Synthesis of a novel composite imprinted material based on multiwalled carbon nanotubes as a selective melamine absorbent

A novel composite imprinted material, on the basis of a multiwalled carbon nanotube (CNT)-incorporated layer using melamine as a template, methacrylic acid as a functional monomer, and ethylene glycol dimethacrylate as a cross-linker, was synthesized by a surface imprinting technique. The imprinted/CNT sorbent was characterized by a scanning electron microscope (SEM). Adsorption dynamics and a Scatchard adsorption model were employed to evaluate the adsorption process. The results showed that the imprinted/CNT sorbent displayed a rapid dynamic adsorption and a high adsorption capacity of 79.9 μmol g(-1) toward melamine. Applied as a sorbent, the imprinted/CNT sorbent was used for the determination of melamine in a real sample by online solid-phase extraction-high-performance liquid chromatography (SPE-HPLC). An enrichment ratio of 563-fold, detection limit (S/N = 3) of 0.3 μg L(-1), and quantification limit of 4.5 μg L(-1) were achieved.     

线扫描式拉曼高光谱成像技术无损检测奶粉三聚氰胺

为了实现颗粒状样本的大面积无损快速检测,该研究结合拉曼光谱和高光谱技术搭建了一套线扫描式拉曼高光谱检测系统,对奶粉和三聚氰胺颗粒混合样本进行了检测研究。研究通过高斯窗平滑法和air PLS基线校正方法分别消除了拉曼光谱中的噪声信号和荧光背景,选取三聚氰胺主要特征峰(671.71 cm-1)处的单波段图像作为是否含有三聚氰胺颗粒的判断依据。研究首先对三聚氰胺产生的拉曼信号在奶粉颗粒中的穿透深度进行了检测,随后完成了10种不同浓度的三聚氰胺奶粉混合样本的拉曼高光谱采集,对特征单波段图像中各像素点的拉曼强度平均值进行一元线性分析,并对单波段图像进行二值化处理。结果显示,在三聚氰胺特征单波段图像中,感兴趣区域内所有像素点的拉曼强度平均值与三聚氰胺浓度之间线性度较高,其决定系数R2达到了0.995 4。在二值图像中,三聚氰胺颗粒的位置信息能够直观的展现。研究结果表明,拉曼高光谱成像技术具有快速、无损和大面积检测的特点,在实际应用中具有巨大潜力。     

Analytical method for the determination of cyromazine and melamine residues in soil using LC-UV and GC-MSD

A method is reported for the determination of cyromazine and melamine residues in soil. Soil samples are extracted twice via mechanical shaking, each time with 70% acetonitrile/30% 0.050 M ammomium carbonate for 30 min. An aliquot portion of the pooled extracts is subjected to strong cation exchange (SCX) purification on AG 50W-X4 resin. Final analysis is accomplished using liquid chromatography-ultraviolet (LC-UV) detection at a wavelength of 214 nm. Confirmatory analyses can be performed using gas chromatography-mass selective detection (GC-MSD) in the selected ion monitoring (SIM) mode. The limit of detection (LOD) is 2.5 ng injected and the limit of quantification (LOQ) is 10 ppb when using LC-UV for the analysis of N-cyclopropyl-1,3,5-triazine-2,4, 6-triamine (cyromazine) and 1,3,5-triazine-2,4,6-triamine (melamine). The LOD is 0.050 ng injected and the LOQ is 10 ppb when using GC-MSD for confirmatory analyses. The mean procedural recoveries were 97 and 95% and the standard deviations were 16 and 11% for cyromazine and melamine, respectively (n = 24), when using LC-UV. The mean procedural recoveries were 107 and 92% and the standard deviations were 9.9 and 16% for cyromazine and melamine, respectively (n = 29), when using GC-MSD. The method validation study was conducted under U.S. EPA FIFRA Good Laboratory Practice Guidelines 40 CFR 160. The method also passed an Independent Laboratory Validation (ILV) as per U.S. EPA FIFRA Subdivision N.     

Development of an immunochromatographic strip test for rapid detection of melamine in raw milk, milk products and animal feed

A simple, rapid and sensitive immunogold chromatographic strip test based on a monoclonal antibody was developed for the detection of melamine (MEL) residues in raw milk, milk products and animal feed. The limit of detection was estimated to be 0.05 μg/mL in raw milk, since the detection test line on the strip test completely disappeared at this concentration. The limit of detection was 2 μg/mL (or 2 μg/g) for milk drinks, yogurt, condensed milk, cheese, and animal feed and 1 μg/g for milk powder. Sample pretreatment was simple and rapid, and the results can be obtained within 3-10 min. A parallel analysis of MEL in 52 blind raw milk samples conducted by gas chromatography-mass spectrometry showed comparable results to those obtained from the strip test. The results demonstrate that the developed method is suitable for the onsite determination of MEL residues in a large number of samples.     

