,PLS模型预测误差在1.27%～7.00%之间。试验证明电子鼻可用于肉类的识别。     

中国草食牲畜发展战略

未来30年,中国人口15亿时,人口增加需要增加口粮0.4亿t;按现在肉食品生产水平和生产方式发展,届时中国至少需要增加猪肉禽肉蛋生产0.12亿t,需要增加饲料粮0.31亿t,总体增加0.71亿t,加之奶生产增加所需增加的饲料粮,粮食总生产需要达到5.71亿t以上,这是很难实现的目标。改变肉食品生产方式,即减少猪肉生产增加牛羊肉生产,具有节约300亿kg粮食的潜力;提高草食牲畜饲料转化率至5%~6%,并将多生产的牛羊肉替换猪肉生产,具有节约粮食200亿kg的潜力,两项合计可以节约粮食500亿kg。本文分析了发展节粮型草地畜牧业,节约粮食消耗,并保证肉品供应的途径,提出奶类生产通过发展1亿只奶山羊替换1670万头奶牛,以解决农村人口喝奶问题,实现全国奶生产消费与现今世界水平持平的目标。为实现草食牲畜生产增加2倍的目标,需要充分挖掘作物秸秆资源及南方冬闲田、疏林地、草原区的隐域生境等土地资源,同时提高草地和上述资源的优化利用效率;大力发展豆科牧草资源,解决中国粗饲料蛋白含量不足以及调整牲畜分布格局也是关键问题。     

Quantitative PCR detection of pork in raw and heated ground beef and pâté

Quantitative estimates are important to establish whether pork adulteration in ground beef and paté is accidental or intentional. A PCR procedure has been developed and evaluated to quantify pork in heated and nonheated meat and patés by densitometry using a specific and sensitive repetitive DNA element. Thirty, twenty-five, and twenty PCR cycles were carried out to find the best standard curve and correlation between pork content and band intensity. Twenty cycles showed the best results, quantifying degree contamination up to 1% pork in beef (heated and nonheated) and pork in duck paté with a minimum error. Finally, fraud was found in commercial patés.     

Compared use of HPLC and FZCE for cluster analysis of Triticum spp and for the identification of T. durum adulteration

Wheat quality criteria continually evolve in response to market pressure and consumer preference. Characterization of cereal cultivars for quality and agronomic properties, have widely shown the importance of the protein content to ensure good quality products. The aim of this work is a comparison of reversed-phase high performance liquid chromatography (RP-HPLC) and free zone capillary electrophoresis (FZCE) in the identification of Italian wheat cultivars and detection of durum wheat flour adulteration. Mainly alcohol soluble (gliadins) and water soluble (albumins) proteins were extracted from 14 common wheat cultivars and from 9 durum wheat cultivars. In RP-HPLC chromatograms, wheat albumins and gliadins eluted between 3 and 9 min and between 10 and 42 min, respectively. Even if the chosen chromatographic conditions (reversed phase) did not permit a complete resolution of hydrophilic proteins such as albumins, a good reproducibility was observed for both albumins and gliadins. In FZCE electropherograms, wheat albumins and gliadins migrated between 8 and 14 min and 16-25 min, respectively. A good reproducibility was found for wheat albumins, while the relatively poor reproducibility of gliadin fractions was a consequence of the selected separation conditions aimed to separate in the same run either hydrophilic (albumins) and alcohol-soluble (gliadins) proteins. The principal component analysis (PCA) of HPLC and FZCE data evidenced that both techniques allowed the univocal identification of the great proportion of investigated wheat cultivars. Three peaks were exclusively detected in RP-HPLC chromatograms of common wheat cultivars, while three unique peaks were found in FZCE electropherograms of common wheat cultivars. These peaks were investigated as a basis for detecting and estimating the adulteration of durum wheat flour with flour from common wheat. The direct relationship between the area of the peaks and adulteration level enabled standard curves to be constructed. The standard curves showed that adulteration may be quantified by either RP-HPLC or FZCE.     

Detection of soft wheat in semolina and durum wheat bread by analysis of DNA microsatellites

The aim of this work was to evaluate the analysis of DNA microsatellites for the detection of soft wheat (Triticum aestivum L.) in semolina and durum wheat bread (prepared from Triticum turgidum L. var. durum). The results enabled selection of an efficient D-genome-specific repetitive DNA sequence to detect common wheat in semolina and breads by qualitative PCR with a threshold of 3 and 5%, respectively, lowered to 2.5% by real-time PCR. This is of major importance for checking during production of some typical products recently awarded the European Protected Designation of Origin (PDO) mark such as Altamura bread, which should not contain soft wheat flour. The feasibility of quantification of common wheat adulteration in semolina using real-time PCR was also demonstrated.     

