Saffron Authentication Research Articles

Authentication of saffron spice accessions from its common substitutes via a multiplex approach of UV/VIS fingerprints and UPLC/MS using molecular networking and chemometrics

Saffron is a spice revered for its unique flavor and health attributes often subjected to fraudulence. In this study, molecular networking as a visualization tool for UPLC/MS dataset of saffron and its common substitutes i.e. safflower and calendula (n = 21) was employed for determining genuineness of saffron and detecting its common substitutes i.e. safflower and calendula. Saffron was abundant in flavonol-O-glycosides and crocetin esters versus richness of flavanones/chalcones glycosides in safflower and cinnamates/terpenes in calendula. OPLS-DA identified differences in UPLC/MS profiles of different saffron accessions where oxo-hydroxy-undecenoic acid-O-hexoside was posed as saffron authentication marker and aided in discrimination between Spanish saffron of high quality from its inferior grade i.e. Iranian saffron along with crocetin di-O-gentiobiosyl ester and kaempferol-O-sophoroside. Kaempferol-O-neohesperidoside and N,N,N,-p-coumaroyl spermidine were characteristic safflower metabolites, whereas, calendulaglycoside C and di-O-caffeoyl quinic acid were unique to calendula. UV/VIS fingerprint spectral regions of picrocrocin (230–260 nm) and crocin derivatives (400–470 nm) were posed as being discriminatory of saffron authenticity and suggestive it can replace UPLC/MS in saffrom quality determination.

Optimization of an Untargeted DART-HRMS Method Envisaging Identification of Potential Markers for Saffron Authenticity Assessment.

Saffron is one of the most expensive agricultural products in the world and as such, the most commonly adulterated spice, with undeclared plant-based surrogates or synthetic components simulating color and morphology. Currently, saffron quality is certificated in the international trade market according to specific ISO guidelines, which test aroma, flavor, and color strength. However, it has been demonstrated that specific adulterants such as safflower, marigold, or turmeric up to 20% (w/w) cannot be detected under the prescribed approach; therefore, there is still a need for advanced and sensitive screening methods to cope with this open issue. The current investigation aims to develop a rapid and sensitive untargeted method based on an ambient mass spectrometry ionization source (DART) and an Orbitrap™high-resolution mass analyzer to discriminate pure and adulterated saffron samples with either safflower or turmeric. The metabolic profiles of pure and adulterated model samples prepared at different inclusion levels were acquired. Unsupervised multivariate analysis was carried out based on hierarchical cluster analysis and principal component analysis as first confirmation of the discriminating potential of the metabolic profile acquired under optimized DART-HRMS conditions. In addition, a preliminary selection of potential markers for saffron authenticity was accomplished, identifying compounds able to discriminate the type of adulteration down to a concentration level of 5%.

External parameter orthogonalization-support vector machine for processing of attenuated total reflectance-mid-infrared spectra: A solution for saffron authenticity problem

In the present work, a new approach based on external parameter orthogonalization combined with support vector machine (EPO-SVM) is proposed for processing of attenuated total reflectance-Fourier transform mid-infrared (ATR-FT-MIR) spectra with the goal of solving authentication problem in saffron, the most expensive spice in the world. First, one-hundred authentic saffron samples are clustered by principal component analysis (PCA) with EPO as the best preprocessing strategy. Then, EPO-SVM is used for the detection of four commonly used plant-derived adulterants (i.e. safflower, calendula, rubia, and style) in binary mixtures (saffron and each of plant adulterants) and its performance is compared with other common classification methods. The obtained results showed that the EPO-SVM approach has a much better classification accuracy (>95%) than other methods (accuracy<89.2%). Finally, two different sample sets including mixture of saffron and four plant adulterants and commercial saffron samples are used for validation of the developed EPO-SVM model. In this regard, classification figures of merit in terms of sensitivity, specificity and accuracy were respectively 96.6%, 97.1%, and 96.8% which showed good classification performance. It is concluded that the proposed EPO-PCA and EPO-SVM approaches can be considered as reliable tools for authentication and adulteration detection in saffron samples.

