水产品及其制品中大西洋鲑鱼实时荧光PCR检测方法建立

目的:三文鱼营养丰富,含有ω-3不饱和脂肪酸,深受大众喜爱,但存在产品标签标示错误、掺伪掺假、以次充好的现象。目前采用检测标准SN/T 3589.7—2013《出口食品中常见鱼类及其制品的鉴伪方法第七部》对鲑鱼成分检测,无法区分大西洋鲑鱼和其他种类的三文鱼。因此有必要建立一种用于区分上述几种三文鱼的快速检测方法。方法:研究建立了针对大西洋鲑鱼成分鉴定的实时荧光PCR法,并对该方法的特异性和检出限进行了验证。结果:该方法具有良好的特异性,检出限为1%(w/w)。结论:可应用于大西洋鲑鱼的日常检测,为监管部门提供技术支持。     

Synchronous fluorescence and multivariate classification analysis as a screening tool for determining Sudan I dye in culinary spices

Spices are a globally traded commodity which has been found to be adulterated with forbidden Sudan dyes. This work proposes a screening method for determining the adulteration of paprika varieties (mild, hot and smoked) with Sudan I dye, based on constant-wavelength synchronous fluorescence spectroscopy with multivariate classification. Different wavelength-intervals (Δλ) were evaluated. Classification models were built with Partial Least Squares-Discriminant Analysis (PLS-DA) at two Sudan I dye concentration levels (1 and 5 mg L⁻¹) and they were tested with samples at a lower level (0.5 mg L⁻¹). Classification results were quite satisfactory when a strategy based on first-derivative spectra was used for improving classification results. Δλ = 60 nm was chosen as the optimum wavelength interval giving a 100% of sensitivity and specificity. These results are promising because the risk of assigning adulterated samples as safe to be consumed is highly minimized. The proposed method is feasible, rapid and simple taking advantage of Sudan I fluorescence phenomena in a direct way.     

一种基于双重ddPCR的肉制品中牛源性成分的定量分析方法

将低价肉类原料掺入高价肉制品是一种典型的肉类掺假方式，为准确鉴别牛肉及牛肉制品的真伪，建立一种基于双重微滴式数字聚合酶链式反应（droplet digital polymerase chain reaction，ddPCR）的牛源性成分定量分析方法。通过对14 种动物的Beta-actin单拷贝基因序列进行分析，设计牛源性特异性引物、探针与动物源性成分通用引物、探针并建立双重ddPCR体系，推导出牛源性成分质量与其特异性扩增拷贝数之间的计算公式，对牛源性成分进行定量检测该方法得到牛源性成分质量M牛（mg）与其特异性扩增拷贝数浓度C牛（copies/μL）之间的计算公式为M牛=0.033C牛＋2.37，对已知牛肉质量的混合肉样品与不同部位的牛肉样品进行检测，结果显示牛肉定量检测值与实际质量基本一致。同时拷贝数相对含量可辅助判断肉制品中是否存在非牛源性的其他动物源性成分掺假。对市售样品的检测发现，存在目标肉样含量不达标以及不同程度的动物源性成分等掺假现象，说明该检测方法可为牛肉制品掺假量化判定和混合源性产品分级鉴定提供技术支撑。     

Determination of chlorine ions in raw milk by pulsed amperometric detection in a flow injection system

Chloride ion concentration in milk was determined by pulsed amperometric detection in a flow injection system. Results showed that the Au electrode lost 3 electrons at 1.10 V and formed chloroaurate ions (AuCl₄^?) by combining with chloride ions, after which AuCl₄^? was partly reduced to Au at 0.6 V. Based on the electrochemical process, a triple waveform with detection potential of 1.15 V, detection time of 150 ms, oxidation potential of 1.4 V, oxidation time of 550 ms, reduction potential of 0 V, and reduction time of 400 ms was applied to the Au electrode for detecting chloride ion concentration in milk. The approach is rapid and automatic and features a detection limit of 0.005 g/L. The relative standard deviation obtained by 60 repetitive injections reached 1.48% at 2 g/L of NaCl. The method developed using the Au electrode without modification was used to analyze the chloride ion concentration in raw milk without preprocessing. The method showed good agreement with potentiometric titration.     

