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.

Molecular Identification of Chinese Materia Medica and Its Adulterants Using ITS2 and <i>psbA-trnH</i>Barcodes: A Case Study on Rhizoma Menispermi

Rhizoma Menispermi, derived from the rhizoma of Menispermum dauricum DC., is one of the most popular Chinese medicines. However Rhizoma Menispermi is often illegally mixed with other species in the herbal market, including Aristolochia mollissimae Hance, which is toxic to the kidneys and potentially carcinogenic. The use of DNA barcoding to authenticate herbs has improved the management and safety of traditional medicines. In this paper, 49 samples belonging to five species, including 34 samples of M. dauricum, from different locations and herb markets in China were collected and identified using DNA barcoding. The sequences of all 34 samples of Rhizoma Menispermi are highly consistent, with only one site variation in internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA and no variations in the psbA-trnH region. The intra-specific genetic distance is much smaller than inter-specific one. Phylogenetic analysis shows that both sequences allow the successful identification of all species. Nearest distance and BLAST1 methods for the ITS2 and psbA-trnH regions indicate 100% identification efficiency. Our research shows that DNA barcoding can effectively distinguish Rhizoma Menispermi from its adulterants from both commercial and original samples, which provides a new and reliable way to monitor commercial herbs and to manage the modern medicine market.

A Pharmacognostical Study on Ardisia gigantifolia and Its Adulterants

[Objectives]The research aimed to study the characteristics of Ardisia gigantifolia and its adulterants Embelia scandens and Mappianthus iodoides for identification. [Methods] Through the trait identification method,powder microscopic identification method,TLC,inclusions detection and content determination,A. gigantifolia and its adulterants were contrasted. [Results]The morphological and histological characteristics of A. gigantifolia,E. scandens and M. iodoides were different from each other. Containing water,total ash,acid insoluble ash and other inclusions also had difference; the total triterpenoid saponins were respectively 122. 90 mg/g of A. gigantifolia,147. 052 mg/g of E. scandens,and 63. 06 mg/g of M. iodoides. [Conclusions] These methods are accurate and reliable,which could be used for the identification of A. gigantifolia and its adulterants.

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.

DNA barcoding provides distinction between Radix Astragali and its adulterants

Based on variable nuclear and/or organellar DNA sequences among vastly divergent species as well as morphologically indistinguishable species, DNA barcoding is widely applicable in species identification, biodiversity studies, forensic analyses, and authentication of medicinal plants. The roots of Astragalus membranaceus and A. membranaceus var. mongholica are commonly used as Radix Astragali in several Asian countries, including China, Japan, and Korea. However, in addition to the two species recorded in the Chinese Pharmacopoeia, there are twenty-three species from different genera including Astragalus, Oxytropis, Hedysarum, and Glycyrrhiza, which have been used as adulterants not only in trading markets but also by the herbal medicine industry. Therefore, a simple, reliable, and accurate classification method is important for distinguishing authentic Radix Astragali from its adulterants. In this study, we acquired data for 37 samples from four related genera within the family Fabaceae. Then we compared four candidate DNA barcoding markers using ITS, matK, rbcL, and coxI sequences from nuclear, chloroplast, and mitochondrial genomes, all commonly used for plants to identify genetic variations among genera, intraspecies, and interspecies. We observed higher divergences among genera and interspecies for ITS, which have the average Kimura 2-parameter distances of 4.5% and 14.1%, respectively, whereas matK was found to have sufficient divergence at the intraspecific level. Moreover, two indels detected in the matK sequence are useful for PCR studies in distinguishing Radix Astragali from its adulterants. This study suggests that the combined barcoding regions of ITS and matK are superior barcodes for Radix Astragali and further studies should focus on evaluating the applicability and accuracy of such combined markers for a wide range of traditional Chinese herbs.

DNA Barcode ITS Effectively Distinguishes the Medicinal Plant Boerhavia diffusa from Its Adulterants

Boerhavia diffusa （B. diffusa）, also known as Punarnava, is an indigenous plant in India and an important component in traditional Indian medicine. The accurate identification and collection of this medicinal herb is vital to enhance the drug＇s efficacy and biosafety. In this study, a DNA barcoding technique has been applied to identify and distinguish B. diffusa from its closely-related species. The phylogenetic analysis was carried out for the four species of Boerhavia using barcode candidates including nuclear ribosomal DNA regions ITS, ITS1, ITS2 and the chloroplast plastid gene psbA-trnH. Sequence alignment revealed 26% polymorphic sites in ITS, 30% in ITS1, 16% in ITS2 and 6% in psbA-trnH, respectively. Additionally, a phylogenetic tree was constructed for 15 species using ITS sequences which clearly distinguished B. diffusa from the other species. The ITS1 demonstrates a higher transition/transversion ratio, percentage of variation and pairwise distance which differentiate B. diffusa from other species of Boerhavia. Our study revealed that 1TS and ITS1 could be used as potential candidate regions for identifying B. diffusa and for authenticating its herbal 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.

