表面增强拉曼光谱法同时检测奶粉中三聚氰胺和二聚氰胺

本文建立了奶粉中同时检测三聚氰胺及二聚氰胺的表面增强拉曼光谱法.奶粉样品经15％三氯乙酸溶液提取,中性氧化铝吸附杂质后进行拉曼光谱检测.三聚氰胺线性范围为0.0050～0.075 mg/L,检出限为0.0015 mg/L,回收率在79.5％～124％之间,相对标准偏差小于8.8％(n=5);二聚氰胺线性范围为0.50～l0mg/L,检出限为0.15 mg/L,回收率在76.5％～112％之间,相对标准偏差小于9.4％(n=5).该方法相比常规表面增强拉曼光谱法,仅通过一种样品前处理手段即可对奶粉中的三聚氰胺或二聚氰胺进行定性检出及定量分析,相比色谱等检测方法具有样品前处理过程简单、耗时短等优点,在奶粉质量监控方面具有良好的应用前景.     

Application of FT-Raman Spectroscopy and Chemometric Analysis for Determination of Adulteration in Milk Powder

In this work, FT-Raman spectroscopy is explored as a rapid technique for the assessment of the milk powder quality. Based on information provided by Raman spectra of samples adulterated with starch and whey, a quantitative method is developed to identify the fraud, using Partial Least Squares regression (PLS). In regression models using PLS the results are satisfactory, and such models can be used to identify and quantify samples presenting whey and starch in milk powder at concentrations of 2.32% and 1.64% (w/w), respectively. In the whey determination, the obtained values in the PLS model of the new samples are compared with those obtained by the spectrophotometric method of acid ninhydrin. This result shows that there is no significant difference with the 95% level of confidence between the values provided by the PLS regression method and the acid ninhydrin. The present work shows Raman spectroscopy as an analytical tool which can be used in quality control of milk powder, even in fraud processes, and the calculated figures of merit such as sensitivity, accuracy, limit of detection and limit of quantification clearly demonstrate this potential use. Although the multivariate models developed are not strictly quantitative, especially for low concentrations, they can be used as screening methods for routine analysis, as showed by this work.     

固相萃取-表面增强拉曼光谱法测定奶粉中三聚氰胺

建立了固相萃取-表面增强拉曼光谱法(SPE-SERS)测定奶粉中三聚氰胺的分析方法。待测物经乙腈提取、活性碳吸附及氢氧化钠溶液洗脱后进行SERS检测。该方法的线性范围为0.005 0~1.6 mg/L,检出限为0.100 mg/kg,回收率为75.3%~125%,相对标准偏差(RSD,n=5)不大于9.3%。该法灵敏度高、杂质干扰小、结果准确可靠,能满足奶粉中三聚氰胺的快速检测,在奶粉质量监控方面具有良好的应用潜力。     

Determination of cyanuric acid using exact matching isotope dilution mass spectrometry

A high‐accuracy double isotope dilution mass spectrometric method using an exact matching approach with GC coupled to a mass spectrometer for the analysis of cyanuric acid in fortified milk powder was developed. Various parameters for sample clean‐up such as the type of SPE cartridge, GC column and type of derivatizing agent used were investigated. The method was found to be linear in the concentration range of 0.03 to 2 mg/kg of cyanuric acid in milk powder. LOD and LOQ were found to be 0.9 and 3 μg/kg, respectively. Recoveries in the range of 95.7 to 102.2% were obtained for the in‐house fortified milk powder samples, with RSD of measurements in the range of 0.2 to 3.0%. A metrological approach was undertaken to examine all possible biases that contributed to the combined measurement uncertainty of the method. This high‐accuracy method can serve as a reference method for techniques commonly applied in routine testing laboratories.     

Determination of Lactose in Milk by Raman Spectroscopy

A Raman spectroscopy-based approach was developed for the rapid determination of lactose in milk using crystal violet as an internal standard. The crystal violet peak at 1173 reciprocal centimeters did not interfere with the lactose peak at 1085 reciprocal centimeters; therefore, crystal violet can be used to quantify lactose in milk. This developed method allowed the determination of lactose from 0.028–0.1 mole per liter, with a limit of detection of 0.019 mole per liter. The results revealed that Raman spectroscopy is a simple and rapid method for the determination of lactose in milk.     