Textural Comparisons of Gluten‐Free and Wheat‐Based Doughs,Batters, and Breads

Studies were conducted with two newly developed gluten‐free bread recipes. One was based on corn starch (relative amount 54), brown rice (25), soya (12.5), and buckwheat flour (8.5), while the other contained brown rice flour (50), skim milk powder (37.5), whole egg (30), potato (25), and corn starch (12.5), and soya flour (12.5). The hydrocolloids used were xanthan gum (1.25) and xanthan (0.9) plus konjac gum (1.5), respectively. Wheat bread and gluten‐free bread made from commercial flour mix were included for comparison. Baking tests showed that wheat and the bread made from the commercial flour mix yielded significantly higher loaf volumes (P < 0.01). All the gluten‐free breads were brittle after two days of storage, detectable by the occurrence of fracture, and the decrease in springiness (P < 0.01), cohesiveness (P < 0.01), and resilience (P < 0.01) derived from texture profile analysis. However, these changes were generally less pronounced for the dairy‐based gluten‐free bread, indicating a better keeping quality. Confocal laser‐scanning microscopy showed that the dairy‐based gluten‐free bread crumb contained network‐like structures resembling the gluten network in wheat bread crumb. It was concluded that the formation of a continuous protein phase is critical for an improved keeping quality of gluten‐free bread.     

Influence of Milk,Corn Starch,and Baking Conditions on the Starch Digestibility,Gelatinization, and Fracture Stress of Biscuits

Dough for nontraditional semisweet biscuits—prepared with wheat flour or replacing part of the wheat flour with corn starch, with or without skim milk—was baked at two oven temperatures, 120 or 170°C, until reaching moisture content and water activity lower than 6% and 0.5, respectively. Assays of fracture stress, differential scanning calorimetry, X‐ray diffraction, and starch digestibility were performed. Results showed that biscuits containing milk had the highest fracture stress, and biscuits baked at low temperature were harder than biscuits baked at high temperature. The degree of starch gelatinization during baking was higher when dough was baked at 170°C, compared with dough baked at 120°C. The decrease in gelatinization coincides with the decrease in the height and surface of peaks at 15 and 23° in the X‐ray diffraction patterns. Milk and corn starch did not affect the starch digestibility of biscuits, but biscuits baked at 170°C presented lower fracture stress and higher starch digestibility than biscuits baked at 120°C.     

Sample preparation and liquid chromatographic determination of vitamin D in food products

Vitamin D in different fortified foods is determined by using liquid chromatography (LC). Sample preparation is described for fortified skim milk, infant formulas, chocolate drink powder, and diet food. The procedure involves 2 main steps: saponification of the sample followed by extraction, and quantitation by LC analysis. Depending on the sample matrix, additional steps are necessary, i.e., enzymatic digestion for hydrolyzing the starch in the sample and cartridge purification before LC injection. An isocratic system consisting of 0.5% water in methanol (v/v) on two 5 microns ODS Hypersil, 12 X 0.4 cm id columns is used. Recovery of vitamin D added to unfortified skim milk is 98%. The results of vitamin D determination in homogenized skim milk, fortified milk powder, fortified milk powder with soybean, chocolate drink powder, and sports diet food are given.     

基于近红外光谱技术的蜂蜜掺假识别

为了实现蜂蜜掺假的快速识别,应用近红外光谱结合模式识别方法对蜂蜜掺假现象进行了识别分析。该研究收集了中国不同品种、不同地域的典型天然蜂蜜样品,根据目前市场上常见的蜂蜜掺假手段,掺假物质及相对含量情况配制了掺假蜂蜜样品,利用傅立叶近红外光谱仪采集其透反射近红外光谱,分别采用偏最小二乘判别分析（PLS-DA）,独立软模式法（SIMCA）,误差反向传播神经网络（BP-ANN）和最小二乘支持向量机（LS-SVM）等模式识别方法,进行蜂蜜掺假识别研究。研究结果表明：利用这4种方法在蜂蜜中掺入果葡糖浆和果葡糖水的情况下均能很好地识别出掺假蜂蜜样品,其中对于掺入果葡糖浆的掺假情况,校正集的正确判别率均达到95%以上,验证集的正确判别率均达到87%以上,对于掺入果葡糖水的掺假蜂蜜校正集的正确判别率均达到93%以上,验证集的正确判别率均达到84%以上。通过比较4种不同的识别算法,发现采用LS-SVM时,对两种掺假情况下校正集和验证集的正确判别率均达到了100%,表明基于近红外光谱的蜂蜜掺假快速准确识别是可行的。     

Volumetric method for determing traces of organic chlorine in lipids.

The official AOAC method for determining chlorine in cake flour lipids has been modified to determine chlorine in lipids of animal carcasses cooled with chlorinated water. The sensitivity of the method has been improved 1000-fold to attain a detection limit of 2 ppm Cl in lipids. Recovery of the fine silver chloride precipitate was improved through centrifugation rather than filtration. The official method also did not take into consideration that when the silver chloride precipitate is exposed to light, it is converted to free chlorine and silver. Recovery of organic chlorine ranged from 84.6% at 4.1 ppm to 100.1% at 100 ppm. A number of samples of commercially chlorinated beef, pork, and chicken and laboratory-chlorinated chicken were analyzed by this method. In all cases where the level of chlorination was sufficient to result in a chlorine level in lipids in excess of 2 ppm, the chlorine content of the carcass lipids was measurable.     