Comparison of near-infrared (NIR) and mid-infrared (MIR) spectroscopy based on chemometrics for saffron authentication and adulteration detection

In this work, the potential of near-infrared (NIR) and mid-infrared (MIR) spectroscopy along with chemometrics was investigated for authentication and adulteration detection of Iranian saffron samples. First, authentication of one-hundred saffron samples was examined by principal component analysis (PCA). The results showed the NIR spectroscopy can better predict the origin of samples than the MIR. Next, partial least squares-discriminant analysis (PLS-DA) was developed to detect four common plant-derived adulterants (i.e., saffron style, calendula, safflower, and rubia). In all cases, PLS-DA classification figures of merit in terms of sensitivity, specificity, error rate and accuracy were satisfactory for both NIR and MIR datasets. The built models were then successfully validated using test set and also commercial samples. Finally, partial least squares regression (PLSR) was used to estimate the amount of adulteration. In this case, only NIR showed a good performance with regression coefficients (R2) in range of 0.95–0.99.

Combining multivariate image analysis with high-performance thin-layer chromatography for development of a reliable tool for saffron authentication and adulteration detection.

In this work, high-performance thin-layer chromatography (HPTLC) coupled with multivariate image analysis (MIA) is proposed as a fast and reliable tool for authentication and adulteration detection of Iranian saffron samples based on their HPTLC fingerprints. At first, the secondary metabolites of saffron were extracted using ultrasonic-assisted solvent extraction (UASE) which was optimized using central composite design (CCD). Next, the RGB coordinates of HPTLC images were used for estimation of saffron origin based on principal component analysis (PCA). The PCA scores plot showed that saffron samples were clustered into two clear-cut groups which was 92% matched with the geographical origins of the samples. In the next step, common plant-derived adulterants of saffron including safflower, saffron style, calendula, and rubia were investigated with MIA analysis of HPTLC images using partial least squares-discriminant analysis (PLS-DA) at 5-35% (w/w) levels. The PLS-DA results showed proper classification of saffron and adulterants with sensitivity 99.14%, specificity 96.94%, error rate 1.96% and accuracy 98.04. Also, the effect of HPTLC injection volume on the performance of the proposed strategy was evaluated. The ability of the proposed method was then investigated by analyzing two additional sample sets including mixed samples of four plant-derived adulterants and adulterated commercial samples. Sensitivity and specificity of this model were 100% which confirmed its validity.

Identification of authenticity, quality and origin of saffron using hyperspectral imaging and multivariate spectral analysis

This paper is conducted to identify the authenticity, quality, and origin of saffron using hyperspectral imaging and multivariate spectral analysis. Reflectance spectra were extracted from hyperspectral images of saffron. Successive projections algorithm, genetic algorithm, uninformative variable elimination, and competitive adaptive reweighted sampling were used to select characteristic wavelengths. Back propagation neural network model was established based on the selected wavelengths. Results showed that the model combining competitive adaptive reweighted sampling with back propagation neural network achieved the best performance. Its prediction accuracy of the one-adulterated, three-domestic and two-imported saffron was 100, 95, 94, 100, 83, and 96%, respectively.

Authentication of PDO saffron of L'Aquila (Crocus sativus L.) by HPLC-DAD coupled with a discriminant multi-way approach

One hundred and forty-nine (149) Italian saffron samples produced in the years from 2013 to 2016 in distinct sites located in five different Italian regions, Abruzzo, Tuscany, Umbria, Campania and Sardinia, together with twenty-seven (27) commercial samples, have been analyzed by high-performance liquid chromatography with diode array detection (HPLC-DAD). Among the investigated samples, those produced in Abruzzo (L'Aquila area) present an even higher added-value, because, since 2005, saffron of L'Aquila has been granted of the protected designation of origin (PDO) mark. In the present study, two different analytical approaches aimed at distinguishing PDO saffron of L'Aquila from the other samples have been compared. The first strategy is a more traditional approach, where the chromatograms collected at specific wavelengths are classified by Partial Least Squares Discriminant Analysis (PLS-DA). The second strategy exploits the multi-way nature of data, avoiding discarding any source of information. Consequently, the entire spectro-chromatogram is handled by N-Partial Least Squares (N-PLS) and then classified by Linear Discriminant Analysis (LDA). Both approaches provided satisfactory predictions; the best results from the prediction point of view (estimated on an external set of samples) were achieved by the proposed multi-way methodology.

