Quality evaluation of commercial herbal products using chemical methods.

Herbal products comprise a wide spectrum of locally, nationally or internationally commercialized commodities. As these products have an increasingly important position in healthcare systems worldwide, a detailed product quality assessment is of crucial importance. For the quality evaluation of commercial herbal products, a wide range of methods were used, from simpler, quicker, and cost-effective HPTLC, to hyphenated methods with MS or NMR, where more precise quantification or specific structural information is required. Additionally, most of the methods have been coupled with chemometric tools, such as PCA, or PDA, for the multivariate analysis of the high amount of data generated by chromatograms, electropherograms or spectra. The chemical methods have revealed the widespread presence of low or variable quality herbal products in the marketplace. The majority of analytical investigations present major, qualitative and quantitative, inter-product variations of their chemical composition, ranging from missing ingredients, to strikingly and unnaturally high concentrations of some compounds. Moreover, the inter-batch quality variations were frequently reported, as well as the presence of some undesirable substances. The chemical analysis of herbal products is a vital component to raise the overall awareness of quality in the herbal market and generate a quality driven approach.

Chemical Authentication of Botanical Ingredients: A Review of Commercial Herbal Products.

Chemical methods are the most important and widely used traditional plant identification techniques recommended by national and international pharmacopoeias. We have reviewed the successful use of different chemical methods for the botanical authentication of 2,386 commercial herbal products, sold in 37 countries spread over six continents. The majority of the analyzed products were reported to be authentic (73%) but more than a quarter proved to be adulterated (27%). At a national level, the number of products and the adulteration proportions varied very widely. Yet, the adulteration reported for the four countries, from which more than 100 commercial products were purchased and their botanical ingredients chemically authenticated, was 37% (United Kingdom), 31% (Italy), 27% (United States), and 21% (China). Simple or hyphenated chemical analytical techniques have identified the total absence of labeled botanical ingredients, substitution with closely related or unrelated species, the use of biological filler material, and the hidden presence of regulated, forbidden or allergenic species. Additionally, affecting the safety and efficacy of the commercial herbal products, other low quality aspects were reported: considerable variability of the labeled metabolic profile and/or phytochemical content, significant product-to-product variation of botanical ingredients or even between batches by the same manufacturer, and misleading quality and quantity label claims. Choosing an appropriate chemical technique can be the only possibility for assessing the botanical authenticity of samples which have lost their diagnostic microscopic characteristics or were processed so that DNA cannot be adequately recovered.

The more favorable attitude of the citizens toward GMOs supports a new regulatory framework in the European Union.

Since 1996 till 2018, the global area cultivated with GM crops has increased 113-fold, making biotech crops one of the fastest adopted crop technology in the past decades. In the European Union, only two countries still cultivate one available transgenic crop event on minor hectarage. Moreover, the number of notifications for confined field trials has dramatically dropped in the last decade. All these are happening while the EU legislation on GM crops has come under severe criticism. The percentage of EU citizens concerned about the presence of GMOs in the environment has decreased from 30% (in 2002) to 19% (in 2011), while the level of concern about the use of GM ingredients in food or drinks has decreased from 63% (in 2005) to 27% (in 2019). The steadily increasing acceptance of the EU citizens of GMOs in the environment and food, as it was recorded by Eurobarometers, should additionally ease the way and support a positive change of the legal framework that regulates the GM crops' testing and commercial cultivation in the EU.

Microscopic Authentication of Commercial Herbal Products in the Globalized Market: Potential and Limitations.

Herbal products are marketed and used around the globe for their claimed or expected health benefits, but their increasing demand has resulted in a proportionally increase of their accidental contamination or intentional adulteration, as already confirmed with DNA-based methods. Microscopy is a traditional pharmacopoeial method used for plant identification and we systematically searched for peer-reviewed publications to document its potential and limitations to authenticate herbal medicines and food supplements commercially available on the global market. The overall authenticity of 508 microscopically authenticated herbal products, sold in 13 countries, was 59%, while the rest of 41% were found to be adulterated. This problem was extending over all continents. At the national level, there were conspicuous differences, even between neighboring countries. These microscopically authenticated commercial herbal products confirm that different magnifying instruments can be used to authenticate crude or processed herbal products traded in the global marketplace. The reviewed publications report the successful use of different magnifying instruments, single or in combinations with a second one, with or without a chemical or DNA-based technique. Microscopy is therefore a rapid and cost-efficient method, and can cope with mixtures and impurities. However, it has limited applicability for highly processed samples. Microscopic authentication of commercial herbal products will therefore contribute to raise public awareness for the extent of adulteration and the need to safeguard consumer safety against the challenges of globalization.