Effect of surface composition on the flowability of industrial spray-dried dairy powders

The surface composition of four industrial spray-dried dairy powders (skim milk powder, whole milk powder, cream powder and whey protein concentrate) was estimated by electron spectroscopy for chemical analysis (ESCA), and its influence on powder flowability was studied. It was found that skim milk powder flows well compared to the other powders because the surface is made of lactose and protein with a small amount of fat, whereas the high surface fat composition inhibits the flow of whole milk, cream and whey protein powders. However, the poor flowability of the powders with high surface fat coverage was drastically improved by removal of fat present on the surface through a brief wash with petroleum ether. The results obtained indicate that, although there are several parameters including particle size, which influence the flowability of powders, the flowability of powders is strongly influenced by the surface composition of powders, particularly for fat-containing powders.     

Determination of fat, protein, casein, total solids, and somatic cell count in goat's milk by near-infrared reflectance spectroscopy

Analysis using near-infrared reflectance spectroscopy (NIRS) was investigated as a means of predicting quality parameters of goat's milk. Calibration equations were developed with samples from individual goats of Malague?a and Murciano-Granadina dairy breeds at different stages of lactation to obtain a wide range of variation in milk composition. Prediction equations for milk fat, protein, casein, total solids, and somatic cell count (SCC) were developed with 2 sample presentations, liquid drawer (homogenized milk) and aluminium cup (unhomogenized milk), to measure absorbance values, and accuracy of measurements was compared. The multiple correlation coefficient (R) values for homogenized milk were 0.98, 0.96, 0.91, 0.94, and 0.79 and for unhomogenized milk were 0.98, 0.95, 0.92, 0.95, and 0.74 for fat, protein, casein, total solid, and SCC, respectively. To validate the calibrations, an independent set of samples was used. The best simple correlation coefficient (r) values were 0.97, 0.95, 0.91, 0.93, and 0.72 for homogenized milk and 0.97, 0.95, 0.92, 0.95, and 0.79 for unhomogenized milk. The study showed that NIRS is a useful technique for the prediction of fat, protein, casein, total solids, and SCC in unhomogenized goat's milk.     

Determination of total selenium and selenium distribution in the milk phases in commercial cow’s milk by HG-AAS

A procedure has been developed for determining the selenium in cows milk using hydride generation–atomic absorption spectrometry (HG-AAS) following microwave-assisted acid digestion. The selenium distributions in milk whey, fat and micellar casein phases were studied after separating the different phases by ultracentrifugation and determining the selenium in all of them. The detection limits obtained by HG-AAS for the whole milk, milk whey and micellar casein were 0.074, 0.065 and 0.075 g l^–1, respectively. The accuracy for the whole milk was checked by using a Certified Reference Material CRM 8435 whole milk powder from NIST, and the analytical recoveries for the milk whey and casein micelles were 100.9 and 96.9%, respectively. A mass balance study of the determination of selenium in the different milk phases was carried out, obtaining values of 95.5–100.8%. The total content of selenium was determined in 37 milk samples from 15 different manufacturers, 19 whole milk samples and 18 skimmed milk samples. The selenium levels found were within the 8.5–21 g l^–1 range. The selenium distributions in the different milk phases were studied in 14 whole milk samples, and the highest selenium levels were found in milk whey (47.2–73.6%), while the lowest level was found for the fat phase (4.8–16.2%). A strong correlation was found between the selenium levels in whole milk and the selenium levels in the milk components.     

Determination of vegetal proteins in milk powder by sodium dodecyl sulfate-capillary gel electrophoresis: interlaboratory study

An interlaboratory study, with the participation of 8 laboratories, was conducted to evaluate a sodium dodecyl sulfate-capillary gel electrophoresis method for determination of adulteration of milk powder with soy and pea proteins. Calibration standards (0-8%, w/w, soy and pea protein in total protein) and adulterated skim milk powders (0-5%, w/w, soy and pea proteins in total protein) were produced. Vegetal proteins were determined after removal of milk proteins by pretreatment of the samples with tetraborate-EDTA buffer, pH 8.3. Repeatability standard deviations ranged from 9 to 15% and reproducibility standard deviations ranged from 25 to 30% in the samples containing 5% vegetal protein in total protein.     

