Risk Of Adulteration Research Articles

Impaction-Based Exhaled Breath Sampling for Substance Monitoring: A Prospective Pilot Study (DrugXhale).

For decades, intake monitoring of drugs using urine as the matrix of choice is the gold standard in drug treatment centers. A properly conducted urine drug test can identify recent use of prescribed, non-prescribed and illicit drugs. However, issues like adulteration, substitution and privacy issues have driven the search for alternative matrices. This prospective pilot study evaluates the use of an impaction-based breath sampling device, Breath Explor®, as an alternative to traditional urine-based drug monitoring. Breath samples were analyzed using a validated 32-component liquid chromatography-tandem mass spectrometry (LC-MSMS) method. Recovery data represent the efficiency of extracting the analytes from the breath devices. Both automated and manual processing of the Breath Explor® devices showed mean recovery rates ranging from 39.5% to 55.4% for the 32 analytes. Despite the small number of subjects, breath analysis proved to be a convenient and easy-to-use methodology. An overall kappa-values of 0.5 indicated a moderate level of agreement with urine analysis, underscoring its potential as a complementary diagnostic tool. All participants tested positive in their breath sample for methadone (70% methadone and 100% EDDP), while a significant portion (90%) tested positive for 6-monoacetylmorphine (6-MAM). This innovative approach offers several advantages, including non-invasiveness, reduced risk of adulteration, and the ability to perfom repeated automated sampling and confirmation testing. These findings suggest that breath-based substance monitoring could complement or even replace traditional urine-based methods in clinical practice.

Use of a novel combination of multiplex PCR and DNA barcoding in assessing authenticity of ginseng products

Ginseng (Panax sp.) is a medicinal plant used for its purported health benefits, primarily in East Asian countries. The COVID-19 pandemic led to a significant increase in sales of dietary supplements, including ginseng supplements, to purportedly support immune health and provide other health benefits. However, this heightened demand has subsequently increased the risk of adulteration in these dietary supplements. This study aimed to determine the efficacy of a novel combination of DNA barcoding and multiplex PCR to identify species in ginseng supplements. A total of 50 commercial ginseng supplements containing Panax ginseng, Panax quinquefolius, or Panax notoginseng were obtained from an online vendor. Two composite samples per product were subjected to DNA extraction followed by multiplex PCR and DNA barcoding with three genetic targets (i.e., rbcL, matK, and ITS2). Of the three DNA barcoding markers, ITS2 had the highest amplification success at 74%, and matK had the highest sequencing success at 60%. DNA barcoding alone identified species in 68% of products, while multiplex PCR had an overall 60% identification rate. The combination of both methods resulted in a 72% species identification rate. In addition to improving the overall identification rate, the combination of both methods allowed for greater species resolution and the detection of undeclared plant species. Future research should explore combining DNA-based methods with chemical-based approaches to improve ginseng detection capabilities and enable the quantification of undeclared adulterants.

Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry Combined with Chemometrics for Protein Profiling and Classification of Boiled and Extruded Quinoa from Conventional and Organic Crops

Quinoa is an Andean crop that stands out as a high-quality protein-rich and gluten-free food. However, its increasing popularity exposes quinoa products to the potential risk of adulteration with cheaper cereals. Consequently, there is a need for novel methodologies to accurately characterize the composition of quinoa, which is influenced not only by the variety type but also by the farming and processing conditions. In this study, we present a rapid and straightforward method based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to generate global fingerprints of quinoa proteins from white quinoa varieties, which were cultivated under conventional and organic farming and processed through boiling and extrusion. The mass spectra of the different protein extracts were processed using the MALDIquant software (version 1.19.3), detecting 49 proteins (with 31 tentatively identified). Intensity values from these proteins were then considered protein fingerprints for multivariate data analysis. Our results revealed reliable partial least squares-discriminant analysis (PLS-DA) classification models for distinguishing between farming and processing conditions, and the detected proteins that were critical for differentiation. They confirm the effectiveness of tracing the agricultural origins and technological treatments of quinoa grains through protein fingerprinting by MALDI-TOF-MS and chemometrics. This untargeted approach offers promising applications in food control and the food-processing industry.

