madman
Super Moderator
Abstract
Objective: Nutraceutical products are widely used for their claimed therapeutic benefits. However, falsified or adulterated nutraceuticals present a major health threat to consumers. This study investigates the pharmaceutical quality, safety, and anti-inflammatory effects of six male enhancement nutraceuticals that claim to be 100% natural.
Methods: Three batches of six male enhancement products were tested to detect the presence and levels of adulterants via high-performance liquid chromatography (HPLC). The pharmaceutical quality of the selected nutraceuticals was tested with near-infrared spectroscopy (NIR) and SeDeM. The cytotoxic effects of these products on HepG2 cells were determined through cell proliferation (XTT) and lactate dehydrogenase (LDH) cytotoxicity assays. Lastly, the in vitro inflammatory effects of these products were investigated using murine J774 macrophages through cytokine release analysis.
Results: HPLC analysis detected the presence of sildenafil citrate, a vasodilator, and the active ingredient in Viagra and Revatio, in all batches of the products we analyzed. The amount of sildenafil citrate ranged from 0.45 mg to 51.85 mg among different batches. NIR assessment showed inter-and intra-batch heterogeneity in product composition.
Results of the XTT and LDH assays showed significant cytotoxic effects of the analyzed products. XTT analysis revealed that the viability of HepG2 treated with tested products varied from 27.57% to 41.43%. Interestingly, the male enhancement products also showed anti-inflammatory effects.
Conclusion: Despite their labeling as 100% natural, all products tested in this study contained levels of sildenafil citrate, which was not reported on the packaging. There was a lack of pharmaceutical uniformity among products of the same batch and across different batches. Additionally, the products we tested had cytotoxic effects. These study findings highlight the adulteration, poor quality, and hazard of these nutraceuticals. Therefore, strict regulation of these products and standardization of the definition of nutraceuticals are urgently needed. Further, these falsely advertised products should be withdrawn from the market due to potential adverse effects on the health of their consumers.
1. Introduction
The term nutraceutical has been defined as ‘‘food or part of a food that provides medical or health benefits, including the prevention and/or treatment of a disease” [1]. However, about 25 definitions of nutraceutical exist in the scientific literature; they can be contradictory and no single definition is well accepted worldwide. Most of these definitions associate nutraceuticals with food, food components, or nutrients that can provide health benefits [2]. Nutraceutical products that claim to have therapeutic benefits for erectile dysfunction (ED), obesity, and cognitive impairment are increasingly advertised and sold [3]. Nutraceutical ingredients, such as vitamins, carotenoids, bioactive peptides, polyunsaturated fatty acids, and plant polyphenols, are considered promising approaches to alleviate certain skin conditions and to delay premature skin aging [4]. However, most of these claims lack support from human trials or other reliable scientific research.
Several nutraceuticals have been advertised to help symptoms of ED [1]. Online retail companies, such as Amazon, eBay, and Alibaba, are the main sellers of these types of products in the United States [3]. ED is the failure to attain or maintain a sufficient erection for satisfactory sexual performance [5]. It has been suggested that over 80% of ED cases have an organic etiology (i.e., endocrine or non-endocrine etiologies), and the remaining cases are due to psychogenic disorders [5]. According to Yafi et al. [5], the initial management of ED is life-style modification, followed by first-line treatment with phosphodiesterase 5 inhibitors (PDE-5Is) and vacuum erection devices. In addition, people seek male enhancement nutraceuticals with the misconception that they are 100% natural and thus possess few or no side effects. However, many of these products were found to contain undeclared compounds that are PDE-5Is [6].
The reliability of nutraceutical products is complicated by the absence of strict regulatory laws that govern claims made on their labels [1]. The claimed beneficial health effects of nutraceuticals are mainly unsupported by appropriate in vivo research; rather, they are based on in vitro studies. Also, in vivo studies are required for a better understanding of the possible side effects, the mechanism(s) of action, and bioavailability of nutraceutical products [7].
