Application of DNA Fingerprinting Technique in Adulterated Food Identification and Traceability

To further improvc the application of DNA fingerprinting technique in adulterated food identification and traceability, the paper briefly introduced the ap- plication of common DNA fingerprinting techniques, such as species-specific PCR, RAPD, AFLP, ISSR, SSR and SNP in adulterated food identification and traceability.

Application of principal component-radial basis function neural networks (PC-RBFNN) for the detection of water-adulterated bayberry juice by near-infrared spectroscopy

Near-infrared (NIR) spectroscopy combined with chemometrics techniques was used to classify the pure bayberry juice and the one adulterated with 10% (w/w) and 20% (w/w) water. Principal component analysis (PCA) was applied to reduce the dimensions of spectral data, give information regarding a potential capability of separation of objects, and provide principal component (PC) scores for radial basis function neural networks (RBFNN). RBFNN was used to detect bayberry juice adulterant. Multiplicative scatter correction (MSC) and standard normal variate (SNV) transformation were used to preprocess spectra. The results demonstrate that PC-RBFNN with optimum parameters can separate pure bayberry juice samples from water-adulterated bayberry at a recognition rate of 97.62%, but cannot clearly detect water levels in the adulterated bayberry juice. We conclude that NIR technology can be successfully applied to detect water-adulterated bayberry juice.

Analysis of Peanut Oil Adulterated with Other Edible Oils by Spectrophotometry

Since peanut oil（PO） is more expensive than other seed oils, some PO is adulterated with other cheap seed oils, such as soybean oil, palm olein, cottonseed oil, corn oil and rapeseed oil. The conventional method for deter mining whether PO was adulterated is to detect the freezing point of oils. The proposed method for the determination of adulterants in PO was based on monitoring the change of absorbance when the sample was refrigerated. A special spectrophotometer was developed. A total of 10 kinds of POs from different suppliers were chosen and adulterated with other seed oils at the volume fraction levels ranging from 5% to 30%. A total of 150 samples were analyzed by the proposed method and the results were satisfactory.

Detection of Glucomacropeptide in Raw Milk Adulterated with Cheese Whey in Ecuador

Milk is one of the products that can be adulterated in many ways affecting the quality of this and its derivatives. Glucomacropeptide (GMP) is a protein that is found only in the whey from the production of fresh cheese, enzymatically obtained from the coagulation of casein and which is commonly used to adulterate fresh or powdered milk. The aim of this study was to determine the adulteration of milk with cheese whey thought a molecular approach, where the glucomacropeptide was collected by sequential precipitation with trichloroacetic acid (ATC) and detected by polyacrylamidododecylsulfate gel electrophoresis (PAGE-SDS), using samples of fresh milk, intentionally adulterated with serum in the proportion of 0%, 1%, 5%, 10% and 15%. The results obtained showed that the detection of glucomacropeptide by electrophoresis was positive in all samples of adulterated milk, evidencing a band of 20.9 kDa in the reading, corresponding to the molecular weight of the GMP, showing that the technique used determines the adulteration in the milk, in a specific and sensitive way, also shows that in the evaluation of physical-chemical and microbiological parameters of milk, there are no significant differences between treatments, except for the pH that tends to decrease as the percentage of serum in the milk increases.

Quantification of the adulteration concentration of palm kernel oil in virgin coconut oil using near-infrared hyperspectral imaging

The adulteration concentration of palm kernel oil(PKO)in virgin coconut oil(VCO)was quantified using near-infrared(NIR)hyperspectral imaging.Nowadays,some VCO is adulterated with lower-priced PKO to reduce production costs,which diminishes the quality of the VCO.This study used NIR hyperspectral imaging in the wavelength region 900-1,650 nm to create a quantitative model for the detection of PKO contaminants(0-100%)in VCO and to develop predictive mapping.The prediction equation for the adulteration of VCO with PKO was constructed using the partial least squares regression method.The best predictive model was pre-processed using the standard normal variate method,and the coefficient of determination of prediction was 0.991,the root mean square error of prediction was 2.93%,and the residual prediction deviation was 10.37.The results showed that this model could be applied for quantifying the adulteration concentration of PKO in VCO.The prediction adulteration concentration mapping of VCO with PKO was created from a calibration model that showed the color level according to the adulteration concentration in the range of 0-100%.NIR hyperspectral imaging could be clearly used to quantify the adulteration of VCO with a color level map that provides a quick,accurate,and non-destructive detection method.