A rapid MALDI MS/MS based method for assessing saffron (Crocus sativus L.) adulteration

We report on a sensitive and fast quantitative MALDI-MS/MS method used to assess saffron authenticity by direct analysis through the determination of picrocrocin as the saffron authenticity marker, and using curcumin as the non-isotopic isobaric internal standard. The internal standard curcumin yielded good linearity (R2 = 0.994), and with confidence intervals at 95% for intercept. The detectable maximum adulteration percentage (99.0%) was estimated interpolating the limit of detection (LOD) for the isobaric internal standard in linear regression. The LOD was 47.63 ppm, and LOQ was 56.53 ppm. Good accuracy and precision were obtained for all concentrations. The capability of the MS approach to monitor analytes in a specific, selective fashion was used to obtain a semi-quantitative adulteration percentage and to establish the adulterant by additional experiments. The detection of gardecin and its derivatives in commercial samples indicated that Gardenia jasminoides Ellis was used as the adulterant.

Identification of phenolic markers for saffron authenticity and origin: An untargeted metabolomics approach.

Saffron is a high-quality and expensive spice being widely subjected to adulteration. An UHPLC-ESI/QTOF-MS metabolomic-based approach was therefore used to investigate the discrimination potential between adulterated (added with different percentage of other parts of the flower) and authentic saffron, as well as to trace its geographical origin. Both unsupervised (hierarchical clustering) and supervised OPLS-DA multivariate statistics allowed discriminating authentic saffron from styles added of other floral components, as well as PDO (Protected Designation of Origin) vs non PDO saffron samples according to their chemical fingerprints. The proposed markers were then validated through ROC curves. Anthocyanins and glycosidic flavonols were the best markers of the styles' adulteration. However, other flavonoids (mainly free flavonols and flavones), together with protocatechuic aldehyde and isomeric forms of hydroxybenzoic acid, were also validated as markers for the discrimination of PDO vs non PDO saffron samples. This work outlines the potential of untargeted metabolomics based on UHPLC-ESI/QTOF mass spectrometry for saffron authenticity and traceability.

A Quick and Efficient Non-Targeted Screening Test for Saffron Authentication: Application of Chemometrics to Gas-Chromatographic Data.

Saffron is one of the most adulterated food products all over the world because of its high market prize. Therefore, a non-targeted approach based on the combination of headspace flash gas-chromatography with flame ionization detection (HS-GC-FID) and chemometrics was tested and evaluated to check adulteration of this spice with two of the principal plant-derived adulterants: turmeric (Curcuma longa L.) and marigold (Calendula officinalis L.). Chemometric models were carried out through both linear discriminant analysis (LDA) and partial least squares discriminant analysis (PLS-DA) from the gas-chromatographic data. These models were also validated by cross validation (CV) and external validation, which were performed by testing both models on pure spices and artificial mixtures capable of simulating adulterations of saffron with the two adulterants examined. These models gave back satisfactory results. Indeed, both models showed functional internal and external prediction ability. The achieved results point out that the method based on a combination of chemometrics with gas-chromatography may provide a rapid and low-cost screening method for the authentication of saffron.

Comparison of IRMS, GC-MS and E-Nose data for the discrimination of saffron samples with different origin, process and age

In this study the use of conventional (Isotope-ratio mass spectrometry-IRMS and solid-phase microextraction gas chromatography mass spectrometric, SPME-GC-MS) and non-conventional analytical techniques (Electronic Nose) to characterize and discriminate origin, drying and age of 35 saffron samples was investigated.The IRMS technique by the analysis of the stable carbon and nitrogen isotopes has proved to be a reliable method in discriminating the geographical origin of saffron. Taking into account the chemical classes of the detected volatile compounds, the SPME-GC-MS was able to discriminate the different origin, drying and age. An E-Nose was used as alternative and rapid tool to characterize the complex aroma patterns and to exploit authenticity of saffron samples. Overall, the innovative AuNP-peptide based sensors array showed only a good discrimination of their origin.Results of this study could contribute to select and identify routine quality control methods for quality and authentication of saffron.

