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Concentrations of mycotoxins determined in individual milk thistle preparations. Numbers above the columns indicate the maximum/minimum concentration value.
Source publication
Herbal-based dietary supplements have become increasingly popular. The extract from milk thistle (Silybum marianum), is often used for the treatment of liver diseases. However, serious concerns exist regarding the efficacy, composition, as well as the safety of these over-the-counter preparations. Therefore, the aim of the present study was to inve...
Contexts in source publication
Context 1
... Generally, the most frequent mycotoxins found in the milk thistle-based preparations were the Fusarium mycotoxins; i.e., type A trichothecenes (HT-2 and T2 toxins occurred in 92% and 96% of the samples, respectively), type B trichothecene deoxynivalenol occurred in 81% of the samples, zearalenone occurred in 89%, cyclic depsipeptidic mycotoxins as enniatines and beauvericin occurring, in 96-100%, and mycotoxins (produced by Alternaria fungi) had an occurrence frequency of 96-100% (Table 2 , Fig. 3). ...
Context 2
... mean concentrations of Fusarium mycotoxins trichothecenes, enniatins, beauvericin, and Alternaria mycotoxins were typically on the order of single digits of mg/kg (for details see Table 2 and Fig. ...
Context 3
... they often dramatically exceeded even the highest maximum limits. For example, the maximum allowed concentration level of HT-2 and T-2 toxins in cereals intended for human consumption is 200 μg/kg 31 ; however, the sum of HT-2 and T-2 toxins in 92% of the investigated milk thistle-based preparations exceeded this value by up to 19× (Table 2 and Fig. ...
Citations
... Positive results have been detected in food supplements containing milk thistle such as dried powder capsules, tablets, teas or seeds. In some cases of significant levels of contamination exceeding the allowed maximum limits according to Commission Regulation (EU) 2023/915 in food-in particular by aflatoxins, T-2 and HT-2 toxins (Tournas et al. 2012;Veprikova et al. 2015;Fenclova et al. 2019;Ałtyn and Twarużek 2020;Pickova et al. 2020;Boško et al. 2022). Metabolites of zearalenone have not been analysed in any study. ...
... Toxic effects can be even more severe when dietary supplements made from multiple plant ingredients with high concentrations of multiple mycotoxins are consumed (Veprikova et al. 2015). Consuming a larger amount could harm the consumer, therefore, it makes sense to collect this data, evaluate it and possibly introduce legislative limits, or regulate the conditions of cultivation and collection of medicinal plants (Steinhoff 2019;Fenclova et al. 2019;Ałtyn and Twarużek 2020). In the European Union, the maximum limits for the content of mycotoxins in plant-based food supplements have not yet been set (Fenclová, 2019). ...
The silymarin complex extracted from milk thistle provides significant health benefits, particularly due to its antioxidant and hepatoprotective properties. However, plant substances can be contaminated by a number of fungi types and their secondary metabolites—mycotoxins. This work deals with the determination of aflatoxins and zearalenone and its metabolites in 39 different samples grown in 2020 and 2021. Analysis of mycotoxins was performed by UHPLC-MS/MS after immunoaffinity column AFLAPREP® and EASI-EXTRACT® ZEARALENONE clean-up. The presence of aflatoxins was not confirmed in the monitored samples, but 1/3 of the samples were contaminated with zearalenone in the range of 2.8–378.9 µg/kg. Metabolites of zearalenone such as α-zearalenol, α-zearalanol, and β-zearalanol were not detected in any of the samples. β-Zearalenol was found in two samples (2.6 µg/kg and 29.8 µg/kg).
... Other main components of silymarin are flavonolignans silychristin (15-25%), silydianin (10%), isosilybin A (10%), 2,3-dehydrosilybin (5%), isosilybin B (<5%) and isosilychristin (3%), along with the flavonoid taxifolin ( Figure 1). 22,23 Silibinin is quite resistant to reduction, but is easily oxidized to 2,3-dehydrosilybin. Long-term heating above 100 °C causes damage to its skeleton structure. ...