Adulteration detection of plant protein beverages by UPLC-MS/MS based on signature peptide of allergen

Plant protein beverage adulteration occurs frequently,which may cause health problems for consumers due to the hidden allergens.Hence,a novel method was developed for authentication by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Almond,peanut,walnut and soybean were hydrolyzed,followed by separation by NanoLC-Triple TOF MS.The obtained fingerprints were identified by ProteinPilotTM combined with Uniprot,and 16 signature peptides were selected.Afterwards,plant protein beverages treated by trypsin hydrolysis were analyzed with UPLC-MS/MS.This method showed a good linear relationship with R2>0.99403.The limit of quantification(LOQ)were 0.015,0.01,0.5 and 0.05 g/L for almond,peanut,walnut and soybean,respectively.Mean recoveries ranged from 84.77%to 110.44%with RSDs<15%.The developed method was successfully applied to the adulteration detection of 31 plant protein beverages to reveal adulteration and false labeling.Conclusively,this method could provide technical support for authentication of plant protein beverages to protect the rights and health of consumers.

Analysis of an Adulterated Herbal Medicinal Product Using Ultra-Performance Liquid Chromatography Coupled with QTOF Mass Spectrometry

The reports of severe adverse effects and fatalities associated with herbal medicinal products adulterated with synthetic compounds have raised global concerns.The objective of this study is to analyze one commercial herbal medicinal product suspected to be adulterated with synthetic drugs in order to identify potential adulterants,to verify if the product contained the herbs listed as ingredients in label claim and to determine quality consistency among different batches of the product.Analyses of suspected product obtained from seven different batches were performed using ultra performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) with multiple data processing tools and multivariate analyses.In addition,23 individual powdered herbs(12 as per label claim and 11 suspected herbs),11 marker compounds of the labeled herbs and five suspected synthetic drugs as adulterants were also concurrently analyzed to have clear understanding of product composition.Based on our analysis,the major ingredients of studied product were found to be 5 synthetic compounds:caffeine,chlorphenamine,piroxicam,betamethasone and oxethazaine.Three of them have been found to exceed their recommended doses.From the herbal composition analysis,Gan Cao(Glycyrrhizae radix et rhizoma) was found to be the main ingredient,which is not among the claimed 12 herbs that were supposed to be in the product.Other herbs detected as minor ingredients were Mu Gua(Chaenomelis fructus),Dang Gui(Angelicae sinensis radix),and Huang Qi(Astragali radix),which are among the 12 herbs that were supposed to be in the product.Based on our results we demonstrated that UPLC-QTOF MS is an effective and versatile tool for the analysis of herbal medicinal products.It is highly desirable to have a streamlined process with automatic workflow and fit-forpurpose database to increase efficiency and productivity of sample analysis.Results of this work also highlight the need for the better quality control and regulatory measures to protect consumers from the potentially harmful effects of such adulterated products.

Drug adulteration analysis based on complexation with cyclodextrin and metal ions using ion mobility spectrometry

Drug adulteration and contamination are serious threats to human health therefore,their accurate monitoring is very important.Allopurinol(Alp)and theophylline(Thp)are commonly used drugs for the treatment of gout and bronchitis,while their isomers hypoxanthine(Hyt)and theobromine(Thm)have no effect and affect the efficacy of the drug.In this work,the drug isomers of Alp/Hyt and Thp/Thm are simply mixed withα-,β-,γ-cyclodextrin(CD)and metal ions and separated using trapped ion mobility spectrometry-mass spectrometry(TIMS-MS).TIMS-MS results showed that Alp/Hyt and Thp/Thm isomers could interact with CD and metal ions and form corresponding binary or ternary complexes to achieve their TIMS separation.Different metal ions and CDs showed different separation effect for the isomers,among which Alp and Hyt could be successfully distinguished from the complexes of[Alp/Hyt+γ-CD+Cu–H]^(+)with separation resolution(RP–P)of 1.51;whereas Thp and Thm could be baseline separated by[Thp/Thm+γ-CD+Ca–H]^(+)with RP–P of 1.96.Besides,chemical calculations revealed that the complexes were in the inclusion forms,and microscopic interactions were somewhat different,making their mobility separation.Moreover,relative and absolute quantification was investigated with an internal standard to determine the precise isomers content,and good linearity(R^(2)>0.99)was obtained.Finally,the method was applied for the adulteration detection where different drugs and urine were analyzed.In addition,due to the advantages of fast speed,simple operation,high sensitivity,and no chromatographic separation required,the proposed method provides an effective strategy for the drug adulteration detection of isomers.

