Detection of banned meat and bone meal in feedstuffs by near-infrared microscopic analysis of the dense sediment fraction

In this paper we present an alternative method for detection of meat and bone meal (MBM) in feedstuffs by near-infrared microscopic (NIRM) analysis of the particles in the sediment fraction (dense fraction (d >1.62) from dichloroethylene) of compound feeds. To apply this method the particles of the sediment fraction are spread on a sample holder and presented to the NIR microscope. By using the pointer of the microscope the infrared beam is focussed on each particle and the NIR spectrum (1112–2500 nm) is collected. This method can be used to detect the presence of MBM at concentrations as low as 0.05% mass fraction. When results from the NIRM method were compared with the classical microscopic method, a coefficient of determination (R²) of 0.87 was obtained. The results of this study demonstrated that this method could be proposed as a complementary tool for the detection of banned MBM in feedstuffs by reinforcement of the monitoring of feeds.     

Estimation of the stability of skeletal muscle myoglobin of chilled pork treated with brine activated by low-frequency high-intensity ultrasound

We studied the effect of ultrasonic activation of brine (3%) during salting on the degree of stability of colour parameters of pork with normal (NOR) and abnormal course of autolysis in the CIE Lab colour space. The mechanism of stabilisation of the colour of meat is attributed to donor–acceptor bonds of metmyoglobin (MetMb). The accumulation of excessive number of free electrons in the medium are capable of activating MetMb. This reduces the activity of meat, when the native participants of the metmyoglobin reductase system and their own antioxidant systems of meat are depleted.Based on the additive calculation of deviations (increase / decrease) by the coordinates L*, a*, b* in the CIE Lab system, and the total colour difference (ΔE) in control and experimental samples, recommendations were developed. To optimize the colour characteristics of all types of meat, both on the surface and in the thickness of the meat, the preliminary activation of a 3% brine in a low-frequency submersible ultrasonic unit is recommended. Moreover, preliminary cavitation activation of a 3% is more preferable to stabilise the colour of PSE – meat (pale, soft, exudative (watery),) brine in a flow-through installation.     

Evaluation of the influence of proline, hydroxyproline or pyrrolidine in the presence of sodium nitrite on N-nitrosamine formation when heating cured meat

N-nitrosamines are meant to be probable or possible carcinogenic components, possibly formed out of a reaction between nitrite and N-containing substances such as amino acids and secondary amines. Nitrite is often used for processing meat products because of its colouring and antimicrobial properties.During this experimental setup, the influence of proline, hydroxyproline or pyrrolidine on N-nitrosamine formation in meat samples was evaluated. The N-nitrosamines concentrations were measured with gas chromatography-thermal energy analyzer. Only the concentrations of N-nitrosodimethylamine and N-nitrosopyrrolidine were found above the limit of detection in a number of tested experimental conditions. The concentration of these two N-nitrosamines was modelled as a function of temperature and nitrite concentration for different situations (presence or absence of added natural N-containing meat components). It could be concluded that proline and pyrrolidine promoted the formation of N-nitrosopyrrolidine. It could also be confirmed that the higher the temperature of the meat processing procedure and the higher the sodium nitrite amounts added, the higher were the yields of the respective N-nitrosamines.     

Estimating Aluminum leaching from Aluminum cook wares in different meat extracts and milk

A method of estimating Al leaching from Al cook ware in some meat extracts and liquid milk was investigated. In the present work four kinds of Al cook wares from four countries were chosen from the local market. Extracts of boiled meat (lamb, chicken, and fish) were used to make 10–50% (w/v) concentrations. In addition liquid fresh milk and long life milk were diluted to make 10–50% (v/v) concentrations. Weight loss (WL) method, atomic absorption, polarization method, and surface study were applied. The “estimated” Al intake per person from WL in 30% meat extract and milk range from 8.16 to 12.75 mg/h with fish extract having the highest leaching and chicken extract having the lowest leaching. Atomic absorption gave comparable results to that of WL. Comparing the present results with the Provisional Tolerance Weekly Intake of Al approved by the FDA/WHO shows that Al leaching from Al cook wares may add high doses of Al into the diet.     

Sulfonate group-modified FePtCu nanoparticles as a selective probe for LDI-MS analysis of oligopeptides from a peptide mixture and human serum proteins

The availability of robust methods for the species-specific detection of meat and bone meal (MBM) in compound feedingstuffs is an important prerequisite to enforce current and upcoming European legislation on the use of processed animal proteins in animal nutrition. Among possible methods, those based on DNA turned out to be a reliable tool for this aim, since DNA is a quite thermostable molecule able to resist severe heat treatments applied in the manufacturing of animal meals. The application of such methods by control laboratories implies that the method has been validated including an assessment of its robustness. Successful transferability between laboratories is considered an important robustness criterion of the method. However, corresponding guidelines regarding the design of such a study relevant to this field are missing. Here, we demonstrate the feasibility of an alternative concept that was applied to check for the transferability of a qualitative assay for the detection of banned MBM in feedingstuffs at trace level based on real-time PCR. The concept was based on an experimental nested design applying analysis of variance (ANOVA) that was conducted independently in two laboratories and which allows for establishing major factors influencing the result of analysis. Statistical assessment of the results confirmed the importance of the DNA extraction/purification step utilised, whereas the PCR step turned out to be a minor factor regarding the overall variability of the results. Furthermore, blind samples comprised of compound feed adulterated with MBM at 0.1 % and blank compound feed were correctly classified as "positive" or "negative" samples, thus confirming fitness of purpose for the method. This approach can be of interest for other research groups working in the development of real-time PCR methods and in their use by control laboratories. Nested design of the study to check for the transferability of the real-time PCR method     