Authenticity and adulteration identification of Bubali Cornu based on DNA fingerprints

Whether adulteration exists is a difficult problem in the identification of traditional Chinese medicine(TCM). Bubali Cornu is mainly available in the medicinal material market in the form of buffalo horn silk or buffalo horn powder but lacks obvious identification characteristics, so there is a risk of adulteration. However, the method of identification of adulteration in Bubali Cornu is lacking at present. In order to ensure authenticity and identify adulteration of TCM Bubali Cornu, control the quality of TCM Bubali Cornu, and ensure the authenticity of clinical use, the DNA fingerprints of 43 batches of samples from pharmaceutical companies and medicinal material markets were identified, and the amplification primers of fluorescent DNA fingerprints of Bubali Cornu and Bovis Grunniens Cornu were screened. The DNA fingerprints of Bubali Cornu were obtained by fluorescent capillary typing. The identification effect of fluorescent capillary typing on different adulteration ratios was also tested. Two pairs of fluorescent STR typing primers, namely 16Sa and CRc, which can distinguish Bubali Cornu and Bovis Grunniens Cornu, were screened out, and a DNA fingerprint identification method was established. The 16Sa migration peaks of Bovis Grunniens Cornu and Bubali Cornu were 223.4-223.9 bp and 225.5-226.1 bp. The CRc migration peaks of Bovis Grunniens Cornu and Bubali Cornu were 518.8-524.8 bp and 535.9-542.5 bp. The peak height of the migration peak could be used for preliminary quantification of the adulterants with an adulteration ratio below 50%, and the quantitative results were similar to the adulteration ratio. In this study, a simple and quick universal DNA fingerprint method was established for the identification of Bubali Cornu and its adulterants, which could realize the identification of TCM Bubali Cornu and the semi-quantitative identification of the adulterants.

Synthesis of bioengineered heparin chemically and biologically similar to porcine-derived products and convertible to low MW heparin

Heparins have been invaluable therapeutic anticoagulant polysaccharides for over a century, whether used as unfractionated heparin or as low molecular weight heparin (LMWH) derivatives. However, heparin production by extraction from animal tissues presents multiple challenges, including the risk of adulteration, contamination, prion and viral impurities, limited supply, insecure supply chain, and significant batch-to-batch variability. The use of animal-derived heparin also raises ethical and religious concerns, as well as carries the risk of transmitting zoonotic diseases. Chemoenzymatic synthesis of animal-free heparin products would offer several advantages, including reliable and scalable production processes, improved purity and consistency, and the ability to produce heparin polysaccharides with molecular weight, structural, and functional properties equivalent to those of the United States Pharmacopeia (USP) heparin, currently only sourced from porcine intestinal mucosa. We report a scalable process for the production of bioengineered heparin that is biologically and compositionally similar to USP heparin. This process relies on enzymes from the heparin biosynthetic pathway, immobilized on an inert support and requires a tailored N-sulfoheparosan with N-sulfo levels similar to those of porcine heparins. We also report the conversion of our bioengineered heparin into a LMWH that is biologically and compositionally similar to USP enoxaparin. Ultimately, we demonstrate major advances to a process to provide a potential clinical and sustainable alternative to porcine-derived heparin products.

DNA barcoding of herbal supplements on the US commercial market associated with the purported treatment of COVID-19.

The COVID-19 pandemic was associated with an increased global use of traditional medicines, including Ayurvedic herbal preparations. Due to their growing demand, their processed nature, and the complexity of the global supply chain, there is an increased risk of adulteration in these products. The objective of this study was to assess the use of DNA barcoding for species identification in herbal supplements on the US market associated with the Ayurvedic treatment of respiratory symptoms. A total of 54 commercial products containing Ayurvedic herbs were tested with four DNA barcoding regions (i.e., rbcL, matK, ITS2, and mini-ITS2) using two composite samples per product. Nine categories of herbs were targeted: amla, ashwagandha, cinnamon, ginger, guduchi, tribulus, tulsi, turmeric, and vacha. At least one species was identified in 64.8% of products and the expected species was detected in 38.9% of products. Undeclared plant species, including other Ayurvedic herbs, rice, and pepper, were detected in 19 products, and fungal species were identified in 12 products. The presence of undeclared plant species may be a result of intentional substitution or contamination during harvest or processing, while fungal DNA was likely associated with the plant material or the growing environment. The greatest sequencing success (42.6-46.3%) was obtained with the matK and rbcL primers. The results of this study indicate that a combination of genetic loci should be used for DNA barcoding of herbal supplements. Due to the limitations of DNA barcoding in identification of these products, future research should incorporate chemical characterization techniques.