Validation of the Methods for Detection the Non-Milk Fat in a Mixture of Milk Fat and Palm Oil

Milk fat contains a variety of nutritive and health-promoting compounds that guard against some disease. In the current system of global competition, when the quality of milk and milk products is not an option but rather a requirement, therefore, determining the purity of milk fat is critical. This study aims to validate analytical methods for detecting palm oil in a mixture of milk fat and palm oil. Methods of this study was involved detection of non-milk fat in fat blinders by determining the saponification value, iodine number, refractive index, butyro refractometer reading, Gas chromatography, Reverse Phase High-performance liquid chromatography, and Fourier transforms Infrared. The results of this study revealed that the saponification value, Iodine number, refractive index, and Butyro Reading could be used to detect the addition of palm oil by a level of 10% - 20% or more to the milk. The level of some fatty acids in the milk as determined by GC, such as myristic acid (C14:0), palmitic acid (C16:0), and stearic acid (C18:0), is correlated well with the level of adding palm oil to milk fat. The determination of cholesterol and β-sito-sterol content by RP-HPLC could be used for the detection of the addition of palm oil to milk fat. The spectrum behavior produced by FTIR spectroscopy in this adulterated sample is almost the same, so this technique could not be used to detect the palm oil in milk fat.

Advances in Research of Identification of Cnidii Fructus and Its Adulterants

In this paper,the traditional identification and modern identification technology of Cnidii Fructus and its adulterant were reviewed,and their advantages,disadvantages and practicability were summarized,to provide a reference for the rapid and accurate identification and quality evaluation of Cnidii Fructus.

Quality analysis of commercial samples of Ziziphi spinosae semen(suanzaoren) by means of chromatographic fingerprinting assisted by principal component analysis

Due to the scarcity of resources of Ziziphi spinosae semen （ZSS）, many inferior goods and even adulterants are generally found in medicine markets. To strengthen the quality control, HPLC fingerprint common pattern established in this paper showed three main bioactive compounds in one chromatogram simultaneously. Principal component analysis based on DAD signals could discriminate adulterants and inferiorities. Principal component analysis indicated that all samples could be mainly regrouped into two main clusters according to the first principal component （PC1, redefined as Vicenin II） and the second principal component （PC2, redefined as zizyphusine）. PC1 and PC2 could explain 91.42%of the variance. Content of zizyphusine fluctuated more greatly than that of spinosin, and this result was also confirmed by the HPTLC result. Samples with low content of jujubosides and two common adulterants could not be used equivalently with authenticated ones in clinic, while one reference standard extract could substitute the crude drug in pharmaceutical production. Giving special consideration to the well-known bioactive saponins but with low response by end absorption, a fast and cheap HPTLC method for quality control of ZSS was developed and the result obtained was commensurate well with that of HPLC analysis. Samples having similar fingerprints to HPTLC common pattern targeting at saponins could be regarded as authenticated ones. This work provided a faster and cheaper way for quality control of ZSS and laid foundation for establishing a more effective quality control method for ZSS.

Use of fluorometry for determination of skim milk powder adulteration in fresh milk

A FAST (fluorescence of advanced Maillard products and Soluble Tryptophan) method for identification of recon- stituted milk made from skim milk powder in the fresh milk was developed. Considering milk and skim milk powders variations from different seasons and countries, milk was collected from different dairy farms in different seasons and skim milk powders were collected from different countries to measure the Tryptophan (Trp), advanced Maillard products (AMP) fluorescence values. The results showed that there were differences (P<0.01) between raw and reconstituted milk. The plot of values in each mixed level of raw and reconstituted milk had a correlation coefficient >0.97. The FAST method is a simple, rapid, low-cost and sensitive method enabling the detection of 5% reconstituted milk in fresh milk. The measurement of the Trp, AMP fluorescence values and calculation of the FAST index is a suitable method for large-scale monitoring of fresh milk samples.

Identification of the varietal origin of processed loose-leaf tea based on analysis of a single leaf by SNP nanofluidic array

Tea is an important cash crop, representing a $40 billion-a-year global market. Differentiation of the tea market has resulted in increasing demand for tea products that are sustainably and responsibly produced. Tea authentication is important because of growing concerns about fraud involving premium tea products. Analytical technologies are needed for protection and value enhancement of high-quality brands. For loose-leaf teas, the challenge is that the authentication needs to be established on the basis of a single leaf, so that the products can be traced back to the original varieties. A new generation of molecular markers offers an ideal solution for authentication of processed agricultural products. Using a nanofluidic array to identify variant SNP sequences, we tested genetic identities using DNA extracted from single leaves of 14 processed commercial tea products. Based on the profiles of 60 SNP markers, the genetic identity of each tea sample was unambiguously identified by multilocus matching and ordination analysis. Results for repeated samples of multiple tea leaves from the same products(using three independent DNA extractions) showed 100% concordance, showing that the nanofluidic system is a reliable platform for generating tea DNA fingerprints with high accuracy. The method worked well on green, oolong, and black teas, and can handle a large number of samples in a short period of time. It is robust and cost-effective, thus showing high potential for practical application in the value chain of the tea industry.