Selective determination of melamine in aqueous medium by molecularly imprinted solid phase extraction

A molecularly imprinted polymer able to recognize melamine in partially aqueous medium was synthesized using methacrylic acid as functional monomer and ethylene glycol dimethacrylate as cross-linking agent. The bound specificity and selectivity of the obtained material were verified by performing binding experiments with melamine and its structural analogue, 2,4,6-trimethoxy-1,3,5-triazine, respectively, using different aqueous binding media. Finally, the ability of MIP to selectively extract melamine from two real samples, a food supplement and a freeze-dried meat sample, was demonstrated.     

Detection of food additives by voltammetry at the liquid-liquid interface

Electrochemistry at the liquid-liquid interface enables the detection of nonredoxactive species with electroanalytical techniques. In this work, the electrochemical behavior of two food additives, aspartame and acesulfame K, was investigated. Both ions were found to undergo ion-transfer voltammetry at the liquid-liquid interface. Differential pulse voltammetry was used for the preparation of calibration curves over the concentration range of 30-350 microM with a detection limit of 30 microM. The standard addition method was applied to the determination of their concentrations in food and beverage samples such as sweeteners and sugar-free beverages. Selective electrochemically modulated liquid-liquid extraction of these species in both laboratory solutions and in beverage samples was also demonstrated. These results indicate the suitability of liquid-liquid electrochemistry as an analytical approach in food analysis.     

Efficient fluorescence energy transfer system between CdTe-doped silica nanoparticles and gold nanoparticles for turn-on fluorescence detection of melamine

We here report an efficient and enhanced fluorescence energy transfer system between confined quantum dots (QDs) by entrapping CdTe into the mesoporous silica shell (CdTe@SiO?) as donors and gold nanoparticles (AuNPs) as acceptors. At pH 6.50, the CdTe@SiO?-AuNPs assemblies coalesce to form larger clusters due to charge neutralization, leading to the fluorescence quenching of CdTe@SiO? as a result of energy transfer. As compared with the energy transfer system between unconfined CdTe and AuNPs, the maximum fluorescence quenching efficiency of the proposed system is improved by about 27.0%, and the quenching constant, K(sv), is increased by about 2.4-fold. The enhanced quenching effect largely turns off the fluorescence of CdTe@SiO? and provides an optimal "off-state" for sensitive "turn-on" assay. In the present study, upon addition of melamine, the weak fluorescence system of CdTe@SiO?-AuNPs is enhanced due to the strong interactions between the amino group of melamine and the gold nanoparticles via covalent bond, leading to the release of AuNPs from the surfaces of CdTe@SiO?; thus, its fluorescence is restored. A "turn-on" fluorimetric method for the detection of melamine is proposed based on the restored fluorescence of the system. Under the optimal conditions, the fluorescence enhanced efficiency shows a linear function against the melamine concentrations ranging from 7.5 × 10?? to 3.5 × 10?? M (i.e., 1.0-44 ppb). The analytical sensitivity is improved by about 50%, and the detection limit is decreased by 5.0-fold, as compared with the analytical results using the CdTe-AuNPs system. Moreover, the proposed method was successfully applied to the determination of melamine in real samples with excellent recoveries in the range from 97.4 to 104.1%. Such a fluorescence energy transfer system between confined QDs and AuNPs may pave a new way for designing chemo/biosensing.     

Novel application of square-wave adsorptive-stripping voltammetry for the determination of xanthohumol in spent hops

This paper reports the development of a novel electrochemical assay for xanthohumol (XN) by square-wave adsorptive-stripping voltammetry (SWAdSV) with a hanging mercury drop electrode. The method showed good repeatability (CV < 2%) and linearity (between 10 and 250 μg L(-1)), as well as suitable limits of detection (2.6 μg L(-1)) and quantification (8.8 μg L(-1)). The method was applied for the quantification of this compound in spent hops, and the results obtained were compared with the HPLC-UV method. XN contents determined by the SWAdSV method were 16 ± 1 and 100 ± 4 μg L(-1) for aqueous and methanolic extracts, respectively. The developed new methodology considerably reduces the analysis time, approximately from 25 min (HPLC-UV method) to 7 min, enabling a high sample throughput. In addition, the detection and quantification limits were approximately 5-fold lower than those obtained with the chromatographic method.     