MARKET SAMPLE SURVEY OF CROCUS SATIVUS LINN. TO ASSESS THE GENUINITY FOR USING ANTIOXIDANT ACTIVITY, TOTAL PHENOLIC CONTENT AND HIGH-PRESSURE THIN LAYER CHROMATOGRAPHY USING DETECTION OF FLAVONOIDS

Objective: Herbalism is a traditional medicine or folk medicine practice based on the use of plants and plant extracts. Many of the drugs used in conventional medicine are dried from herbs. Despite the fluctuation in prices in international markets, saffron was still remained the most expensive spice. The main aim of this study is to examine the antioxidant activity, total phenolic content, high-pressure thin layer chromatography using flavanoid analysis and adulteration detection of saffron. Crocus sativus. Linn is a perennial stemless herb of the Iridaceae family. Saffron stigmas of sample1, sample2, sample3and sample4 are collected from different rates of the market sample from Thrissur district, sample5 collected from the Oushadhi premises, and it is collected from Himachal Pradesh.&#x0D; Methods: In this study detecting the antioxidant activity, total phenolic content, high-pressure thin layer chromatography using flavanoid analysis of different samples of saffron stigmas. The extracts were prepared by using ethanol as a solvent.&#x0D; Results: Safranal is present only in s5 sample. It is the main essential volatile oil responsible for the saffron characteristic such as odour. Phenolic content is varied in different market samples. The amount of phenolic compounds in the saffron extract was determined using the Folin-ciocalteau reagent. Total phenolic content is the help to detect the pure and fake saffron. The phenolic content is higher in S5. Sample S5 showed 0.737 mg/ml phenolic content. Lowest level of phenolic content in sample S3. Sample S3 showed 0.0887 mg/ml phenolic content. Sample S4 showed 0.564 mg/ml total phenolic content. Sample S1 showed 0.416 mg/ml total phenolic content and sample S2 showed 0.267 mg/ml phenolic content. Antioxidant activity is higher in sample s5. and it is different in different market samples. Sample 5 stigma posses higher antioxidant activity. Sample S5 showing 14.88% antioxidant activity in 100 mg/ml concentration, 7.26% in 80 mg/ml concentration, 2.23% in 60 mg/ml concentration, 2.21% in 40 mg/ml and 1.01% in 20 mg/ml concentration. Sample S3 showed the lower antioxidant activity in 0.1% in 60 mg/ml concentration and 0.1% in 80 mg/ml. Ascorbic acid standard showing 14.89% in 100 mg/ml concentration, 7.26% in 80 mg/ml concentration, 4.56% in 60 mg/ml concentration, and 3.1% in 40 mg/ml concentration, and 1% in 20 mg/ml concentration. Flavonoid content is different in different samples. It is present highly present in sample s1 and s5. sample s3 do not contain the Flavanoid. The quality of the samples depend on the price values.&#x0D; Conclusion: The authenticity of saffron is an extremely important matter for the industry and for the consumers in view of security and protection,quality assurance, active properties and last but not least, economic impact. Despite the fluctuation in prices in international markets, saffron was and still remains the most expensive spice. The genuine saffron samples possess higher price value. The fake saffron available in the market with lower price value. The quality of the saffron depends upon the price values. These observations would be of immense value in the botanical identification and standardization of the drug in crude form and would help to distinguish the drug from its other spices.

MARKET SAMPLE SURVEY OF CROCUS SATIVUS LINN. TO ASSESS THE GENUINITY FOR USING ANTIOXIDANT ACTIVITY, TOTAL PHENOLIC CONTENT AND HIGH-PRESSURE THIN LAYER CHROMATOGRAPHY USING DETECTION OF FLAVONOIDS