Effects of Doped Elements on Electrochemical Performance of Ni(OH)_2 Materials

High energy ball milling （HEBM） method was applied to synthesize Ni （OH）2 with different doped elements sub-stitution for Ni^2＋. The morphology, structure and electrochemical behavior of prepared powders were studied. The re-suits reveal that all the synthesized Ni（OH）2 particles were in sub-micron sizes and greatly agglomerated. Co-, Mg-,Fe- or Mn-doped Ni （OH） 2 was of β-phase with 0.400-0.500 nm crystal interlayer distance, while A1- and Zn-doped products displayed a-phase with larger crystal interlayer spaces. The electrochemical mechanisms of synthe-sized Ni（OH）2 electrodes were discussed by EIS spectra. The specific capacity of Co-doped Ni （OH）2 is 245 mA·h · g^-1, i. e. , 60 mA· h · g^-1 higher than that of Al-doped electrode, which has the highest discharging plat-form of a mid-voltage of 1.30 V.

Rapid analysis of dyed safflowers by color objectification and pattern recognition methods

Objective:Rapid discrimination of three classes of safflowers,dyed safflower,adulterated safflower,and pure safflower using computer vision and image processing algorithms.Methods:A low cost computer vision system(CVS)was designed to measure the color of safflowers in the RGB(red,green,blue),L^*a^*b^*,and HSV(hue,saturation,vale)color spaces.The color moments in these three color spaces were extracted from the acquired images as color features of safflower.In addition,five kinds of pigments that are commonly used to dye safflowers were identified by high-performance liquid chromatography as a reference.Pattern recognition methods were investigated for rapid discrimination,including an unsupervised principal component analysis(PCA)algorithm and a supervised partial least squares discriminant analysis(PLS-DA)algorithm.Results:The mean error(e)between color values measured with the colorimeter and calculated with the CVS was 2.4%,with a high correlation coefficient(r)of 0.9905.This result indicated that the established CVS was reliable for color estimation of safflowers.The PLS-DA model,which had a total accuracy of 91.89%,outperformed the PCA model in classifying pure,adulterated,and dyed safflowers.Conclusion:The color objectification is a promising tool for rapid identification of dyed and adulterated safflowers.

Identifying camellia oil adulteration with selected vegetable oils by characteristic near-infrared spectral regions

In this paper,a methodology based on characteristic spectral bands of near infrared spectroscopy(1000-2500 nm)and multivariate analysis was proposed to identify camellia oil adulteration withvegetable oils,Sunflower,peanut and corn oils were selected to conduct the test.Pure camlia oiland that adulterated with varying concentrations(1-10%with the gradient of 1%,10-40%withthe gradient of 5%,40-100%with the gradient of 10%)of each type of the three vegetable oilswere prepared,respectively.For each type of adulterated oil,full-spectrum partial least squarespartial least squares(PLS)models and synergy interval partial least squares(SI-PLS)modelswere developed.Parameters of these models were optimized simultaneously by cross-validation,The SI-PLS models were proved to be better than the full-spectrum PLS models.In SI-PLSmodels,the correlation coefficients of predition set(Rp)were 0.9992,0.9998 and 0.9999 foradulteration with sunflower oil,peanut oiloil seperately;the corresponding root meansquare errors of prediction set(RMSEP).66nd 0.37.Furthermore,a new genericPLS model was built based on the chalselected from the intervals of thethree SI-PLS models to identify the oil adulterantsardless of the adultrated oil types.Themodel achieved with Rp=0.9988 and RMSEP==1.52,These results indicated that the charac-teristic near infrared spectral regions could determine the level of adulteration in the camllia oil.