猪肉中二甲胺四环素的反相-高效液相色谱测定

建立了一种测定猪肉中二甲胺四环素的反相-高效液相色谱(RP-HPLC)分析方法.目标物经EDTA-Mcllvaine提取,用Oasis HLB固相萃取小柱净化,流动相为甲醇-咪唑缓冲溶液,体积比20:80,荧光检测器检测,外标法定量.平均添加回收率在89.6％～94.0％之间,相对标准偏差(RSD)在3.1％～6.5％...     

Determination of volatile nitrosamines in meat products by microwave-assisted extraction and dispersive liquid-liquid microextraction coupled to gas chromatography-mass spectrometry

Microwave-assisted extraction (MAE) and dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-mass spectrometry (GC-MS) were evaluated for use in the extraction and preconcentration of volatile nitrosamines in meat products. Parameters affecting MAE, such as the extraction solvent used, and DLLME, including the nature and volume of the extracting and disperser solvents, extraction time, salt addition and centrifugation time, were optimized. In the MAE method, 0.25g of sample mass was extracted in 10mL NaOH (0.05M) in a closed-vessel system. For DLLME, 1.5mL of methanol (disperser solvent) containing 20μL of carbon tetrachloride (extraction solvent) was rapidly injected by syringe into 5mL of the sample extract solution (previously adjusted to pH 6), thereby forming a cloudy solution. Phase separation was performed by centrifugation, and a volume of 3μL of the sedimented phase was analyzed by GC-MS. The enrichment factors provided by DLLME varied from 220 to 342 for N-nitrosodiethylamine and N-nitrosopiperidine, respectively. The matrix effect was evaluated for different samples, and it was concluded that sample quantification can be carried out by aqueous calibration. Under the optimized conditions, detection limits ranged from 0.003 to 0.014ngmL(-1) for NPIP and NMEA, respectively (0.12-0.56ngg(-1) in the meat products).     

Flow injection on-line ultrasound-assisted extraction of iron in meat samples coupled to a flame atomic absorption spectrometric system

Iron was extracted on-line from solid meat samples by a simple and rapid continuous ultrasound-assisted extraction system (CUES). The CUES is connected to a flow injection manifold, which allows the on-line flame atomic absorption spectrometric determination of iron. A Plakett–Burman design was used for the optimisation of the CUES. The method achieved a total sampling frequency of 11 samples per hour with a relative standard deviation for the complete procedure of 0.4%. The detection limit was 0.6 g g^–1 (dry mass) for a sample amount of 30 mg. Accurate results were obtained by measuring the certified reference materials BCR-186 (pig kidney) and BCR-184 (bovine muscle). The analytical procedure was applied to different real meat samples with satisfactory results.     

Multiresidue analysis of sulfonamides in meat by supramolecular solvent microextraction,liquid chromatography and fluorescence detection and method validation according to the 2002/657/EC decision

A multiresidue method was described for determining eight sulfonamides, SAs (sulfadiazine, sulfamerazine, sulfamethoxypyridazine, sulfachloropyridazine, sulfadoxine, sulfamethoxazole, sulfadimethoxine and sulfaquinoxaline) in animal muscle tissues (pork, chicken, turkey, lamb and beef) at concentrations below the maximum residue limit (100 μg kg⁻¹) set by the European Commission. The method was based on the microextraction of SAs in 300-mg muscle samples with 1 mL of a supramolecular solvent made up of reverse micelles of decanoic acid (DeA) and posterior determination of SAs in the extract by LC/fluorescence detection, after in situ derivatization with fluorescamine. Recoveries were quantitative (98–109%) and matrix-independent, no concentration of the extracts was required, the microextraction took about 30 min and several samples could be simultaneously treated. Formation of multiple hydrogen bonds between the carboxylic groups of the solvent and the target SAs (hydrogen donor and acceptor sum between 9 and 11) were considered as the major forces driving microextraction. The method was validated according to the European Union regulation 2002/657/EC. Analytical performance in terms of linearity, selectivity, trueness, precision, stability of SAs, decision limit and detection capability were determined. Quantitation limits for the different SAs ranged between 12 μg kg⁻¹ and 44 μg kg⁻¹, they being nearly independent of matrix composition. Repeatability and reproducibility, expressed as relative standard deviation, were in the ranges 1.8–3.6% and 3.3–6.1%. The results of the validation process proved that the method is suitable for determining sulfonamide residues in surveillance programs.