Silibinin, a bioactive component found in milk thistle extract (Silybum marianum), is known to have significant therapeutic potential in the treatment of various liver diseases. It is considered a key element of silymarin, which is traditionally used to support liver function. The main mechanisms of action of silibinin are attributed to its antioxidant properties protecting liver cells from damage caused by free radicals. Experimental studies conducted in vitro and in vivo have confirmed its ability to inhibit inflammatory and fibrotic processes, as well as promote the regeneration of damaged liver tissue. Therefore, silibinin represents a promising tool for the treatment of liver diseases. Since the silibinin molecule is insoluble in water and has poor bioavailability in vivo, new perspectives on solving this problem are being sought. The two most promising approaches are the water-soluble derivative silibinin-C-2’,3-dihydrogen succinate, disodium salt, and the silibinin-phosphatidylcholine complex. Both drugs are currently under evaluation in liver disease clinical trials. Nevertheless, the mechanism underlying silibinin biological activity is still elusive and its more detailed understanding would undoubtedly increase its potential in the development of effective therapeutic strategies against liver diseases. This review is focused on the therapeutic potential of silibinin and its derivates, approaches to increase the bioavailability and the benefits in the treatment of liver diseases that have been achieved so far. The review discusses the relevant in vitro and in vivo studies that investigated the protective effects of silibinin in various forms of liver damage.
... 9 Without chemical standardization of the botanical being tested, dosages of the active compounds are unknown, results can vary from batch to batch, and preclinical as well as clinical studies cannot be reproduced. 10,11 Variations in the composition of silybin from milk thistle results from different growth conditions 12,13 and variations in extract preparation and handling. 13−15 Therefore, a validated analytical method for the measurement and standardization of silymarin constituents in test materials is essential for the support of pharmacological and toxicological studies both in vitro and in vivo and to enable the correlation of activities with dosage. ...
Extracts prepared from the seeds of the medicinal plant milk thistle [Silybum marianum (L.) Gaertn. (Asteraceae)] are widely used as dietary supplements due to anti-inflammatory, antitumor, and hepatoprotective effects. Called silymarin, the main components of lipophilic extracts of milk thistle seeds are flavonoids and flavonolignans including silybin A, silybin B, isosilybin A, isosilybin B, silydianin, silychristin, taxifolin, and 2,3-dehydrosilybins. The aim of this study was to develop a method based on UHPLC-MS/MS for the chemical authentication and standardization of milk thistle silymarin. Validation included the method of standard addition to account for the lack of a blank matrix. Potential matrix effects were investigated by analyzing silymarin standards dissolved only in the initial UHPLC mobile phase. Measurements of six flavonolignans and taxifolin in the milk thistle extract using UHPLC-MS/MS with standard addition or external standard calibration produced similar results for all analytes except silydianin and 2,3-dehydrosilybin B, which showed significant peak enhancement during negative ion electrospray due to botanical matrix effects. The UHPLC-MS/MS-based method of standard addition requires <10 min per injection and is suitable for the standardization of silymarin from milk thistle in support of preclinical and clinical studies of safety and efficacy.
... Two of the milk thistle products were from the United States, and the other two from the Czech Republic. 20 The increased sale of highly diluted or "empty" extracts also has been noted at BotaniCERT, a contract analytical laboratory based in Grasse, France. A review of test results obtained by UHPLC with ultraviolet/visible detection (UHPLC-UV/Vis) over the period from 2017 to 2019conducted on 379 crude (whole, cut, or powdered) botanical ingredients and 1,028 extracts -showed no crude materials lacking the expected constituents, but 11% of the extracts were considered "empty" or devoid of any constituents characteristic for the labeled or any other botanical ingredient ( Figure 3). ...
CELEBRATING 40 YEARS This article discusses the composition and labeling of botanical extracts, with particular focus on the sale of botanical dietary ingredients and dietary and food supplement products. Published literature and reports from analysts suggest that some commercial extracts in the dietary and food supplement supply chain may contain little or none of the declared plant or plant extract. Experts in dietary supplement analysis have determined that this occurs mainly because of two schemes used by deceptive suppliers. First, certain suppliers excessively dilute native plant extracts with undeclared amounts of excipients and are ambiguous in disclosing the plant-to-extract ratio. Second, whole herbs are extracted to obtain specific fractions or compounds that are considered to be therapeutically beneficial and are provided to select markets. The marc (leftover or spent biomass) may then be resold without disclosure that it is pre-extracted material.