Bacteriological evaluation and advanced SYBR-green multiplex real-time PCR assay for detection of minced meat adulteration.

Minced meat is a valuable source of nutrients, but it is vulnerable to contamination by microorganisms commonly present in the environment. In addition, there is a risk of adulteration with cheaper meat sources, which can be harmful to consumers. It is crucial to identify meat adulteration with distinct microbiological analysis for legal, economic, religious, and public health purposes. A total of 100 minced meat samples were collected from several markets in Sharkia Governorate, Egypt. These samples were then subjected to bacteriological testing and an advanced multiplex PCR method. This method enables the detection of bovine, equine, porcine, and dog species in meat samples with just one step. The adulterated samples had a higher total bacterial count and pH values compared to pure bovine meat. These differences in bacterial count and pH values were statistically significant, with p-values of 0.843 (log10) and 0.233, respectively. The frequency of Escherichia coli occurrence was 13%, and the O111 serotype was predominant in the adulterated samples. Listeria monocytogenes and Staphylococcus aureus were isolated with prevalence rates of 3% and 29%, respectively. Besides, the SYBR-green multiplex real-time PCR assay used in this study detected adulteration with dog, equine, and porcine meats in the examined samples at rates of 9%, 5%, and 4%, respectively. This method provides a sensitive and specific approach to detect issues related to well-being and safety.

Estimating the extent of adulteration of the popular herbs black cohosh, echinacea, elder berry, ginkgo, and turmeric - its challenges and limitations.

Covering: up to July 2023Botanical natural medicinal products and dietary supplements are utilized globally for their positive impacts on health and wellness. However, the effectiveness and safety of botanical products can be compromised by unintentional or intentional adulteration. The presence of adulterated botanical ingredients in the global market has been documented in the published literature but a key question, namely what the extent of adulteration is, remains to be answered. This review aims to estimate the prevalence of adulteration in preparations made from black cohosh rhizome, echinacea root or herb, elder berry, ginkgo leaf, and turmeric root/rhizome. According to the information provided in the 78 publications retrieved for this paper, 818 of 2995 samples were reported to be adulterated and/or mislabeled. Ginkgo leaf samples (n = 533) had the highest adulteration rate with 56.7%, followed by black cohosh rhizome (n = 322) samples with 42.2%, echinacea root/herb (n = 200) with 28.5%, elder berry (n = 695) with 17.1%, and turmeric root/rhizome (n = 1247) with 16.5%. Products sold as licensed or registered herbal medicines were found to have a lower risk of adulteration compared to products sold as dietary/food supplements. The data show that the adulteration rate substantially differs from one ingredient to the other. Due to the significant limitations of the available data upon which the estimated extent of adulteration is based, and the rapidly changing botanical dietary supplement market, conclusions from the five herbs examined in this publication cannot be applied to other botanicals traded in the global market. However, the data clearly show that a substantial portion of the botanical dietary supplements do not contain what is claimed on their labels.

Two flavors in adulterated sesame oil: discovery, confirmation, and content regularity study.

Exploring and accurately detecting new adulteration markers in sesame oil is an important measure for sesame oil adulteration monitoring. In this study, two endogenous flavors sulfurol and γ-nonalactone which can be used as potential adulteration markers were first discovered in sesame oil and accurately quantified. First, the two endogenous flavors were discovered using gas chromatography-mass spectrometry (GC-MS), and their structures were confirmed by comparing the mass spectrograms with the NIST spectral library. Then the liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using direct methanol extraction pretreatment and vanillin-D3 as an internal standard was developed for rapid quantitation and application. The method was successfully validated with recoveries ranging from 88.5% to 102.2% and relative standard deviations between 2.6% and 10.5% (n = 6). The combined method of GC-MS and LC-MS/MS was indicated to be efficient and highly sensitive for detection of sulfurol and γ-nonalactone in edible oil. Subsequently, 31 sesame oils from the market were detected, revealing that 31 samples contained the identified flavors within a relatively consistent range. However, the concentration of these flavor substances in one sample was abnormally high, indicating that there was a potential risk of adulteration. Therefore, the developed method shows good potential for quality evaluation and adulteration screening of sesame oil.