Turning Meadow Weeds Into Valuable Species for the Romanian Ethnomedicine While Complying With the Environmentally Friendly Farming Requirements of the European Union's Common Agricultural Policy.

The cross-compliance mechanism of the European Union (EU)'s common agricultural policy (CAP) makes the approval of the direct payments to the European farmers subject to compliance with the requirement to maintain the land in good agricultural and environmental condition. One of the obligations of the Romanian land owners and farmers is to avoid the installation of unwanted vegetation on their land plots. This vegetation is represented by some species of herbaceous or woody plants, annual or perennial, that spontaneously invade the agricultural lands, diminishing the production capacity of the cultivated plants. Included in this category are 10 meadow weeds, without fodder value or even toxic to animals: Arctium lappa L., Carduus nutans L., Conium maculatum L., Eryngium campestre L., Euphorbia cyparissias L., Pteridium aquilinum (L.) Kuhn, Rumex acetosella L., Veratrum album L., Xanthium spinosum L., and Xanthium strumarium L. Various and multiple uses in traditional medicine of these meadow weed species have been reported for Romania and other nine neighboring East European countries, i.e. Bosnia and Herzegovina, Bulgaria, Czech Republic, Estonia, Kosovo, Russia, Turkey, Serbia, and Ukraine. For A. lappa were recorded the highest number of ethnomedicinal uses, in the largest number of East European countries, including Romania. C. maculatum and V. album are not recommended for human consumption but can be further investigated as potential sources of pharmaceutically active compounds. Once removed by landowners and farmers from their land, the raw plant material of these 10 species become readily and easily available to the Romanian local communities and the industry of herbal food supplements, while the biodiversity of the agro-ecosystems is maintained.

A Review of Authenticity and Authentication of Commercial Ginseng Herbal Medicines and Food Supplements.

Ginseng traditional medicines and food supplements are the globally top selling herbal products. Panax ginseng, Panax quinquefolius and Panax notoginseng are the main commercial ginseng species in herbal medicine. Prices of ginseng products vary widely based on the species, quality, and purity of the used ginseng, and this provides a strong driver for intentional adulteration. Our systematic literature search has reviewed the authenticity results of 507 ginseng-containing commercial herbal products sold in 12 countries scattered across six continents. The analysis of the botanical and chemical identity of all these products shows that 76% are authentic while 24% were reported as adulterated. The number of commercial products as well as the percentage of adulteration varies significantly between continents, being highest in South America (100%) and Australia (75%), and lower in Europe (35%), North America (23%), Asia (21%) and Africa (0%). At a national level, from the five countries for which more than 10 products have been successfully authenticated, the highest percentage of adulterated ginseng products were purchased from Taiwan (49%), followed by Italy (37%), China (21%), and USA (12%), while all products bought in South Korea were reported to be authentic. In most cases, labeled Panax species were substituted with other Panax species, but substitution of ginseng root, the medicinally recommended plant part, with leaves, stems or flowers was also reported. Efficient and practical authentication using biomarkers to distinguish the main ginseng varieties and secondary metabolite spectra for age determination are essential to combat adulteration in the global marketplace.

The DNA-Based Authentication of Commercial Herbal Products Reveals Their Globally Widespread Adulteration.