超高效液相色谱-串联质谱法检测多基质保健食品中27种非法添加降糖类化合物

目的 建立超高效液相色谱-三重四极杆质谱检测保健食品中27种降糖类非法添加化合物的方法。方法 4种不同基质保健品样品经甲醇超声提取后，采用Waters XBridge Amide色谱柱 (2.1 mm×100 mm, 3.5 μm)，以5 mmol/L甲酸铵水溶液和5 mmol/L甲酸铵乙腈作为流动相进行梯度洗脱，分离苯乙双胍、二甲双胍、伏格列波糖、阿卡波糖、丁二胍共5种化合物；采用Waters CORTECS T3色谱柱 (2.1 mm×100 mm, 2.7 μm)，以0.1%甲酸水溶液和乙腈作为流动相进行梯度洗脱，分离维达列汀、罗格列酮、西他列汀、吡格列酮等22种化合物；环格列酮采用电喷雾离子化负离子模式，其余26种化合物采用电喷雾离子化正离子模式，以多反应监测方式进行检测。结果 伏格列波糖、阿卡波糖、达格列净、卡格列净、曲格列酮、环格列酮6种化合物在250 ng/mL~12.5 μg/mL范围内线性关系良好，其余21种化合物在5~250 ng/mL范围内线性关系良好，线性相关系数r均大于0.99；27种化合物检出限(limit of detection, LOD)范围为1~1000 μg/kg；在低、中、高三水平加标，其回收率均在70%~120%范围内，相对标准偏差(relative standard deviation, RSD)为0.7%~10% (n=6)。结论 本方法快速、灵敏、高效、准确，适用范围广，可用于保健食品中降糖类化合物的高通量检测。     

Implication of palynological techniques for the authentication of adulterated drugs traded with the same name in different herbal markets of district Lahore,Pakistan

Adulteration in traded medicinal plants is a significant issue nowadays and use of these adulterated medicinal plants can impose harmful impact to end user. However, this problem can be overcome by ensuring the identification of traded medicinal plants which are used in making different herbal medicines. In this regard, palynological markers are considered to be an important taxonomic tool in the identification of original medicinal plant from its adulterant. Hence this study attempted to provide particular reliable palynological markers for distinguishing selected medicinal plants from their adulterants, that is, Cinnamomum verum versus Canella winterana, Cinnamomum tamala versus Cinnamomum obtusifolium, Gymnema sylvestre versus Gymnema lactiferum, Artemisia maritima versus Artemisia absinthium, Achillea millefolium versus Adhatoda vasaka, Sphaeranthus indicus versus Sphaeranthus africanus, Averrhoa carambola versus Butea monosperma, and Morus nigra versus Morus alba. Results demonstrated great variations in multiple palynological characters between original medicinal plant and its adulterant such as in pollen size, shape, colpi length, exine, intine thickness, and fertility. In equatorial view, circular to spheroidal shape of pollen was found in A. millefolium while oblate shape was observed in A. vasaka. Similarly B. monosperma pollen was 34 μm, whereas pollen of its adulterant A. carambola was 21 μm. Moreover, colpi length of A. maritima was 11.8 μm, whereas 4.5 μm in A. absinthium. Hence it can be concluded that palynological characters are commendably helpful in identification of genuine medicinal plant from its adulterant.     

Use of MALDI-TOF MS technology to evaluate adulteration of small ruminant milk with raw bovine milk

Detection of adulteration of small ruminant milk is very important for health and commercial reasons. New analytical and cost-effective methods need to be developed to detect new adulteration practices. In this work, we aimed to explore the ability of the MALDI-TOF mass spectrometry to detect bovine milk in caprine and ovine milk using samples from 18 dairy farms. Different levels of adulteration (0.5, 1, 5, 10, 20, 40, 60, and 80%) were analyzed during the lactation period of goat and sheep (in May, from 60 to 90 d in milk, and in August, from 150 to 180 d in milk). Two different ranges of peptide-protein spectra (500–4,000 Da; 4–20 kDa) were used to establish a calibration model for predicting the concentration of adulterant using partial least squares and generalized linear model with lasso regularization. The low molecular weight part of the spectra together with the generalized linear model with lasso regularization regression model appeared to have greater potential for our aim of detection of adulteration of small ruminants' milk. The subsequent prediction model was able to predict the concentration of bovine milk in caprine milk with a root mean square error of 11.4 and 17.0% in ovine milk. The results offer compelling evidence that MALDI-TOF can detect the adulteration of small ruminants' milk. However, the method is severely limited by (1) the complexity of the milk proteome resulting from the adulteration technique, (2) the potential degradation of thermolabile proteins, and (3) the genetic variability of tested samples. Additionally, the root mean square error of prediction based only on one individual sample adulteration series can drop down to 6.34% for quantification of adulterated caprine milk and 6.28% for adulterated ovine milk for the full set of concentrations or down to 2.33 and 4.00%, respectively, if we restrict only to low concentrations of adulteration (0, 0.5, 1, 5, 10%).     

Duplex real-time PCR assay for the simultaneous determination of the roe deer (Capreolus capreolus) and deer (sum of fallow deer,red deer and sika deer) content in game meat products

Due to the high price of game meat, food producers may be tempted to adulterate their products with cheaper meat. This paper presents a duplex real-time PCR assay which allows the simultaneous determination of the content of roe deer (Capreolus capreolus) and deer* (the sum of fallow deer (Dama dama), red deer (Cervus elaphus) and sika deer (Cervus nippon)) in food products to detect food adulteration. Relative quantification is carried out by using a reference (“all meat”) PCR assay based on the myostatin gene. The quantification approach was validated by analyzing binary meat mixtures with pork, “all game” meat mixtures containing each of the four game species in pork and a model game sausage. Compared to singleplex assays the duplex assay is time and cost saving. Thus, it is highly applicable to routine analysis in order to verify the authenticity of game meat products.