Rapid Detection of Adulteration in Boswellia Extracts with Citric Acid by UPLC–HRMS and 1H NMR

Boswellia serrata ole-gum-resin extracts (BSEs) are commonly used as food supplements, especially in osteoarthritis management. The quality standard is established by determining 11-keto-β-boswellic acid (KBA) and acetyl-11-keto-boswellic acid (AKBA) content using high-performance liquid chromatography (HPLC) or assessing the total boswellic acid (TBA) content by titrimetry. The limited geographical distribution of Boswellia species and increasing industrial demand could increase the risk of adulteration in Boswellia-containing products. In this study, 14 BSEs from commercial sources, used in food supplements, were analyzed in comparison with a USP Reference Standard extract. The KBA and AKBA content was determined by HPLC, whereas the TBA content was determined by titration. Targeted UHPLC-high-resolution mass spectrometry (HRMS) was applied to identify the carboxylic acid content in the samples. The 1H NMR spectra of extracts were also analyzed. Only two products met the criteria for KBA and AKBA content. Although, the TBA content complied with the expected amount, 10 extracts contained citric acid levels of 6–11% even though citric acid is not a cha­racteristic component of BSEs. Our results suggest undeclared addition of citric acid to comply with declared contents of TBA when using titration methods. Incorporation of citric acid to industrial samples – in order to alter the outcomes of the titration analysis – was demonstrated for the first time.

Drug Screening of 49 Compounds by Liquid Chromatography-Tandem Mass Spectrometry in Saliva.

In recent years, the use of oral fluid as a testing matrix for drug analysis has become increasingly popular due to its benefits, such as a possibility for direct observation of a sample collection and, thus, a lower risk of adulteration. To address this trend, we have developed a quantitative LC-MS/MS method that can simultaneously analyze 49 commonly prescribed compounds in oral fluid. The assay is performed on an AB Sciex 4500 electrospray ionization mass spectrometer in multiple reaction monitoring mode.

What are the challenges for implementing an "organic label" to camel milk?

Increasing demand for camel's milk worldwide occurred in the context of the development of the organic sector in agriculture. The implementation of an organic label for camel milk has never been established. However, the creation of such a label faces to important challenges that are investigated in the present paper. Indeed, although camel milk conveys the image of a "natural product" issued from remote places, the risk of being produced in contaminated areas (mining activities, oil extraction) cannot be neglected for grazing animals. Moreover, the management of veterinary drugs for prevention or curative treatment can lead to the presence of residues in milk, especially in camel species with different pharmacokinetics, although similar instructions than for cow milk are used. Moreover, the lack of international standards regarding both composition and hygienic rules, the risks of adulteration, and the necessity to use specific indicators or analytical procedures adapted to the behavior of camel milk, have to be taken in account in the establishment of the specifications for the camel milk producers through the world.

DNA Barcoding Based Identification of Rosa x Damascene and Prunus dulcis Herbs Using ITS2 Barcoding Gene Amplification.

The considerable risk of adulteration in the herbs has raised commercial interest in the identification of medicinal herbs globally. DNA barcoding is the primary techniques for identifying the herbs at genetic level. This technique's key benefit is that it can identify the material's purity. This study focuses on the accurate identification of species utilizing Polymerase chain reaction-based nuclear universal internal transcribed spacer region (a barcode region) amplification and sequencing in 2 medically significant plants (Rosa damascene and Prunus dulcis) procured from the local herbal market of Karachi in the year 2022. Results suggest that the ribosomal nuclear ITS2 region of the selected plant species shows the 100 % identity with the reference genome, therefore it has shown a good rate of identification at the species level. The findings of this concludes that ITS2 is the novel standard barcode that involve in species identification, genomic conservation, and secure utilization of the medically significant plant species.

Rapid colorimetric and fluorescence identification of Pinelliae Rhizoma and adulterate Rhizoma Typhonii Flagelliformis using direct-LAMP assay

Pinelliae Rhizoma, renowned for its medicinal benefits against cough, insomnia and inflammation, is frequently utilized in botanical dietary supplements and herbal products. However, due to high demand, there is a risk of adulteration. This study introduced Direct-Loop-Mediated Isothermal Amplification (Direct-LAMP), a rapid colorimetric technique, for distinguishing Pinelliae Rhizoma from its primary adulterant, Rhizoma Typhonii Flagelliformis. This method comprises three parts: DNA extraction-free direct lysis, isothermal amplification and prompt color development. The direct lysis process allows for release of genomic DNA within 2 min. The isothermal amplification reaction exhibits excellent analytical performance, detecting Pinelliae Rhizoma at levels as low as 0.1 pg and Rhizoma Typhonii Flagelliformis at 0.08 ng, while also demonstrating good linearity across a wide range, making it suitable for quantitative analysis. Two different color reagents are employed for these two different plant materials, resulting in dual-color changes for captivating visual detection. The developed direct-LAMP was also successfully validated across 48 actual samples.