The herbal products, sold worldwide as medicines or foods, are perceived as low risk because they are considered natural and thus safe. The quality of these products is ineffectively regulated and controlled. The growing evidence for their lack of authenticity is causing deep concern, but the scale of this phenomenon at the global, continental or national scale remains unknown. We analyzed data reporting the authenticity, as detected with DNA-based methods, of 5,957 commercial herbal products sold in 37 countries, distributed in all six inhabited continents. Our global survey shows that a substantial proportion (27%) of the herbal products commercialized in the global marketplace is adulterated when their content was tested against their labeled, claimed ingredient species. The adulterated herbal products are distributed across all continents and regions. The proportion of adulterated products varies significantly among continents, being highest in Australia (79%), South America (67%), lower in Europe (47%), North America (33%), Africa (27%) and the lowest in Asia (23%). The commercial HPs' authenticity among the 37 countries included in our global analysis ranges between 0 and 100% from the total number of product reported for each specific national marketplace. For 9 countries, more than 100 products were successfully DNA-based authenticated and reported. From these countries, the highest percentage of adulterated commercial HPs was reported for Brazil (68%), followed distantly by Taiwan (32%), India (31%), USA (29%), followed closely by Malaysia (24%), Japan (23%), South Korea (23%), Thailand (20%), and China (19%). Our results confirm the large-scale presence of adulterated herbal products throughout the global market. The adulterated herbal products contain undeclared contaminant, substitute, and filler species, or none of the labeled species, which all may be accidental or intentional, economically-motivated and fraudulent. Due to the ever-increasing analytical sensitivity of the high throughput DNA sequencing, increasingly used for the untargeted, simultaneous multi-taxa identification, the proportion of adulterated HPs detected on the global market is expected to increase. In the context of the increasing demand for HPs, the limited supply of raw materials derived from many plant species, some of which being already nationally or internationally protected and having various degrees of trade restrictions, adds up to the differences and discrepancies between national HPs' regulatory frameworks and further increases the risks of adulteration of many types of herbal products. The globally widespread adulteration is a serious threat to consumers' well-being and safety, in spite of herbal products' claimed or expected health benefits.

Predation pressure in maize across Europe and in Argentina: an intercontinental comparison.

Humankind draws important benefits from large-scale ecological processes termed ecosystem services, yet the status of several of them is declining. Reliable monitoring methods are essential for tracking the status of ecosystem services. Predation is the mainstay of natural pest control, a key ecosystem service. We used green plasticine caterpillars to monitor predation pressure, and to obtain baseline data on predator activity in transgenic Bt versus non-Bt maize fields in Old and New World countries. Predation pressure was measured at ground and canopy levels using an identical, small-plot experimental design in four European countries (Denmark, Slovakia, Romania and Italy) and Argentina. Total predation rate in maize was 11.7%d-1 (min. 7.2%d-1 in Argentina, max. 29.0%d-1 in Romania). Artificial caterpillars were attacked both by invertebrates (mostly chewing insects with 42.0% of the attack marks, and ants with 7.1%, but also predatory and parasitoid wasps, spiders and slugs), and vertebrates (small mammals 25.5%, and birds 20.2%). Total predation at ground level (15.7%d-1 ) was significantly higher than in maize canopies (6.0%d-1 ) in all countries, except Argentina. We found no significant differences between predator pressure in Bt versus non-Bt maize plots. The artificial caterpillar method provided comparable, quantitative data on predation intensity, and proved to be suitable for monitoring natural pest control. This method usefully expands the existing toolkit by directly measuring ecological function rather than structure.

The Romanian experience and perspective on the commercial cultivation of genetically modified crops in Europe.

Romania was the third country in Europe and the tenth in the world, to commercially adopt genetically modified crops in 1999, only 3 years after they were first marketed globally. Half a million hectares of transgenic herbicide resistant soybean and insect resistant maize were grown in Romania during an uninterrupted 17-year period. After several years of continued declining area, the commercial cultivation of transgenic plants recently ended. The commercial cultivation of transgenic crops in Romania remains legally and technically possible, according to the EU and national regulations. However, the declining area cultivated with these crops in Romania seems to be the result of farmers' conscious decision, while waiting for more profitable genetically modified crops to become available that better fit their needs. This expectation would be a logical result of the EU GMO opt-out Directive 2015/412 and the advent of the new plant breeding techniques. The GMO opt-out mechanism is still expected to unblock the EU authorization process after the large majority of the EU member states have already prohibited the cultivation of genetically modified organisms in their territory. As the new plant breeding techniques offer significant technical and economic advantages, they could be rapidly adopted by commercial breeders and farmers outside Europe. The Court of Justice of the European Union ruled that plants obtained with the new gene editing techniques must go through the same authorization procedure as transgenic plants. This decision is expected to delay the approval and availability of these new plant varieties on the EU market and their commercial cultivation.

What's in the box? Authentication of Echinacea herbal products using DNA metabarcoding and HPTLC.