... Two of the milk thistle products were from the United States, and the other two from the Czech Republic. 20 The increased sale of highly diluted or "empty" extracts also has been noted at BotaniCERT, a contract analytical laboratory based in Grasse, France. A review of test results obtained by UHPLC with ultraviolet/visible detection (UHPLC-UV/Vis) over the period from 2017 to 2019conducted on 379 crude (whole, cut, or powdered) botanical ingredients and 1,028 extracts -showed no crude materials lacking the expected constituents, but 11% of the extracts were considered "empty" or devoid of any constituents characteristic for the labeled or any other botanical ingredient ( Figure 3). ...
This article discusses the composition and labeling of botanical extracts, with particular focus on the sale of botanical dietary ingredients and dietary and food supplement products. Published literature and reports from analysts suggest that some commercial extracts in the dietary and food supplement supply chain may contain little or none of the declared plant or plant extract. Experts in dietary supplement analysis have determined that this occurs mainly because of two schemes used by deceptive suppliers. First, certain suppliers excessively dilute native plant extracts with undeclared amounts of excipients and are ambiguous in disclosing the plant-to-extract ratio. Second, whole herbs are extracted to obtain specific fractions or compounds that are considered to be therapeutically beneficial and are provided to select markets. The marc (leftover or spent biomass) may then be re-sold without disclosure that it is pre-extracted material.
... These contaminants can potentially cause liver damage with the use of herbal-based dietary supplements. [120,121] Phyllanthus niruri (Chanca piedra/Gale-ofthe-wind/Bhumi amla) ...
Complementary and alternative medicines (CAM) include conventional medical treatments. Patients worldwide use CAM at alarming rates; thus, reports of CAM-related DILI have been on the rise. The clinical presentations include asymptomatic liver test abnormalities, acute hepatitis with or without jaundice, acute cholestatic liver disease (bland or with hepatitis), acute liver failure, severe hepatitis with features of portal hypertension, and acute decompensation of known or unknown cirrhosis that can lead to acute-on-chronic liver failure. Acute hepatitis with or without necrosis, hepatocellular and canalicular cholestasis, herb-induced or CAM-triggered autoimmune hepatitis, granulomatous hepatitis, severe steatohepatitis, and vanishing bile duct syndrome are common liver biopsy findings in CAM-DILI. The presence of preexisting liver disease predicts severe liver injury, risk of progression to liver failure, and decreased transplant-free survival in patients with CAM-DILI. This review discusses global epidemiology and trends in CAM-DILI, clinical presentation, assessment and outcomes, commonly emerging threats in the context of hepatotoxic herbs, pragmatic assessment of “liver beneficial” herbs and health care myths, patient communication, regulatory framework, and future directions on research in CAM.
... Despite the therapeutic benefits of silymarin, its weak biochemical properties (a lack of solubility in water) and limited bioavailability caused unsatisfactory and inconsistent therapeutic results, which leads to more instances of drug-drug interactions when other concurrent medications are given [27,28] which led to search for innovative method to solve such problems as using nanotechnology. Nanotechnology offers the chance to complement natural remedies by facilitating the identification, manufacture, and implementation of a wide range of therapeutic strategies to improve health and lessen the severity of various diseases [25]. ...
Background
Cholestasis is an important predisposing factor for hepatocyte damage, liver fibrosis, primary biliary cirrhosis, and even liver failure. Silybum marianum L. (SM) plant is used in teas or eaten in some countries due to its antioxidant and hepatoprotective properties. Because of its low and poor oral bioavailability, so we improve the therapeutic activity of Silybum marianum L. extract (SM) by studying the potential effects of nanoformulation of Silybum marianium L. extract (nano-SM) on 17α-ethinylestradiol (EE)-induced intrahepatic cholestasis.