Label-free detection of porcine gelatin: A reliable immunosensor based on multi-walled carbon nanotubes and gold nano-urchins

Gelatin derived from porcine sources raises significant ethical and religious concerns worldwide, particularly due to its increasing use in food-based products without adequate monitoring and control. Once gelatin is incorporated into processed food items, determining its source animal becomes challenging, which consequently increases the risks of adulteration and mislabeling. In this study, we propose the utilization of a novel nanocomposite comprising multi-walled carbon nanotubes (MWCNTs), chitosan (CS), and gold nano-urchins (AuNUs) for constructing a label-free electrochemical immunosensor. The purpose of this immunosensor is to effectively detect Porcine skin gelatin (PSG) in food samples using differential pulse voltammetry (DPV) with a 5 mM [K3Fe(CN6)] redox probe. Under optimized conditions, the fabricated biosensor demonstrates superior sensitivity to PSG, exhibiting a linear detection range of 50 pg/mL to 1000 a pg/mL and a limit of detection (LOD) of 23.079 pg/mL. Moreover, the constructed biosensor exhibits excellent reproducibility, acceptable selectivity, and interference resistance. Notably, it achieves outstanding recovery rates when applied to real food samples spiked with PSG, thus indicating its potential for practical implementation in food safety monitoring.

Opioid-related harms and care impacts of conventional and AI-based prescription management strategies: insights from leveraging agent-based modeling and machine learning.

Like its counterpart to the south, Canada ranks among the top five countries with the highest rates of opioid prescriptions. With many suffering from opioid use disorder first having encountered opioids via prescription routes, practitioners and health systems have an enduring need to identify and effectively respond to the problematic use of opioid prescription. There are strong challenges to successfully addressing this need: importantly, the patterns of prescription fulfillment that signal opioid abuse can be subtle and difficult to recognize, and overzealous enforcement can deprive those with legitimate pain management needs the appropriate care. Moreover, injudicious responses risk shifting those suffering from early-stage abuse of prescribed opioids to illicitly sourced street alternatives, whose varying dosage, availability, and the risk of adulteration can pose grave health risks. This study employs a dynamic modeling and simulation to evaluate the effectiveness of prescription regimes employing machine learning monitoring programs to identify the patients who are at risk of opioid abuse while being treated with prescribed opioids. To this end, an agent-based model was developed and implemented to examine the effect of reduced prescribing and prescription drug monitoring programs on overdose and escalation to street opioids among patients, and on the legitimacy of fulfillments of opioid prescriptions over a 5-year time horizon. A study released by the Canadian Institute for Health Information was used to estimate the parameter values and assist in the validation of the existing agent-based model. The model estimates that lowering the prescription doses exerted the most favorable impact on the outcomes of interest over 5 years with a minimum burden on patients with a legitimate need for pharmaceutical opioids. The accurate conclusion about the impact of public health interventions requires a comprehensive set of outcomes to test their multi-dimensional effects, as utilized in this research. Finally, combining machine learning and agent-based modeling can provide significant advantages, particularly when using the latter to gain insights into the long-term effects and dynamic circumstances of the former.

Awareness that cocaine can contain fentanyl among nightclub and festival attendees in New York City, 2018-2022.

People who attend nightclubs and dance festivals-particularly those that feature electronic dance music (EDM), are at high risk for use of cocaine and other party drugs. Given that cocaine is now sometimes adulterated with fentanyl, this study examines trends in people's knowledge about such risk of adulteration to inform prevention and harm reduction efforts. Adults were surveyed entering randomly selected EDM events (including dance festivals) in New York City in 2018, 2019, 2021, and 2022 (n=2107). They were asked whether they agree that some dealers sell cocaine containing fentanyl, and trends in agreement were estimated. Prevalence of agreement that cocaine can be adulterated increased from 42.1% (95% CI: 36.8-47.6) in 2018 to 65.6% (95% CI: 54.1-75.4) in 2022-a 55.8% increase (p<.001). Between 2018 and 2022, particular increases occurred among those not reporting past-year cocaine use (by 61.6%, p<.001), among White individuals (by 68.1%, p<.001), those with some college (by 68.5%, p=.021), and those age ≥26 (by 83.8%, p=.001). Awareness that cocaine can contain fentanyl is increasing in this high-risk population. Continued education is needed for high-risk populations regarding both opioid overdose response and test strips that can test drugs for the presence of fentanyl.