Study on Authentication technology of Notoginseng

Notoginseng has the effect of dispersing blood stasis, relieving swelling and pain. It is mainly used for cardiovascular and cerebrovascular diseases. At present, because of the wide application of Notoginseng, the market demands a great deal, leading to the emergence of a lot of fake products in the market as authentic Notoginseng, or adulterated in the powder of Notoginseng, which is a great potential harm to consumers. Therefore, the identification of Notoginseng becomes particularly important. Nowadays, scholars have developed a variety of methods for the identification of Notoginseng, including character identification, microscopic identification, physical and chemical identification, protein marker identification and molecular identification. This paper summarizes the research progress of the identification technology of Notoginseng and provides theoretical basis and method basis for the supervision of market of Notoginseng.

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.

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.

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 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.

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.

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.

Validation of Two Real-Time PCR Approaches for the Relative Quantitation of Pork and Horse DNA in Food Samples

Two real-time PCR methods for the relative quantitation of DNA from meat species in food samples are described: these methods are applicable for horse in processed beef meat products, and pork in raw/processed beef meat products. Test samples were prepared using raw meat admixtures or processed horse/pork in beef food products made to an industry-standard recipe. The methods were subjected to single laboratory method validation, evaluating the performance characteristics of specificity, PCR efficiency and r-squared (r2), Limit of Detection (LOD), Limit of Quantitation (LOQ), and precision and trueness. A limited UK-based inter-laboratory trial of the two methods was completed involving four participating laboratories. Full statistical analysis of the data qualified the applicability of the methods for accurate and sensitive trace-level analysis. The methods were deemed fit for purpose for reproducibly distinguishing between adventitious contamination at 0.1% (w/w), the level for further enforcement action at 1% (w/w), and a level representative of deliberate economically motivated adulteration (10% (w/w)). The data provided evidence that the precision of the two methods was applicable for qualitative and quantitative detection at topically important levels of adulteration. This work has added significant value to the current state of the art in quantitative determination of topical meat species adulteration, allowing analysts to distinguish between adventitious contamination and deliberate adulteration. The resulting methods described in this paper can easily be deployed and used by analytical laboratories for controls and due-diligence testing based on standard laboratory equipment.

Visible and Near-Infrared Spectroscopic Discriminant Analysis Applied to Identification of Soy Sauce Adulteration

The identification of soy sauce adulteration can avoid fraud, and protect the rights and interests of producers and consumers. Based on two measurement models (1 mm, 10 mm), the visible and near-infrared (Vis-NIR) spectroscopy combined with standard normal variate-partial least squares-discriminant analysis (SNV-PLS-DA) was used to establish the discriminant analysis models for adulterated and brewed soy sauces. Chubang soy sauce was selected as an identification brand (negative, 70). The adulteration samples (positive, 72) were prepared by mixing Chubang soy sauce and blended soy sauce with different adulteration rates. Among them, the “blended soy sauce” sample was concocted of salt water (NaCl), monosodium glutamate (C5H10NNaO5) and caramel color (C6H8O3). The rigorous calibration-prediction-validation sample design was adopted. For the case of 1 mm, five waveband models (visible, short-NIR, long-NIR, whole NIR and whole scanning regions) were established respectively;in the case of 10 mm, three waveband models (visible, short-NIR and visible-short-NIR regions) for unsaturated absorption were also established respectively. In independent validation, the models of all wavebands in the cases of 1 mm and 10 mm have achieved good discrimination effects. For the case of 1 mm, the visible model achieved the optimal validation effect, the validation recognition-accuracy rate (RARV) was 99.6%;while in the case of 10 mm, both the visible and visible-short-NIR models achieved the optimal validation effect (RARV = 100%). The detection method does not require reagents and is fast and simple, which is easy to promote the application. The results can provide valuable reference for designing small dedicated spectrometers with different measurement modals and different spectral regions.