毛细管电泳检测奶粉中添加的大豆分离蛋白

采用毛细管电泳方法对乳蛋白和大豆分离蛋白进行检测。选择聚丙烯酰胺涂层毛细管,在紫外检测波长214 nm、分离电压25 kV条件下测出五种乳蛋白在不同线性范围内的线性相关系数均大于0.998,各乳蛋白的迁移时间和峰面积的相对标准偏差(RSD)分别小于0.34%和4.50%,奶粉样品加标回收率范围为88.07%~102%;通过对比大豆7S和11S组分、大豆分离蛋白及乳蛋白的电泳图谱,定性确定出大豆分离蛋白8个特征峰,各特征峰的迁移时间和峰面积的RSD分别小于0.36%和4.87%,方法重现性较好。建立了奶粉中添加大豆分离蛋白的半定量检测方法。     

Fast determination of milk fat content using Raman spectroscopy

In our work, we have demonstrated the capability of VIS Raman spectroscopy in combination with partial least square regression (PLS) as a rapid technique for direct milk fat determination. Raman spectra of milk samples revealed contributions from proteins, but mainly from their fat content with different spectral characteristics. Three different methods of sample preparations were applied: (i) liquid milk contained in an open dish, (ii) dried milk droplets on glass plates covered with Al foil, and (iii) liquid milk contained in quartz cuvettes. Methods (i) and (ii) showed a good PLS model for milk fat prediction with low root mean square errors and high correlation coefficients. The main advantage of milk sample contained in the dish lies in its simplicity as well as the fact that the open container maximizes the signal of interest avoiding background contributions. Our results show that Raman spectroscopy is suited for in-line monitoring purposes.     

在线透析-流动注射光度法测定奶及奶制品中尿素含量

建立了在线透析-流动注射光度法测定奶及奶制品中尿素的方法.加人三氯乙酸使样品中蛋白质沉淀,离心过滤去除沉淀的蛋白质,在线透析去除剩余的脂肪及大分子,避免干扰光度分析.在Fe3+存在时,尿素与二乙酰一肟在酸性溶液中水解生成的二乙酰反应生成红色化合物,在534 nm处测量吸光度,加入硫代氨基脲提高显色强度.实验对试剂的浓度...     

Determination of polybrominated diphenyl ethers in milk samples. Development of green extraction coupled techniques for sample preparation

Ultrasound‐assisted extraction (UAE), cloud point extraction (CPE), and ultrasound back‐extraction (UABE) techniques have been coupled for lixiviation, preconcentration, and cleanup of polybrominated diphenyl ethers (PBDEs) from milk samples for determination by gas chromatography‐electron capture detection (GC‐ECD). Physicochemical parameters that affect the efficiency of the extraction system were investigated using a design of experiments based on multivariate statistical tools, and considering the sample matrix along the development. The coupling of the leaching step, UAE, enhanced ca. 3.5 times the extraction efficiency of the former sample preparation methodology (CPE‐UABE) leading to cleaner sample extracts suitable for GC analysis. Under optimum conditions, the proposed methodology exhibits successful performance in terms of linearity and precision, with recoveries in the range of 68–70% and LODs within the range 0.05–0.5 ng/g dry weight (d.w.). The proposed sample preparation methodology coupled three green analytical techniques. It expands the application frontiers of CPE for the analysis of biological samples by GC. The optimized methodology was used for determination of PBDEs in powder milk samples, from both commercial and human sources.     

On-Site Measurement of Fat and Protein Contents in Milk Using Mobile NMR Technology

Robust and easy-to-use NMR sensor technology is proposed for accurate, on-site determination of fat and protein contents in milk. The two parameters are determined using fast consecutive ¹H and ³⁵Cl low-field NMR experiments on milk samples upon the 1:1 addition of a low-cost contrast solution. Reliable and accurate measurements are obtained without tedious calibrations and the need for extensive database information and may readily be conducted by non-experts in production site environments. This enables on-site application at farms or dairies, or use in laboratories harvesting significant reductions in costs and time per analysis as compared to wet-chemistry analysis. The performance is demonstrated for calibration samples, various supermarket milk products, and raw milk samples, of which some were analyzed directly in the milking room. To illustrate the wide application range, the supermarket milk products included both conventionally/organically produced, lactose-free milk, cow’s, sheep’s and goat’s milk, homogenized and unhomogenized milk, and a broad nutrient range (0.1–9% fat, 1–6% protein). Excellent agreement between NMR measurements and reference values, without corrections or changes in calibration for various products and during extensive periods of experiment conduction (4 months) demonstrates the robustness of the procedure and instrumentation. For the raw milk samples, correlations between NMR and IR, NMR and wet-chemistry, as well as IR and wet-chemistry results, show that NMR, in terms of accuracy, compares favorably with the other methods.     