Methods
Thirty female Sprague-Dawley rats were divided into 5 groups (6 rats/group). Group I: Rats were received the treatment vehicle and served as normal group. Group II:Rats were injected daily with EE (10 mg/kg) for five successive days. Group III-V: Rats were injected daily with EE (10 mg/kg) and treated with either Ursodeoxycholic acid (UDCA) (40 mg/kg), SM (100 mg/kg) and nano-SM (100 mg/kg) orally once/day throughout the trialfor five successive days, respectively.
Results
Nano-SM greatly dampened the increase in serum levels of total and direct bilirubin, alanine aminotransaminase, aspartate aminotransaminase, and alkaline phosphatase caused by EE. Furthermore, nano-SM increased the hepatic contents of reduced glutathione (GSH) and catalase (CAT) and also upregulated the relative hepatic gene expressions of Rho-kinase (ROCK-1), myosin light chain kinase (MLCK), and myosin phosphatase target subunit (MYPT1) compared to the EE-induced group. Administration of nano-SM reduced hepatic lipid peroxidation and downregulated the relative hepatic expressions of the nuclear factor-kappa B (NF-ҡB) and interleukin-1β (IL-1β). In addition, nano-SM improved the histopathological changes induced by EE.
Conclusion
Nano-SM possessed a superior effect over SM, which can be considered an effective protective modality against EE-induced cholestatic liver injury through its antioxidant, anti-inflammatory activities, and enhancing bile acid (BA) efflux.
... Nevertheless, the chemical composition of the milk thistle used material was rarely determined in most of the studies focusing on its biological activity [52]. Thus, notwithstanding the purported beneficial role of milk thistle, the vague description of the chemical composition of the derived preparations represents probably, for several studies, the main pitfall in proving its clinical efficacy [52,55,56]. Moreover, widespread contamination with fungi, microbes, and pesticides has been reported in milk thistle-based dietary supplements raising serious safety issues for human health, as botanicals-induced hepatotoxicity may occur [55,57,58]. ...
... Thus, notwithstanding the purported beneficial role of milk thistle, the vague description of the chemical composition of the derived preparations represents probably, for several studies, the main pitfall in proving its clinical efficacy [52,55,56]. Moreover, widespread contamination with fungi, microbes, and pesticides has been reported in milk thistle-based dietary supplements raising serious safety issues for human health, as botanicals-induced hepatotoxicity may occur [55,57,58]. Studies have shown alarming discrepancies between declared and detected chemical content between brands of marketed milk thistle preparations, as well as within batches of the same preparations and manufacturers. ...
... Studies have shown alarming discrepancies between declared and detected chemical content between brands of marketed milk thistle preparations, as well as within batches of the same preparations and manufacturers. These differences can critically alter the expected therapeutic effects [51,55,56,59,60]. Some of the studies focusing on the quantitative analysis of silymarin showed that a large number of investigated marketed preparations contained a lower amount than declared and some were even completely missing this marker compound of milk thistle [51,53,59,61]. ...
Background
Milk thistle is one of the most popular hepatoprotectants, and is often sold in combination with other ingredients. Botanical supplements are known to be vulnerable to contamination and adulteration, and emerging technologies show promise to improve their quality control.
Methods
Untargeted and semi-targeted metabolomics based on UHPLC-QTOF-ESI⁺MS techniques, UV spectrometry, and DNA metabarcoding using Illumina MiSeq were used to authenticate eighteen milk thistle botanical formulations (teas, capsules, tablets, emulsion).
Results
Untargeted metabolomics separated 217 molecules and by multivariate analysis the discrimination between the different preparations was established. The semi-targeted metabolomics focused on 63 phytochemicals, mainly silymarin flavonolignans and flavonoids, that may be considered as putative biomarkers of authenticity. All formulations contained molecules from silymarin complexes at different levels. The quantitative evaluation of silybins was done using in parallel UV spectrometry and UHPLC-QTOF-ESI⁺MS and their correlations were compared. DNA metabarcoding detected milk thistle in eleven out of sixteen retained preparations, whereas two others had incomplete evidence of milk thistle despite metabolomics validating specific metabolites, e.g., silymarin complex, identified and quantified in all samples. Meanwhile, the DNA metabarcoding provided insights into the total species composition allowing the interpretation of the results in a broad context.