Effect of Sodium Tri Polyphosphate (STPP) and Foreign Materials on the Quality of Giant Freshwater Prawn (<i>Macrobrachium rosenbergii</i>) under Ice Storage Condition

There are reports on the use of chemicals like sodium tri polyphosphate (STPP) and foreign materials like pearl tapioca (locally called ‘sagu’), jelly (litchi) to adulterate freshwater prawn (Macrobrachium rosenbergii) prior to freeze processing to increase their weight. Studies were, therefore, undertaken to determine the changes in product quality on the use of different concentrations of STPP, sagu and litchi under ice storage condition. Percent weight gain of prawn was 5.46, 18.87 and 23.50 when dipped in 2%, 4% and 6% STPP solution, respectively. In all cases maximum water uptake by prawn muscle was during the first 6 h with fastest weight gain with STPP solutions containing tap water compared to those of ice and tap water. Organoleptic quality of the STPP treated samples became brown and spongy after 8 h of dipping treatment under iced condition. Quality assessment studies conducted after injecting sagu and litchi in prawn muscle showed little or no difference with those of control samples during the first 6 h, which turned whitish and swollen with severe drip loss after 24 h of ice stored condition, indicating characteristics for easy identification of the injected shrimps by organoleptic method.

Identification of sea snake meat adulteration in meat products using PCR-RFLP of mitochondrial DNA

PCR-RFLP based technique for identification of sea snakes in Thai waters was achieved by developing species-specific markers.To distinguish between sea snake species,the PCR products of cytochrome b(Cyt b),12S and 16S rRNA were sequenced and cut with different restriction endonuclease,Alu I and Hinf I.Each enzyme generated different-sized fragments which specific to Cyt b of eight sea snake species.However,the identical pattern was found among Hydrophis group.This result could be resolved by using these enzymes 12S rRNA digestion.This technique was successfully applied to blood,shed skin,raw meat,cooked meat,sea snake-fish binary admixture,and sea snake-pork binary admixture.Hence,it could be applied for identification when sea snake meat adulteration in meat products and sold as meatballs to reduce production costs.Hopefully,this technique would improve sea snake species identification when morphological examination is no longer possible because the animals are already processed.This is very important to track when sea snake species are being hunted and also used to assess the conservation and management of the sea snakes in Thai waters,especially the Gulf of Thailand.

Rapid and Sensitive Identification of Horse and Donkey Meat in Iraqi Markets Using SSR and PCR-RFLP Based on Mitochondrial DNA Cytochrome B Gene

This study was carried out at the Genetic Engineering Lab, College of Agriculture, University of Basrah. DNA was extracted from samples containing meat of commercial products in the Basrah markets. The products subjected to analysis using polymerase chain reaction （PCR） with species specific repeat （SSR） and polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） techniques based on the sequence of the mitochondrial DNA cytochrome b gene （mtDNA cyt b gene）. Then digestion with the Alu l restriction enzyme to establish a differential diagnosis detects and discriminates between meat species and adulteration in the products. SSR primers were applied, has been detected amplification of the encoded gene product, generated 221 bp in some imported minced and canned meat samples. The results show that SSR analysis produced a pattern that allowed a direct identification of horse and donkey meats in some imported minced and canned meat samples （Hana, Monde and Bavaria）. The amplified 359 bp gene of mtDNA cyt b gene from samples in different product was cut using Alu 1 restriction enzyme resulting in restriction fragment length polymorphism （RFLP）. Alu 1 was used to distinguish between the animals meat that belong to the family or one species. The digestion of the PCR product showed differences between products. Where the fragment length generated were 74, 76 and 189 bp. It belonged to horse meat. The fraud was detected in Hana, Monde and Bavaria products available in Basrah markets showing the presence of horse meat in these products that labeled as beef meats 100%. This revealed mtDNA cyt b gene as highly conserved and consequently a good molecular marker for diagnostic studies. Thus, this technique can be applied to food authentication for the identification of different species of animals in food products.

Feasibility Study for Applying Spectral Imaging for Wheat Grain Authenticity Testing in Pasta

Authentication of pasta is currently determined using molecular biology-based techniques focusing on DNA as the target analyte. Whilst proven to be effective, these approaches can be criticised as being destructive, time consuming, and requiring specialist instrument training. Advances in the field of multispectral imaging (MSI) and hyperspectral imaging (HSI) have facilitated the development of compact imaging platforms with the capability to rapidly differentiate a range of materials (inclusive of grains and seeds) based on surface colour, texture and chemical composition. This preliminary investigation evaluated the applicability of spectral imaging for identification and quantitation of durum wheat grain samples in relation to pasta authenticity. MSI and HSI were capable of rapidly distinguishing between durum wheat and adulterant common wheat cultivars and assigning percentage adulteration levels characterised by low biases and good repeatability estimates. The results demonstrated the potential for spectral imaging based seed/grain adulteration testing to augment existing standard molecular approaches for food authenticity testing.