Objective: Herbalism is a traditional medicine or folk medicine practice based on the use of plants and plant extracts. Many of the drugs used in conventional medicine are dried from herbs. Despite the fluctuation in prices in international markets, saffron was still remained the most expensive spice. The main aim of this study is to examine the antioxidant activity, total phenolic content, high-pressure thin layer chromatography using flavanoid analysis and adulteration detection of saffron. Crocus sativus. Linn is a perennial stemless herb of the Iridaceae family. Saffron stigmas of sample1, sample2, sample3and sample4 are collected from different rates of the market sample from Thrissur district, sample5 collected from the Oushadhi premises, and it is collected from Himachal Pradesh.&#x0D; Methods: In this study detecting the antioxidant activity, total phenolic content, high-pressure thin layer chromatography using flavanoid analysis of different samples of saffron stigmas. The extracts were prepared by using ethanol as a solvent.&#x0D; Results: Safranal is present only in s5 sample. It is the main essential volatile oil responsible for the saffron characteristic such as odour. Phenolic content is varied in different market samples. The amount of phenolic compounds in the saffron extract was determined using the Folin-ciocalteau reagent. Total phenolic content is the help to detect the pure and fake saffron. The phenolic content is higher in S5. Sample S5 showed 0.737 mg/ml phenolic content. Lowest level of phenolic content in sample S3. Sample S3 showed 0.0887 mg/ml phenolic content. Sample S4 showed 0.564 mg/ml total phenolic content. Sample S1 showed 0.416 mg/ml total phenolic content and sample S2 showed 0.267 mg/ml phenolic content. Antioxidant activity is higher in sample s5. and it is different in different market samples. Sample 5 stigma posses higher antioxidant activity. Sample S5 showing 14.88% antioxidant activity in 100 mg/ml concentration, 7.26% in 80 mg/ml concentration, 2.23% in 60 mg/ml concentration, 2.21% in 40 mg/ml and 1.01% in 20 mg/ml concentration. Sample S3 showed the lower antioxidant activity in 0.1% in 60 mg/ml concentration and 0.1% in 80 mg/ml. Ascorbic acid standard showing 14.89% in 100 mg/ml concentration, 7.26% in 80 mg/ml concentration, 4.56% in 60 mg/ml concentration, and 3.1% in 40 mg/ml concentration, and 1% in 20 mg/ml concentration. Flavonoid content is different in different samples. It is present highly present in sample s1 and s5. sample s3 do not contain the Flavanoid. The quality of the samples depend on the price values.&#x0D; Conclusion: The authenticity of saffron is an extremely important matter for the industry and for the consumers in view of security and protection,quality assurance, active properties and last but not least, economic impact. Despite the fluctuation in prices in international markets, saffron was and still remains the most expensive spice. The genuine saffron samples possess higher price value. The fake saffron available in the market with lower price value. The quality of the saffron depends upon the price values. These observations would be of immense value in the botanical identification and standardization of the drug in crude form and would help to distinguish the drug from its other spices.

A rapid site-specific PCR based one-tube authentication method for saffron in crude drugs and processed herbal medicines.

Saffron is one of the world's most precious medicinal herbs and often found to be adulterated with other cheaper materials. The chemical compounds in Crocus sativus L. such as crocin, picrocrocin also exist in other plants, which makes the chemotype-driven analysis not ideal for the quality control of saffron. Herein, we developed a rapid authentication method for saffron in crude drugs by the site-specific PCR. In order to realize fast high-throughput analysis, a one-tube identification approach was further established by using a universal fluorescent dye to detect the PCR products. In addition, this method was also applied to the authentication of saffron in a processed herbal medicine "Er shi wu wei shan hu wan" which consists of twenty-five kinds of medicinal materials including plants, minerals and animals. Additionally, this method was also proved to be with high specificity and repeatability. The flexibility of choosing different primers also made this method versatile for other medicinal materials.

Multiple tests on saffron find new adulterant materials and reveal that Ist grade saffron is rare in the market

Among spices, Saffron is among the most extensively interrogated for purity and authenticity. Numerous methods have been recommended for authentication of Saffron samples and for detection of adulterants for codex compliance. However, none of these methods can fulfill both of these important quality criteria. This study describes a three step approach to achieving this goal by including the established ISO3632 method and two additional methods based on microscopic examination and DNA barcoding. We provide results showing the utility of these methods both independently and in combination for quality evaluation of 36 commercial saffron samples. Our results show that use of the ISO3632 approach alone can reveal the color and aroma but not the genetic origin of the material or distinguish between synthetic components versus natural ingredients. Also, the microscopic observation method can give a preliminary indication of saffron authenticity, but used alone it is unable to quantify purity. Finally, a relatively new method based on the use of DNA barcodes can authenticate the biological origin of the saffron, but here results may be misleading if auto-adulterating materials are present. Overall, our study reveals that through the combined use of all three methods, saffron authentication can substantially improved.

Detection of the most sophisticated saffron fraud with the latest technologies: current fraudulent practices using Gardenia jasminoides extracts

The authenticity of saffron is an important matter in view of consumer protection, quality assurance, active properties and economic impact. Since it is an extremely valuable spice, saffron has been adulterated in various ways. Nowadays there are methods for the assessment of authenticity and detection of adulterants which are included in the quality saffron normative (ISO 3632). Special emphasis is carried out on Gardenia jasminoides fruits adulteration due to its similar chemical composition with saffron.