Conclusion
Our study emphasizes that combining spectroscopic, chromatographic, and genetic techniques bring complementary information to guarantee the quality of the botanical formulations.
... MT food supplements were found to have alarming discrepancies between declared and detected chemical content on individual preparations and within batches of the same preparations and manufacturers [19,20,[24][25][26]. Most of the investigated preparations contained a significantly lower amount of silymarin than declared on the label of the product, some completely missing silymarin, therefore critically altering the expected therapeutic effects [19,20,[24][25][26]. Notwithstanding, MT-derived preparations have been recognized as being safe for human consumption [3,6,27], their widespread contamination with fungi, microbes, and pesticides has been widely reported raising alarming safety MT-based food supplements are available in a variety of pharmaceutical formulations, including pills, tablets, capsules, or standardized extracts, and can be bought at pharmacies and over largely uncontrolled distribution channels, such as e-commerce platforms [3,18,19]. ...
... With attracting claims on their purported health benefits, food supplements are used by consumers with the hope of maintaining their health or supporting specific health conditions. Studies show that up to 43% of patients with liver disease and up to 87% of patients with cancer use hepatoprotective herbal-based food supplements, including MT [20]. The high popularity is spurred, among others, by the consumers' misconception that botanicals are "natural" and, thus, always "safe" [21]. ...
... Inadequate regulatory frameworks and poor quality control strategies are enhancing economically motivated fraud, and a large number of adulterated MT products that include depleted extracts and synthetic colorants were detected in the global market [20,22,23]. MT food supplements were found to have alarming discrepancies between declared and detected chemical content on individual preparations and within batches of the same preparations and manufacturers [19,20,[24][25][26]. ...
Milk thistle (Silybum marianum (L.) Gaertn.) is among the top-selling botanicals used as a supportive treatment for liver diseases. Silymarin, a mixture of unique flavonolignan metabolites, is the main bioactive component of milk thistle. The biological activities of silymarin have been well described in the literature, and its use is considered safe and well-tolerated in appropriate doses. However, commercial preparations do not always contain the recommended concentrations of silymarin, failing to provide the expected therapeutic effect. While the poor quality of raw material may explain the low concentrations of silymarin, its deliberate removal is suspected to be an adulteration. Toxic contaminants and foreign matters were also detected in milk thistle preparations, raising serious health concerns. Standard methods for determination of silymarin components include thin-layer chromatography (TLC), high-performance thin-layer chromatography (HPTLC), and high-performance liquid chromatography (HPLC) with various detectors, but nuclear magnetic resonance (NMR) and ultra-high-performance liquid chromatography (UHPLC) have also been applied. This review surveys the extraction techniques of main milk thistle metabolites and the quality, efficacy, and safety of the derived food supplements. Advanced analytical authentication approaches are discussed with a focus on DNA barcoding and metabarcoding to complement orthogonal chemical characterization and fingerprinting of herbal products.
... Dietary supplements may be composed of multiple components, but if only one component is of poor quality, it still can reduce the quality of the final product [29][30][31][32][33]. Results can vary widely, and in some cases plant raw material contamination with aflatoxins never exceeded the LOD [34]. In milk thistle [35], the most common mycotoxins were HT-2 (92%), T2 (96%), DON (81%) and ZEN (89%). We found similar levels of ZEN contamination in the present study of purple coneflower extracts. ...
Purple coneflower (Echinacea purpurea (L.) Moench) is a plant in the family Asteraceae, mainly grown in North America. Echinacea purpurea has been used in conventional medicine. The plant has immuno-stimulating and antibacterial properties, but neither mold contamination nor a mycotoxin presence have been evaluated. Our goal is to determine the degree to which molds and mycotoxins contaminate dietary supplements based on purple coneflower distributed on the Polish market. We analyzed 21 samples divided into four groups: sachets (n = 5), dry raw material (n = 3), capsules (n = 9), and tablets (n = 4). The mycological analysis of dietary supplements shows that the average number of molds is 1012 cfu/g, and the most common molds are Aspergillus spp., Phoma spp. and Eurotium spp. The mycotoxins most common in the samples are ZEN (18/21), DON (5/21) and T-2 toxin (3/21).