高效液相色谱法同时测定土壤中环丙氨嗪和三聚氰胺

本试验研究建立了同时测定土壤中环丙氨嗪和三聚氰胺残留量的高效液相色谱法.红壤、潮土等5种土壤样品经氨水/甲醇 (5/95,v/v)超声提取3次,浓缩处理后上机检测.环丙氨嗪和三聚氰胺的标准曲线在0.1 ～ 15.0 μg/ml浓度范围内线性关系良好,绝对系数(R2)分别为1.0000和0.9998;在0.5 ～ 5 mg/kg添加范围内,环丙氨嗪和三聚氰胺在土壤中的平均回收率分别为87.2% ～ 101.1% 和 75.3% ～ 101.6%,变异系数分别为3.3% ～ 8.1%、1.6% ～ 9.9%,最低检测限分别为0.05 mg/kg、0.07 mg/kg.与国际上气相/液相色谱-质谱连用法相比,操作简单,经济方便易于普及.     

Diagnostic determination of melamine and related compounds in kidney tissue by liquid chromatography/tandem mass spectrometry

In 2007, it was determined that melamine, ammeline, ammelide, and cyanuric acid (abbreviated as MARC for melamine and related contaminants) had been added to wheat gluten and rice protein that were subsequently incorporated into pet food. The consumption of food tainted by MARC compounds was implicated in numerous instances of renal failure in cats and dogs. A method for the analysis of MARC compounds in kidney tissue using high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) has been developed. MARC analytes were extracted by homogenization of kidney tissue in 50/40/10 acetonitrile/water/diethylamine. The homogenate was centrifuged, and an aliquot of supernatant was diluted with acetonitrile, concentrated, and fortified with a stable isotope-labeled analogue of melamine. Analytes were detected using atmospheric pressure chemical ionization and multiple reaction monitoring. Quantitation of positive samples was performed using the internal standard method and five-point calibration curves ranging between 50 and 1000 ng/mL of each analyte. The method was validated by analysis of replicate kidney tissue samples fortified with the individual analytes and by analysis of kidney samples fortified with melamine cyanurate powder at two different concentrations. This method was successfully used for routine postmortem diagnosis of melamine toxicosis in animals. Melamine was also detected by this method in paraffin-embedded tissue from animals suspected to have died of melamine toxicosis.     

Determination of benzene in polypropylene food-packaging materials and food-contact paraffin waxes

An analytical procedure was developed for determination of benzene in polypropylene food packaging and was adapted for determination of benzene in commercial paraffin waxes intended for food-contact use. The polymer was dissolved in hexadecane at 150 degrees C. The wax was melted in an 80 degrees C oven. A simple helium-sparging apparatus was used to remove the volatile chemical from the polymer or wax. The contaminant was collected in methanol, distilled water was added, and the resulting solution was analyzed by headspace gas chromatography. The instrument was equipped with a 30 m fused silica open tubular capillary column and a photoionization detector. Average recoveries of benzene from polymer and paraffin wax at low parts-per-billion concentrations were 63 and 70%, respectively. Limits of detection and quantitation for analysis of polypropylene were 8 and 17 ppb, respectively; the limit of quantitation for analysis of paraffin wax was 2 ppb. In several commercial polypropylene products examined, benzene levels ranged from none detected to 426 ppb. In 3 commercial waxes examined, concentrations of 16-73 ppb benzene were determined. The presence of benzene was confirmed by gas chromatography/mass spectrometry.     

Development of gold nanoparticle-based rapid detection kit for melamine in milk products

A reliable and sensitive kit for the rapid detection of melamine (Mel) was developed. The kit is based on gold nanoparticle (Au NP) probe and includes a standard colorimetric card. The Au NPs were prepared by sodium borohydride reduction and characterized by transmission electron microscopy, which revealed particle sizes of approximately 5 nm. The performance of the kit in terms of aggregation kinetics, cross-reactivity, anti-interference, and sample pretreatment was investigated. The standard colorimetric card was then fabricated by chromatic aberration of a series of standard Mel-spiked milk reacts with the 5 nm Au NPs. The working range of the kit is 1-120 mg/L, and its performance is visibly more rapid and reliable by comparison with the standard colorimetric card. As low as 1 mg/L of Mel levels in milk can be determined, with the assay taking only about 10 min, including sample pretreatment. The kit can be stored for a year at room temperature. Samples were also detected by the kit, yielding results close to those obtained by high-performance liquid chromatography/mass spectrometry. Thus, the kit is applicable to qualitative and semiquantitative field detection, as well as naked-eye screening without the aid of any instrumentation.     