牛肉质构特性的近红外光谱无损检测

为了建立基于近红外光谱技术的牛肉质构特性快速检测方法,该试验采集了202个新鲜牛肉样品在800～2500 nm波长范围内的漫反射光谱,测定了牛肉的硬度、弹性、咀嚼性和黏附性,经小波消噪后,分别采用平滑、一阶微分、二阶微分等6种方法预处理,建立了牛肉质构特性的偏最小二乘回归模型,并用最优模型进行预测。结果表明：经小波消噪后采用二阶微分预处理方法建立的牛肉硬度、弹性、咀嚼性的检测模型效果最好,其校正集相关系数 r 均在0.9以上,校正集均方根误差（root means square error of calibration,RMSEC）分别为6.247 N、0.760 mm、14.954 mJ,预测集相关系数均在0.664以上,预测集均方根误差（root means square error of prediction,RMSEP）分别为8.887 N、0.951 mm、22.117 mJ,相对预测误差（ratio of prediction to deviation,RPD）值分别为2.43、1.88、2.32,预测精度较高,能够有效地预测牛肉的硬度、咀嚼性,可以检测精度要求不高的牛肉弹性;试验所建立的牛肉黏附性检测模型的预测性能不是很理想,虽然其校正集和预测集相关系数较高（分别为0.720、0.694）,RMSEC 和 RESEP 均较小（分别为0.302、0.243 N·mm）,但其 RPD 值小于1.5,模型预测精度较差,不可以用于预测未知样品的黏附性,此方法还需进一步研究。研究结果为牛肉质构特性的快速无损评价提供了理论依据。     

二维相关光谱结合偏最小二乘法测定牛奶中的掺杂尿素

为了检验牛奶中是否掺杂尿素并将其量化测定,配置含有尿素质量浓度范围为1～20g/L之间40个牛奶样品,以掺杂物尿素浓度为外扰,分别研究了掺杂尿素牛奶的二维相关(近红外-近红外,中红外-中红外,近红外-中红外)光谱特性,在此基础上,分别选择随浓度变化大的4200～4800cm-1和1400～1704cm-1为建模区间,采用偏最小二乘方法建立定量分析模型。研究结果表明:4200～4800cm-1建模分析效果优于1400～1704cm-1建模结果,其交叉验证均方根误差为0.266g/L,对未知样品集预测相关系数达到0.999,预测均方根误差为0.219g/L,这表明所建模型具有较好的预测效果。该方法无需样品处理,成本低,为快速判别牛奶是否掺杂提供了一种新的可能的方法。     

Application of Response Surface Methodology in the Development of Gluten‐Free Bread

The formulation of gluten‐free (GF) bread of high quality presents a formidable challenge as it is the gluten fraction of flour that is responsible for an extensible dough with good gas‐holding properties and baked bread with good crumb structure. As the use of wheat starch in GF formulations remains a controversial issue, naturally GF ingredients were utilized in this study. Response surface methodology was used to optimize a GF bread formulation primarily based on rice flour, potato starch, and skim milk powder. Hydroxypropylmethylcellulose (HPMC) and water were the predictor variables. Analyses of the treatments from the design were made 24 hr after baking. Specific volume and loaf height increased as water addition increased (P < 0.01). Crumb firmness decreased as water levels increased (P < 0.01). Significant interactions (P < 0.01) between HPMC and water were found for the number of cells/cm². The number of large cells (>4 mm²) decreased with increasing levels of HPMC and water. Optimal ingredient levels were determined from the data obtained. The optimized formulation contained 2.2% HPMC and 79% water flour/starch base (fsb) and measured responses compared favorably to predicted values. Shelf‐life analysis of the optimized formulation over seven days revealed that, as crumb firmness increased, crust firmness and crumb moisture decreased.     

Direct optical biosensor analysis of folate-binding protein in milk

An automated, rapid, sensitive, and label-free biosensor-based assay for folate-binding protein (FBP) in bovine milk utilizing surface plasmon resonance optical detection is described. The active concentration of FBP is estimated from its specific interaction with a pteroyl-l-glutamic (folic) acid derivative immobilized on the sensor surface in a direct binding assay format. Milk, colostrum, and milk powders are prepared for analysis by dilution into buffer. Analysis conditions, including ligand immobilization, flow rate, contact time, and regeneration, have been defined, and nonspecific binding considerations were evaluated. Performance parameters include a working range for FBP in buffer of 0-200 ng/mL, a method detection limit of 0.13 microg/mL in fluid milk, overall instrument response RSD(R) of 0.64%, a mean interassay RSD(R) of 7.3% for skim milk powder, and surface stability over ca. 200 samples. The technique was applied to the measurement of active FBP content of consumer milks, the heat classification of skim milk powders manufactured under a wide range of thermal processing protocols, and change during early bovine lactation.