Quality control of marketed herbal products of Asparagus racemosus Willd. through high performance thin layer chromatography (HPTLC) analysis

Asparagus racemosus Willd. is a valuable medicinal plant which is used all over the world. There are several marketed products of A. racemosus. The high demand for this herb has increased the risk of adulteration in its commercial products. The adulterated herbal products might pose serious ill effects on health. Therefore, it is necessary to check the quality of marketed products in terms of the presence of their major bioactive compounds. The present study aimed to carry out the qualitative and quantitative analysis of Shatavarin IV in marketed products of A. racemosus through a validated high performance thin layer chromatography (HPTLC) method. Ten marketed products were analysed and all of them had shown the presence of Shatavarin IV which was quantified. The identification and quantification were done by taking a standard Shatavarin IV as reference. The Shatavarin IV was detected at Rf 0.4±0.05 and showed maximum absorption at 425 nm. The Shatavarin IV was quantified using a 6-point calibration curve having a standard deviation of 3.89 % with an R2 value of 0.9968. The amount of Shatavarin IV varied between 1.47±0.25 to 2.69±0.51 mg/g on a dry weight basis which is a normal range in the raw plant materials. Thus, the present findings would be a simple, reliable and cost-effective method for the quality determination of herbal products of A. racemosus. The developed HPTLC chromatograms would serve as a reference for the quality assessment of commercial products of A. racemosus in future.

Development of a Smart Sensor Array for Adulteration Detection in Black Pepper Seeds using Machine Learning

Black pepper is an expensive commodity with a high risk of adulteration. Ground papaya seed is the main adulterant in pepper because it cannot be discriminated visually. There are few destructive methods. Since pepper is costlier, non-destructive method of adulteration is must but it is challenging one. The existing non-destructive method uses costlier equipment, bulky, involve laboratory-based testing, time consuming in the process. To overcome the above issues, this article presents the development of Non-destructive E- nose gas sensor for pepper adulteration detection. This system determines the VOC in a controlled environment. The proposed system utilizes MQ2 and MQ3 gas sensor arrays to identify Volatile Organic Compounds present in pepper seeds to discriminate adulterant and non-adulterant sample. The sensor data are utilized to perform the qualitative analysis to determine the adulteration using a support vector machine learning algorithm. The proposed sensor system with Support Vector Machine learning algorithm outperforms in comparison with existing methods with 100% classification accuracy. Conclusion: The developed gas sensor system is connected to the internet via the IoT application model to show results on the web pages and enables access by the authenticated user from anywhere. Client server model with MQTT protocol is used for developing IoT application.

A Heptaplex PCR Assay for Molecular Traceability of Species Origin With High Efficiency and Practicality in Both Raw and Heat Processing Meat Materials.

Frequent meat frauds have become a global issue because adulteration risks the food safety, breaches market rules, and even threatens public health. Multiplex PCR is considered to be a simple, fast, and inexpensive technique that can be applied for the identification of meat products in food industries. However, relatively less is known about a multiplex PCR method authenticating seven animal species simultaneously in one reaction due to technological challenge. Through screening new species-specific primers and optimizing PCR system, a heptaplex PCR method was established, which could simultaneously detect seven meat ingredients of camel (128 bp), pigeon (157 bp), chicken (220 bp), duck (272 bp), horse (314 bp), beef (434 bp), and pork (502 bp) in a single-tube reaction. DNA sequencing solidly validated that each set of primers specifically amplified target species from total DNA mixtures of seven meat species. The developed multiplex assay was stable and sensitive enough to detect 0.01–0.025 ng DNA from various meat treatments including raw, boiled, and autoclaved meat samples or target meat content of 0.1% total meat weight, suggesting the suitability of the heptaplex PCR technique for tracing target meats with high accuracy and precision. Most importantly, a market survey validated the availability of this multiplex PCR technique in real-world meat products with a good application foreground.