利用qPCR定量检测水体中猪源拟杆菌特异性生物标记的研究

以猪排泄物构建的污染水体为研究对象,通过寄主特异性引物识别水体中猪源拟杆菌16SrRNA基因的特异性基因标记（又称为特异性生物标记）,并根据特异性生物标记的定量检测,确定粪便拟杆菌的污染量,以明确水体受猪场废水污染的程度。在比较3种不同DNA提取方法的基础上,选择并改进CTAB法,建立了一种适合提取水样中猪排泄物总DNA的改良CTAB法。3对不同猪源寄主特异性引物的特异性验证结果表明,引物3（Bac32F／Bacl08R）的特异性好,检出下限在4．09E＋02~1．60E＋03拷贝数之间,更适用于水体中猪源拟杆菌属特异性生物标记的检测。不同混合污水中其他寄主来源的拟杆菌对猪源拟杆菌属特异性生物标记无影响,表明该方法可排除其他寄主来源拟杆菌的干扰,具有很好的特异性。     

Amperometric Biosensor Approaches for Quantification of Indole 3-Acetic Acid in Plant Stress Responses

Amperometric biosensors are known to be sensitive, reliable, and inexpensive instruments for biomolecular detection. Recently, self-referencing amperometric biosensors have been utilized to quantify the endogenous apoplastic flux of plant hormone indole 3-acetic acid (IAA) in vivo. There is still a significant need for sampling and testing methods to measure IAA concentrations in whole tissue samples. In the present study we used nanomaterial-modified platinum microelectrodes for the detection of IAA extracted from whole plant tissues. The key to the use of the nanomaterials was to enhance the surface area and thus the limit of detection for IAA. The nanoscale electrochemical interface was modified by the application of a layer of platinum black, followed by silanization overnight and a coating of multiwalled carbon nanotubes. Electroanalytical characterization of sensor performance was evaluated using both IAA standards and IAA extracted from corn plant samples, using a three-electrode scheme (reference, sensing, and auxillary). We compared tissue concentrations in water- and salt-stressed corn seedlings and compared these results to values measured using established enzyme-linked immunosorbent assay (ELISA) protocols. We found that the values obtained from both methods were comparable. The data obtained from the IAA sensor suggested that the electroanalytical biosensor approach was slightly more reliable and sensitive. Our results demonstrate a novel nanomaterial biosensor approach for IAA quantification in plant tissue extracts. The results of the stress study clearly indicated that root and shoot elongation and growth had significant positive correlation with IAA content in the raw plant tissue extracts. Water and salt stress both reduced root and shoot growth, which may be due to overaccumulation of IAA, resulting in inhibition of elongation. In comparison to established methods, such as high-performance liquid chromatography (HPLC) or ELISA, our approach is simple and inexpensive. Unlike HPLC, our approach does not require any elaborate sample purification, nor does it require antibody development as needed for ELISA. In summary, this electroanalytical biosensor method can be effectively utilized for simple, cheap, and reliable detection of IAA extracted from plant samples.     

Liquid chromatographic determination of melamine in beverages

A liquid chromatographic method is described for the determination of melamine in beverages. Melamine is separated by column chromatography using cation and anion exchange resin and determined by ion-pair liquid chromatography using an ODS column and a mixture of acetonitrile and 0.05M phosphate buffer (pH 3.0) containing 0.005M sodium 1-laurylsulfate (1 + 4, v/v) as mobile phase. Recoveries of melamine ranged between 90.3 +/- 7.8 and 102.1 +/- 5.6% at levels of 0.6 to 2.4 ppm in 4 kinds of beverages. The quantitation limit was 2.5 micrograms melamine in 50 mL beverage. The method was applied to the migration test of melamine from melamine-formaldehyde resin products to the beverages.     

Simultaneous detection of eight genetically modified maize lines using a combination of event- and construct-specific multiplex-PCR technique

To fulfill labeling and traceability requirement of genetically modified (GM) maize for trade and regulation, it is essential to develop an event-specific detection method for monitoring the presence of transgenes. In pursuit of this purpose, we systematically optimized and established a combined event- and construct-specific multiplex polymerase chain reaction (mPCR) technique for simultaneous detection of 8 GM maize lines. Altogether 9 sets of primers were designed, including six that were event-specific for Event176, Bt11, TC1507, NK603, MON863, and Mon810; two that were construct-specific for T25 and GA21, and one for an endogenous zein gene. The transgene in each GM maize line and the endogenous zein gene could be clearly detected and distinguished according to the different sizes of PCR amplicons. The limit of detection (LOD) was approximately 0.25% (v/v), although the detection can be as sensitive as 0.1% as demonstrated by the International Seed Testing Association (ISTA) proficiency test. This study further improves the current PCR-based detection method for GM maize. The method can be used in an easy, sensitive, and cost and time effective way for the identification and quality screening of a specific GM maize line.     