Detection of the presence of soya protein in milk powder by sodium dodecyl sulfate capillary electrophoresis

Two different commercial kits for sodium dodecyl sulfate capillary electrophoresis (SDS-CE) were evaluated for the detection of the presence of soya protein in milk powder. The results obtained showed that SDS-CE allowed the separation of the basic subunits of glycinin and the alpha and alpha' subunits of beta-conglycinin from the main milk protein peaks. However, a detection limit lower than 10% (w/w) of soya protein in total protein could not be achieved. The use of a tetraborate-EDTA sample treatment minimized interferences from milk proteins, allowing the detection of at least 1% (w/w) of soya protein in total protein. The addition of soya protein hydrolysates could not be determined using SDS-CE.     

Use of NIRS technology with a remote reflectance fibre-optic probe for predicting major components in cheese

In the present work the potential of near infra-red spectroscopy technology (NIRS) together with the use of a remote reflectance fibre-optic probe for the analysis of fat, moisture, protein and chlorides contents of commercial cheeses elaborated with mixtures of cow's, ewe's and goat's milk and with different curing times was examined. The probe was applied directly, with no previous sample treatment. The regression method employed was modified partial least squares (MPLS). The equations developed for the cheese samples afforded fat, moisture, protein, and chloride contents in the range 13-52%, 10-62%, 20-30%, and 0.7-2.9%, respectively. The multiple correlation coefficients (RSQ) and prediction corrected standard errors (SEP (C)) obtained were respectively 0.97 and 0.995% for fat; 0.96% and 1.640% for moisture; 0.78% and 0.760% for protein, and 0.89% and 0.112% for chlorides.     

Raman spectroscopy based analysis of milk using random forest classification

The development of a classification system based on the Raman spectra of milk samples is proposed in present study. Such development could be useful for nutritionists in suggesting healthy food to infants for their proper growth. Previously, molecular structures in milk samples have been exploited by Raman spectroscopy. In the current study, Raman spectral data of milk samples of different species is utilized for multi-class classification using a dimensionality reduction technique in combination with random forest (RF) classifier. Quantitative and experimental analysis is based on locally collected milk samples of different species including cow, buffalo, goat and human. This classification is based on the variations (different concentrations of the components present in milk such as proteins, milk fats, lactose etc.) in the intensities of Raman peaks of milk samples. Principal component analysis (PCA) is used as a dimensionality reduction technique in combination with RF to highlight the variations which can differentiate the Raman spectra of milk samples from different species. The proposed technique has demonstrated sufficient potential to be used for differentiation between milk samples of different species as the average accuracy of about 93.7%, precision of about 94%, specificity of about 97% and sensitivity of about 93% has been achieved.     

电感耦合等离子体发射光谱法同时测定奶粉中钙铜铁钾镁锰钠锌和磷

通过优化试验确定仪器的最佳工作参数,微波加热硝酸消解样品,采用电感耦合等离子体发射光谱法同时测定奶粉中钙、铜、铁、钾、镁、锰、钠、锌和磷9种元素的含量。分析了4种奶粉样品,所得结果与国标方法的结果一致。方法回收率在96.5%-111%之间。     

液相色谱-质谱联用法测定牛奶中14种青霉素及相应青霉噻唑酸残留量

利用超高效液相色谱-串联质谱法(UPLC-MS/MS)同时测定牛奶中的7种青霉素类抗生素以及7种相应的青霉噻唑酸。样品经乙腈沉淀蛋白,上清液N2吹干后,用水溶解,加入正己烷萃取除去脂肪;提取液经ACQUITY UPLCBEH C18柱分离,乙腈-乙酸铵+甲酸水溶液洗脱。14种物质峰分离良好,定量限范围在5～20μg/kg。在10～50ng/mL质量浓度范围内线性良好,相关系数均大于等于0.999,牛奶中的加标回收率在90%～98%。     

奶粉中唾液酸的测定方法

利用酸水解把奶粉中的唾液酸释放出来,通过聚丙烯酰胺凝胶除去奶粉中大量乳糖的干扰。利用Cu^2＋和Cr^3＋协同催化唾液酸与间苯二酚的显色,用乙酸丁酯-正丁醇萃取有色物质,在610nm处测定牛奶中唾液酸含量。方法的线性范围0～55．90mg／L,回收率为94％～96％。