Quality control of saffron and evaluation of potential adulteration by means of thin layer chromatography-image analysis and chemometrics methods

Saffron (Crocus sativus) stigmas as a flavoring commodity command a very high price in international food markets and, as a result, are a candidate for various kinds of fraud. This paper reports on the use of thin layer chromatography combined with image analysis (TLC-IA) and chemometrics techniques for validating the authenticity of saffron and rapidly identifying the type of adulterants. This method includes several pre-processing steps, such as correcting the general baseline (using the asymmetric least squares (AsLS) algorithm), converting the images to RGB chromatographic channels, and removing the shifts and concavity of spots (using a correlation optimization warping (COW) algorithm) prior to image analysis of saffron thin layer chromatography patterns. After employing the preprocessing sequence, different unsupervised multivariate data analysis (i.e. principal component analysis (PCA) and k-means) and supervised chemometric methods (i.e. partial least squares discrimination analysis (PLS-DA), variable selection (loading weight and variable importance in projection (VIP)) and linear discriminant analysis (LDA)) were applied to validate the authenticity of saffron and to classify the types of adulterants. As a result, quality control of Iranian sourced saffron in the international food market became possible.

Molecular species fingerprinting and quantitative analysis of saffron (Crocus sativus L.) for quality control by MALDI mass spectrometry.

Herein we describe a rapid, simple, and reliable method for the quantitative analysis and molecular species fingerprinting of saffron (Crocus sativus L.) by direct MS and MS/MS analysis. Experimentally, powdered saffron was subjected to a brief treatment with a 0.3% TFA water/acetonitrile solution, and the resulting mixture was directly placed on the MALDI plate for analysis. This approach allowed the detection of the commonly observed crocins C-1–C-6 and flavonols, together with the identification of the unknown highly glycosylated crocins C-7, C-8 and C-9, and carotenoid-derived metabolites. The strategy endorsed the simultaneous detection and characterization of saffron and adulterant markers using crude extracts of the adulterant itself and synthetic sets of adulterated authentic saffron samples. The implementation of the strategy was to measure the amount of an unknown adulterant from the crude extract using curcumin as a non-isotopic isobaric internal standard. The relationship between the saffron and curcumin molar ratios were established with a correlation coefficient of 0.9942. The ANOVA regression model was significant, F(1, 72) = 13 595.82, p < 0.001, y = (0.0116 ± 0.0001)x + (−0.1214 ± 0.0086). No matrix effects were observed and good results were obtained with respect to instrumental repeatability (*RSD% < 2%) and LOD (1.1%). The analysis of commercial samples of saffron using the proposed approach showed the suitability of the method for routine analysis (minimal sample preparation and very short measuring time per sample).

Multivariate statistical methods for the assessment of the quality and the authenticity of saffron

Multivariate statistical methods for the assessment of the quality and the authenticity of saffron

Greek PDO saffron authentication studies using species specific molecular markers

Saffron, the spice produced from the red stigmas of the flower of Crocus sativus L. is a frequent target of fraud and mislabeling practices that cannot be fully traced using the ISO 3632 trade standard specifications and test methods. A molecular approach is proposed herein as a promising branding strategy for the authentication of highly esteemed saffron brands such as the Greek Protected Designation of Origin (PDO) “Krokos Kozanis”. Specific ISSR (inter-simple sequence repeat) markers were used to assess for the first time, the within species variability of several populations of C. sativus L. from the cultivation area of “Krokos Kozanis” as well as the potential differences with the band pattern produced by other Crocus species. Then, species-specific markers were developed taking advantage of an advanced molecular technique such as the HRM analysis coupled with universal DNA barcoding regions (trnL) (Bar-HRM) and applied to saffron admixtures with some of the most common plant adulterants (Calendula officinalis, Carthamus tinctorius, Gardenia jasminoides, Zea mays and Curcuma longa). The sensitivity of the procedure was tested for turmeric as a case study whereas HPLC-fluorescence determination of secondary metabolites was also employed for comparison. The overall results indicated that the Bar-HRM approach is quite effective in terms of specificity and sensitivity. Its effectiveness regarding the detection of turmeric was comparable to that of a conventional HPLC method (0.5% vs 1.0%, w/w). Yet, the proposed DNA-based method is much faster, cost-effective and can be used even by non-geneticists, in any laboratory having access to an HRM-capable real-time PCR instrumentation. It can be, thus, regarded as a strong analytical tool in saffron authentication studies.