三聚氰胺对斜生栅藻的毒性研究

采用光照培养箱培养斜生栅藻的方法,以甘油为溶剂,研究了三聚氰胺对斜生栅藻的毒性效应。结果显示,当三聚氰胺的浓度在50、200 mg.L-1时,对斜生栅藻生长有促进作用,在750 mg.L-1时则表现出抑制效应,染毒7 d后对叶绿素的影响和对藻密度的影响一致,显示浓度-效应关系,即浓度越高抑制效应越明显。三聚氰胺对斜生栅藻超氧化物歧化酶（SOD）的影响表现为：当三聚氰胺浓度为200 mg.L-1时酶的活性达到峰值,然后开始下降,在750 mg.L-1 SOD的活性与对照相比减少了43.6%,三聚氰胺对斜生栅藻的脂质过氧化产物丙二醛（MDA）的影响和对可溶性糖的影响效应是一致的,随着浓度的增加均在减少。     

Polychlorinated biphenyls (PCBs) detection in food samples using an electrochemical immunosensor

In this work, a disposable electrochemical immunosensor, based on a competitive assay scheme, was applied to detect polychlorinated biphenyls (PCBs) in food. For this purpose, antibodies against PCBs were directly immobilized onto the carbon surface of a disposable screen-printed electrode. A competition between the PCBs present in the sample and a fixed concentration of an enzyme-labeled PCB was realized and evaluated by electrochemical detection. Alkaline phosphatase was used as the enzyme label, coupled with differential pulse voltammetry (DPV) as the electrochemical technique. The immunosensor was tested on aroclor mixture detection (1242 and 1248) and then on some typologies of food samples to evaluate the possible application for real sample analysis. Samples analyzed were from different matrixes, such as sheep milk, bovine adipose tissue, and bovine muscle. Results obtained were compared with the accredited results according to ISO 17025 methods for PCB detection (HRGC-LRMS) as a confirmatory analysis. Preliminary results show the possibility to use this device as a screening method in food sample analysis. The negligible matrix effect observed may lead to a simplified extraction procedure, and considerable time and consumable savings are the immediate benefits given by the proposed method.     

Square-wave voltammetric determination of acetaldehyde in povidone

A rapid, precise, and accurate method is described for the determination of acetaldehyde in povidone (polyvinyl pyrrolidone). The sample is dissolved in a basic aqueous electrolyte and analyzed using square-wave voltammetry. The proposed method allows for direct measurement of acetaldehyde under gentle conditions, thus avoiding loss of analyte due to volatility of acetaldehyde. The method is specific for acetaldehyde in the presence of other aldehydes and povidone process impurities.     

基于离子排斥色谱的饲料酸化剂中多种有机酸同步检测

为了规范饲用酸化剂的使用和管理,建立一种快速、准确的多种有机酸同步检测方法应需而生。该文以实现饲料酸化剂中多种有机酸的同步检测为目标,基于离子排斥色谱技术,对饲料酸化剂中多种有机酸(柠檬酸、酒石酸、苹果酸、乳酸、甲酸、乙酸、富马酸、丙酸和丁酸)的检测条件进行了优化研究。研究采用Bio-Rad Aminex HPX-87H(300mm×7.8 mmΦ,9μm)离子排斥色谱柱及二极管阵列检测器,以硫酸溶液作为流动相,比较了不同流动相浓度、不同柱温、不同流动相流速、不同检测波长等色谱条件对同步检测饲料酸化剂中9中有机酸的影响;以上述优选的条件建立分析方法,考察此方法的线性范围和检出限、回收率和精密度,并用实际样品进行了验证。研究结果表明:以0.025 mol/L硫酸溶液为流动相,流速0.70 m L/min,柱温30℃,DAD检测波长为210 nm,以保留时间结合待测物质的紫外特征吸收光谱进行定性分析,外标法进行定量分析。9种有机酸组分的质量浓度与其峰面积在一定范围内呈良好线性关系,决定系数0.999,9种有机酸组分在20 min内可以得到良好分离;方法检出限为0.01~2.4 mg/L,加标回收率为90.7%~101.1%,相对标准偏差为0.91%~3.33%。基于上述条件建立的检测方法能够满足饲料酸化剂中多种有机酸成分的同步定量分析。