BACKGROUND: Differences in regulatory policies between countries as well as a lack of appropriate standardized methods for the authentication and quality control of herbal products directly impact their quality and safety. Echinacea products are among the top-selling herbal products in Europe and the United States with indications for a broad range of ailments. The increased use of Echinacea species has led to concerns about adulterated products resulting from challenges in morphology-based identification, due to overlapping morphological variation, frequent hybridization between species, and deliberate adulteration. PURPOSE: This study addressed the need for a novel analytical strategy in the authentication of herbal products. METHODS: A combination of high performance thin layer chromatography (HPTLC) and DNA metabarcoding was employed. Fifty-three Echinacea herbal products marketed across Europe were tested to evaluate the accuracy of these methods in plant identification and their potential for detecting substitutes, adulterants and other unreported plant constituents. RESULTS: HPTLC provides high resolution in the detection of Echinacea phytochemical target compounds, but does not offer information on the other species within the product. Alternatively, we showed that the limitation of HPTLC in detecting non-targeted species can be overcome by the complementary use of DNA metabarcoding. Using DNA metabarcoding, Echinacea species were detected in 34 out of the 38 retained products (89%), but with a lack of discriminatory resolution at the species level due to the low level of molecular divergence within the Echinacea genus. All of the tested herbal products showed considerable discrepancies between ingredients listed on the label and the ones detected using DNA metabarcoding, registering an overall ingredient fidelity of only 43%. CONCLUSION: The results confirm that DNA metabarcoding can be used to test for the presence of Echinacea species and simultaneously to detect other species present in even highly processed and multi-ingredient herbal products.

Benefits and Limitations of DNA Barcoding and Metabarcoding in Herbal Product Authentication.

INTRODUCTION: Herbal medicines play an important role globally in the health care sector and in industrialised countries they are often considered as an alternative to mono-substance medicines. Current quality and authentication assessment methods rely mainly on morphology and analytical phytochemistry-based methods detailed in pharmacopoeias. Herbal products however are often highly processed with numerous ingredients, and even if these analytical methods are accurate for quality control of specific lead or marker compounds, they are of limited suitability for the authentication of biological ingredients. OBJECTIVE: To review the benefits and limitations of DNA barcoding and metabarcoding in complementing current herbal product authentication. METHOD: Recent literature relating to DNA based authentication of medicinal plants, herbal medicines and products are summarised to provide a basic understanding of how DNA barcoding and metabarcoding can be applied to this field. RESULTS: Different methods of quality control and authentication have varying resolution and usefulness along the value chain of these products. DNA barcoding can be used for authenticating products based on single herbal ingredients and DNA metabarcoding for assessment of species diversity in processed products, and both methods should be used in combination with appropriate hyphenated chemical methods for quality control. CONCLUSIONS: DNA barcoding and metabarcoding have potential in the context of quality control of both well and poorly regulated supply systems. Standardisation of protocols for DNA barcoding and DNA sequence-based identification are necessary before DNA-based biological methods can be implemented as routine analytical approaches and approved by the competent authorities for use in regulated procedures. © 2017 The Authors. Phytochemical Analysis Published by John Wiley & Sons Ltd.

Comparative authentication of Hypericum perforatum herbal products using DNA metabarcoding, TLC and HPLC-MS.

Many herbal products have a long history of use, but there are increasing concerns over product efficacy, safety and quality in the wake of recent cases exposing discrepancies between labeling and constituents. When it comes to St. John's wort (Hypericum perforatum L.) herbal products, there is limited oversight, frequent off-label use and insufficient monitoring of adverse drug reactions. In this study, we use amplicon metabarcoding (AMB) to authenticate 78 H. perforatum herbal products and evaluate its ability to detect substitution compared to standard methods using thin-layer chromatography (TLC) and high performance liquid chromatography coupled with mass spectrometry (HPLC-MS). Hypericum perforatum was detected in 68% of the products using AMB. Furthermore, AMB detected incongruence between constituent species and those listed on the label in all products. Neither TLC nor HPLC-MS could be used to unambiguously identify H. perforatum. They are accurate methods for authenticating presence of the target compounds, but have limited efficiency in detecting infrageneric substitution and do not yield any information on other plant ingredients in the products. Random post-marketing AMB of herbal products by regulatory agencies could raise awareness among consumers of substitution and would provide an incentive to manufacturers to increase quality control from raw ingredients to commercialized products.