It has been reported that many nutraceutical products are falsified or adulterated. Hence, in addition to not having the advertised properties, they may represent a major health threat to consumers [1,6]. It was reported that the three top-selling nutraceuticals in the Egyptian market for ED claimed to be 100% natural, but they contained undeclared sildenafil citrate, an active pharmaceutical ingredient (API) for managing ED, which can have serious side effects and contraindications [6]. Another study reported the adulteration of weight-loss nutraceutical products with various APIs, including sibutramine, sildenafil citrate, orlistat, and phenolphthalein, as well as poor pharmaceutical and microbial qualities [8]. Many of the nutraceutical products sold online in the U.S. were found to be adulterated with undeclared APIs [1]. This evidence suggests that vendors exploit the hopes of people who are looking to treat a condition and market products that make false claims about their ingredients, safety, and possible side effects.
Attempts by the Food and Drug Administration (FDA) to regulate nutraceuticals were met with public and legislative resistance, as demonstrated by the adoption of the Vitamins and Minerals Act of 1976 and the Dietary Supplement and Health Education Act of 1994, both of which restricted the FDA’s ability to remove products that were misbranded or posed raised safety concerns based on clinical evidence [9]. In 2006, due to the high number of products appearing on the market, the European Commission passed a regulation to ensure that consumers were not misled by claims about the healing properties of nutraceutical [10]. In Japan, nutraceuticals fall under the generic term ‘‘health foods” that encompasses many categories, such as nutraceuticals, health supplements, natural foods, and dietary supplements [11]. There is a need for a universally applicable definition and for laws that regulate the manufacturing and label claims of nutraceutical products [1,12]. Additionally, the scientific community and regulatory agencies are responsible for drawing attention to the possible deleterious effects of these products. For example, the FDA has issued a warning letter to companies promoting products claiming to prevent or treat COVID-19 [13]. Also, the European Medicines Agency has released a warning that asks the public not to buy medicines from unauthorized websites or vendors who claim that their products can cure or prevent COVID-19 because these products are likely to be counterfeit [14]. The limited control of the FDA over dietary supplements and the absence of regulatory laws that govern various aspects of the nutraceutical industry are the main reasons for such misleading claims [1,3].
This study investigated whether nutraceuticals that advertised male enhancement effects were 100% natural, as their labels claimed, and also evaluated their safety and pharmaceutical quality. These analyses were conducted for AlphaMan XL, MACA, Prime Test, Golden X, Horny Goat Weed, and Leyzene, six of the most-used products in this field. High-performance liquid chromatography (HPLC) was used to detect the presence of adulterants such as sildenafil citrate, tadalafil, and vardenafil and to quantify their abundance if present in these products. Near-infrared spectroscopy (NIR) was used to test the pharmaceutical quality of the selected nutraceutical products. Possible cytotoxic effects of these products were investigated using cell proliferation kit II (XTT) and lactate dehydrogenase (LDH) cytotoxicity assay kit. Lastly, the inflammatory or anti-inflammatory effects of the tested products were investigated.
5. Conclusion
The male enhancement nutraceuticals that we studied were proven to be spiked with an undeclared adulterant, sildenafil citrate, which can be particularly harmful to individuals with hypertensive and cardiac disorders. Additionally, the tested products suffered from poor pharmaceutical quality and had high inter-and intra-batch heterogeneity, as confirmed through HPLC, NIR and SeDeM analyses. The safety of the male enhancement nutraceuticals we analyzed was investigated by measuring the integrity of HepG2 membrane using the LDH assay and detecting the mitochondrial activity with the XTT assay. Both LDH and XTT assays showed that the product samples had high toxicity towards HepG2 cells. Finally, the male enhancement products we studied also showed anti-inflammatory effects.
In conclusion, these kinds of products require careful regulation by local regulatory affairs agencies, such as FDA in the U.S., the Department of Health in Canada and the Canadian Food Inspection Agency in Canada.
Objective: Nutraceutical products are widely used for their claimed therapeutic benefits. However, falsified or adulterated nutraceuticals present a major health threat to consumers. This study investigates the pharmaceutical quality, safety, and anti-inflammatory effects of six male enhancement nutraceuticals that claim to be 100% natural.
Methods: Three batches of six male enhancement products were tested to detect the presence and levels of adulterants via high-performance liquid chromatography (HPLC). The pharmaceutical quality of the selected nutraceuticals was tested with near-infrared spectroscopy (NIR) and SeDeM. The cytotoxic effects of these products on HepG2 cells were determined through cell proliferation (XTT) and lactate dehydrogenase (LDH) cytotoxicity assays. Lastly, the in vitro inflammatory effects of these products were investigated using murine J774 macrophages through cytokine release analysis.
Results: HPLC analysis detected the presence of sildenafil citrate, a vasodilator, and the active ingredient in Viagra and Revatio, in all batches of the products we analyzed. The amount of sildenafil citrate ranged from 0.45 mg to 51.85 mg among different batches. NIR assessment showed inter-and intra-batch heterogeneity in product composition.
Results of the XTT and LDH assays showed significant cytotoxic effects of the analyzed products. XTT analysis revealed that the viability of HepG2 treated with tested products varied from 27.57% to 41.43%. Interestingly, the male enhancement products also showed anti-inflammatory effects.
Conclusion: Despite their labeling as 100% natural, all products tested in this study contained levels of sildenafil citrate, which was not reported on the packaging. There was a lack of pharmaceutical uniformity among products of the same batch and across different batches. Additionally, the products we tested had cytotoxic effects. These study findings highlight the adulteration, poor quality, and hazard of these nutraceuticals. Therefore, strict regulation of these products and standardization of the definition of nutraceuticals are urgently needed. Further, these falsely advertised products should be withdrawn from the market due to potential adverse effects on the health of their consumers.
1. Introduction
The term nutraceutical has been defined as ‘‘food or part of a food that provides medical or health benefits, including the prevention and/or treatment of a disease” [1]. However, about 25 definitions of nutraceutical exist in the scientific literature; they can be contradictory and no single definition is well accepted worldwide. Most of these definitions associate nutraceuticals with food, food components, or nutrients that can provide health benefits [2]. Nutraceutical products that claim to have therapeutic benefits for erectile dysfunction (ED), obesity, and cognitive impairment are increasingly advertised and sold [3]. Nutraceutical ingredients, such as vitamins, carotenoids, bioactive peptides, polyunsaturated fatty acids, and plant polyphenols, are considered promising approaches to alleviate certain skin conditions and to delay premature skin aging [4]. However, most of these claims lack support from human trials or other reliable scientific research.
Several nutraceuticals have been advertised to help symptoms of ED [1]. Online retail companies, such as Amazon, eBay, and Alibaba, are the main sellers of these types of products in the United States [3]. ED is the failure to attain or maintain a sufficient erection for satisfactory sexual performance [5]. It has been suggested that over 80% of ED cases have an organic etiology (i.e., endocrine or non-endocrine etiologies), and the remaining cases are due to psychogenic disorders [5]. According to Yafi et al. [5], the initial management of ED is life-style modification, followed by first-line treatment with phosphodiesterase 5 inhibitors (PDE-5Is) and vacuum erection devices. In addition, people seek male enhancement nutraceuticals with the misconception that they are 100% natural and thus possess few or no side effects. However, many of these products were found to contain undeclared compounds that are PDE-5Is [6].
The reliability of nutraceutical products is complicated by the absence of strict regulatory laws that govern claims made on their labels [1]. The claimed beneficial health effects of nutraceuticals are mainly unsupported by appropriate in vivo research; rather, they are based on in vitro studies. Also, in vivo studies are required for a better understanding of the possible side effects, the mechanism(s) of action, and bioavailability of nutraceutical products [7].
It has been reported that many nutraceutical products are falsified or adulterated. Hence, in addition to not having the advertised properties, they may represent a major health threat to consumers [1,6]. It was reported that the three top-selling nutraceuticals in the Egyptian market for ED claimed to be 100% natural, but they contained undeclared sildenafil citrate, an active pharmaceutical ingredient (API) for managing ED, which can have serious side effects and contraindications [6]. Another study reported the adulteration of weight-loss nutraceutical products with various APIs, including sibutramine, sildenafil citrate, orlistat, and phenolphthalein, as well as poor pharmaceutical and microbial qualities [8]. Many of the nutraceutical products sold online in the U.S. were found to be adulterated with undeclared APIs [1]. This evidence suggests that vendors exploit the hopes of people who are looking to treat a condition and market products that make false claims about their ingredients, safety, and possible side effects.
Attempts by the Food and Drug Administration (FDA) to regulate nutraceuticals were met with public and legislative resistance, as demonstrated by the adoption of the Vitamins and Minerals Act of 1976 and the Dietary Supplement and Health Education Act of 1994, both of which restricted the FDA’s ability to remove products that were misbranded or posed raised safety concerns based on clinical evidence [9]. In 2006, due to the high number of products appearing on the market, the European Commission passed a regulation to ensure that consumers were not misled by claims about the healing properties of nutraceutical [10]. In Japan, nutraceuticals fall under the generic term ‘‘health foods” that encompasses many categories, such as nutraceuticals, health supplements, natural foods, and dietary supplements [11]. There is a need for a universally applicable definition and for laws that regulate the manufacturing and label claims of nutraceutical products [1,12]. Additionally, the scientific community and regulatory agencies are responsible for drawing attention to the possible deleterious effects of these products. For example, the FDA has issued a warning letter to companies promoting products claiming to prevent or treat COVID-19 [13]. Also, the European Medicines Agency has released a warning that asks the public not to buy medicines from unauthorized websites or vendors who claim that their products can cure or prevent COVID-19 because these products are likely to be counterfeit [14]. The limited control of the FDA over dietary supplements and the absence of regulatory laws that govern various aspects of the nutraceutical industry are the main reasons for such misleading claims [1,3].
This study investigated whether nutraceuticals that advertised male enhancement effects were 100% natural, as their labels claimed, and also evaluated their safety and pharmaceutical quality. These analyses were conducted for AlphaMan XL, MACA, Prime Test, Golden X, Horny Goat Weed, and Leyzene, six of the most-used products in this field. High-performance liquid chromatography (HPLC) was used to detect the presence of adulterants such as sildenafil citrate, tadalafil, and vardenafil and to quantify their abundance if present in these products. Near-infrared spectroscopy (NIR) was used to test the pharmaceutical quality of the selected nutraceutical products. Possible cytotoxic effects of these products were investigated using cell proliferation kit II (XTT) and lactate dehydrogenase (LDH) cytotoxicity assay kit. Lastly, the inflammatory or anti-inflammatory effects of the tested products were investigated.
5. Conclusion
The male enhancement nutraceuticals that we studied were proven to be spiked with an undeclared adulterant, sildenafil citrate, which can be particularly harmful to individuals with hypertensive and cardiac disorders. Additionally, the tested products suffered from poor pharmaceutical quality and had high inter-and intra-batch heterogeneity, as confirmed through HPLC, NIR and SeDeM analyses. The safety of the male enhancement nutraceuticals we analyzed was investigated by measuring the integrity of HepG2 membrane using the LDH assay and detecting the mitochondrial activity with the XTT assay. Both LDH and XTT assays showed that the product samples had high toxicity towards HepG2 cells. Finally, the male enhancement products we studied also showed anti-inflammatory effects.
In conclusion, these kinds of products require careful regulation by local regulatory affairs agencies, such as FDA in the U.S., the Department of Health in Canada and the Canadian Food Inspection Agency in Canada.
